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Thermo Scientific HyClone Single-Use Bioreactor (SUB) User`s Guide

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1. n j 75 13 71 55 a 1908 2 47 05 41 05 1195 0 40 17 1042 6 1020 4 y l y y 819 2 35 45 900 4 37 88 E 962 0 HyClone Laboratories Inc Logan UT USA is ISO Certified 2010 Thermo Fisher Scientific Inc All rights reserved All other trademarks are the property of Thermo Fisher Scientific Inc and its subsidiaries Specifications terms and pricing are subject to change Not all products are available in all countries Please consult your local sales representative for details Cell Culture amp BioProcessing 925 West 1800 South In Americas Asia In Europe www thermoscientific com hyclone Logan UT 435 792 8000 32 53 85 71 80 84321 435 792 8001 fax 32 53 85 74 31 fax ha Gan a a a a a O Section 3 9 Load Cells Kit Data Sheet 3 9 Load Cells Kit 39 1 Load Cells Kit Data Sheet Data Sheet Thermo Scientific Single Use Bioreactor S U B 178 Load Cell kits are available for the Thermo Scientific HyClone Single Use Bioreactor S U B and Single Use Mixer S U M Kits can be configured for new or existing units D mh D N J D D mh co gt D lt N Thermo Scientific HyClone Load Cell Kits Overview The FLEXMOUNT Weigh Modules are be load cell equipped as an option for the used to enable weight measurements They 50 500 L U B and the 50 2000 L S U M T operate as a mechanical suspension system units Load cells come standard on the on the Thermo Scie
2. _______ p Part of Thermo Fisher Scientific SCIENTIFIC Data Sheet Ordering Instructions Standard 48 series vent filter heaters for use with Pall KA3 vent filters selecting a complete kit will require two part numbers a vent filter heater controller SV50191 01 for 115 VAC or SV50191 02 for 230 VAC and the suitable power cord for your installation SV50191 02 or 04 through 08 NOTE SV50191 10 does not have a controller and is for custom installations only Part Number Description Additional Information SV50191 07 Power Cord 240 VAC For use in United Kingdom BS1363 with 12 leads and GFCI SV50191 08 Power Cord 240 VAC For use in Europe CEE 7 7 with 12 leads and GFCI SV50191 09 Extension Cord for Series 48 Heater 100 240 VAC For use between heater and controller 10 Vent Filter Heater with RTD two wire Pt 100 pee 9V50191 10 100 120 VAC 17 24 W Includes sheathed 20 pig tail lead NEMA rated 48 series vent filter heater kits for use with Pall KA3 vent filters sold as a kit which includes vent filter heater controller with a water tight enclosure quick connects and installation power cord SV50191 06 Power Cord 100 120 VAC For use in U S Japan NEMA 5 15P with 12 leads and GFCI Part Number Description Additional Information S V50191 11 NEMA rated Vent Heater w Programmable Controller Includes low temp alarm preset temp 50 C 100 120VAC power cord power cord w flying l
3. Reusable tubing clamp used on probe ports when connecting probe assemblies SV50177 01 Autoclave Tray for Probe Kits stainless steel ae Eh Vie eNO E pa DSC AE tale AGOE EE Available for aseptic luer connection or weld onto the sample line to take sample sets Vent Filter Heater and Cord Options k Probe Bioreactor Probe Autoclave Tray A a Elevated Dual Assembly for Probe Kits S Handie Filter Brackets Es y and Filter Heater ts Threaded Probe i pu w7 Attachment n C cable Tie gt i Bioreactor Probe SYA A Molded Bellows Autoclave Tray Frit Sparge Line ul a Support i Additional information on autoclave tray e Fabricated from stainless steel _ Cable Tie e Plastic carry handle for easy transport right out of the autoclave A e Positions probes on 15 incline for ter probe membrane longevit AE Pall ACD Connector PEIEE ply DENY a e Will restrain probe bellows from a collapsing during sterilization e Probe holder accommodates two probes SV50223 2000 L SH30774 07 or SH30774 08 SH30720 01 SV20664 03 SV50177 01 SV20750 01 SH30845 02 Bioreactor control system S U B Hardware System standard S U B BPC standard with or without condenser bag assembly Bioreactor Probe Assembly non sterile for use in autoclave Heavy Duty Tubing Clamp 10 pack Autoclave Tray for Autoclaving Probe Kits S U B Temperature Port used to verify RTD calibration Sterile Sampling Manifold with L
4. Thermo Scientific Single Use Bioreactor S U B 198 Section 5 4 Ordering Information Partial Spare Parts List Partial Spare SSS Parts List Cont Drive Shaft Assembly 2000 L S U B Four 4 Piece Integrated Design V50177 160 Motor 1 4HP 208VAC 3Ph IP65 10 1 Gear Reduction 50 L 100 L S U B s SV50177 161 Motor 1 4HP 208VAC 3Ph IP65 12 5 1 Gear Reduction 250 L S U B SV50177 162 Motor 1 2HP 208VAC 3Ph IP65 15 1 Gear Reduction 500 L 1000 L S U B s RTD Standard PT100 3 Wire 60 With Quick Disconnect Bulgin 3 pin Connector SVOOT7 234 pater to P N TX 10289 Cable Assy Disposable Sensor Fiber Optic DO Sensor Green Body Uo Mates to SV20963 sensors SV50177 193 Lamp Green 120VAC Replacement for motor on off switch 9V50177 194 Lamp Green 240VAC Replacement for motor on off switch Cable Assy Disposable Sensor Fiber Optic PH Sensor Blue Body uaa Mates to SV20963 sensors 5V50177 238 Pressure Transducer Disposable 12 length QC w 4 Wire pigtail Thermo Scientific Single Use Bioreactor S U B 199 5 5 Ordering Instructions 5 6 Ordering Support Contact Information Technical Support Technical Service Hotline Initial Setup and Operation Training Thermo Scientific Section 5 5 Ordering Information Replacement amp Spare Parts Disposable and hardware components for the Single Use Bioreactor S U B can be ordered directly from your sales representat
5. 2000 L S U B Systems Include 304 stainless steel outer support container variable speed agitation controller motor drive assembly with four segment drive shaft RTD sensor integrated water jacket three load cells with summing block display BPC pressure monitor probe support system and standard tool set Part Number Description SV50223 11 2000 L S U B Water Jacket Modbus Plus Interface 240 VAC SV50223 12 2000 L S U B Water Jacket Allen Bradley Interface 240 VAC SV50223 13 2000 L S U B Water Jacket Analog Output Interface 240 VAC SV50223 14 2000 L S U B Water Jacket Profibus Interface 240 VAC SV50223 15 2000 L S U B Water Jacket DeviceNet Interface 240 VAC Thermo Scientific Single Use Bioreactor S U B 195 Section 5 2 Ordering Information Hardware The S U B Condenser System consists of a cart and brackets a condenser chill plate a peristaltic pump and a chiller temperature control unit or TCU Part Number Description Complete Condenser System 120 V including cart chill plate and 9V50232 01 nae mounting post with filter brackets chiller and pump Complete Condenser System 240 V including cart chill plate and 9V50232 02 ae mounting post with filter brackets chiller and pump SV50232 71 Condenser assembly including chill plate and mounting post with filter brackets Option Allowing two chill plates per system SV50232 23 Thermoflex 900 TCU 115 VAC 60 Hz with necessary plumb
6. 30 cm and 3 8 9 5 mm ID X 5 8 15 9 mm OD C Flex X 12 30 cm Bottom Drain Harvest Hydrophobic vent filter with Emflon II 1 4 6 4 mm ID X 7 16 11 1 mm OD C Flex X 6 15 cm reduced to 1 8 3 2 mm ID X 1 4 6 4 mm OD C Flex X 60 152 cm I9 3 8 9 5 mm ID X 5 8 15 9 mm OD C Flex X 60 152 cm splits to ee Tene minty 1 4 6 4 mm ID X 7 16 11 1 mm OD C Flex X 12 30 cm reduced to ialecittan allie zseb 1 8 3 2 mm ID X 1 4 6 4 mm OD C Flex X 12 30 cm and 3 8 MPC Bod 3 8 9 5 mm ID X 5 8 15 9 mm OD C Flex X 12 30 cm y Pall Kleenpak aseptic 1 2 12 7 mm ID X 3 4 19 1 mm OD C Flex X 6 15 cm splits to connector KPCHT Series 1 2 12 7 mm ID X 3 4 19 1 mm OD C Flex X 6 15 cm and Female 1 2 12 7 mm ID X 3 4 19 1 mm OD C Flex X 10 25 cm Pall Kleenpak 0 2 Micron Exhaust Vent Filter a 1 4 6 4 mm ID X 7 16 11 1 mm OD C Flex X 60 152 cm reduced to ae t AON 1 3 3 2 mm ID X 1 4 6 4 mm OD C Flex X 12 30 cm apee 1 4 6 4 mm ID X 7 16 11 1 mm OD C Flex X 3 8 cm reduced to Check Valve and 1 4 6 4 mm ID X 7 16 11 1 mm OD C Flex X 48 122 cm with Emflon Il Porous frit gas sparge 1 4 6 4 mm ID X 7 16 11 1 mm OD C Flex X 6 15 cm reduced to 12 mm dia 25 um pores Check Valve and 1 4 6 4 mm ID X 7 16 11 1 mm OD C Flex X 48 122 cm with Emflon Il Custom BPC Options
7. C Flex platinum cured silicone PVC Pharmed PharmaPure More information in selection guide of BPC catalog specific lengths of 1 8 ID to 1 ID D limitations due to port size Luers CPC Quick Connects SIP connectors Triclamp Kleenpak Lynx Refer to selection guide in BPC catalog for further SmartSite Clave Lynx steam thru CPC Steam Thru Gore steam information by connection type Note Only option valve Gore Mini TC BioQuate GE PAW SterilEnz end plug etc for probe port connections is Kleenpak Additional ports second row of 4 for 50 L to 250 L S U Bs second row of 5 for 500 L and 1000 L S U B s Pressure sensor Pendotech for 50 L to 250 L S U B comes standard on 500 L and 1000 L DO Finesse DO and pH PreSens Availability of various other disposable sensors currently pending the hn doa Limited engineer to order customization possible such as additional Dependent on location in bag and compatibility media lines and vent filter lines Requires economic justification with hardware Dependent on location in bag and fit with hardware Limited customization possible as engineer to order with justification Fea 1 ID porton harvest ine Limited customization possible e g moving sample tempwell port to a probe tube port or swapping overlay inlet line with supplement line Dual sparger open pipe plus porous frit standard Can do an engineer to order for one or the other alone Make to
8. 8V50177 38 Drive Shaft Assembly 1000 L S U B Three 3 Piece Integrated Design Delrin SST Drive Shaft Assembly 1000 L S U B Four 4 Piece Integrated Design Delrin SST SV50177 39 ae NON Released Engineering Design SV50177 40 Drive Shaft Assembly 250 L S U B One 1 Piece Integrated Design Delrin SST 9V50177 41 Drive Shaft Assembly 250 L S U B Two 2 Piece Integrated Design Delrin SST 9V50177 65 Frit Sparge Line Support SV50177 90 Safety Cap Drive Shaft Motor Assembly Clear Lexan SV50177 110 Cable Assembly 16 Pin Main communication cable from S U B S U M to External Controller Male one end SV50177 112 Cable Assembly 16 Pin Main communication cable from S U B S U M to External Controller Male 2 Male SV50177 113 Cable Assembly 16 Pin Main communication cable from S U B S U M to External l Controller Male one end VFD amp Temperature control only SV50177 114 Cable Assembly 3 Pin Pressure Alarm Relay cable from S U B S U M to External Controller Male 2 Male SV50177 117 Cable Assembly 4 Pin TCU Communicaiton cable Male one end Cable Assembly 4 Pin Disposable pressure sensor communication cable w Buccaneer 9V50177 118 QC Male amp Female components SV50177 119 Cable Assembly 16 Pin w 3 Pin RTD in line QC pigtail Main communication cable from S U B S U M to External Controller Male one end amp 3 Pin OC pigtail 9V50177 124 1 4 Dia Neoprene Rubber Cord
9. Section 1 2 Hardware Characteristics Heating times for the S U B systems vary based upon operating liquid volume and temperature ambient or heating fluid temperature sparge rate and mixing rate Users should adjust process liquid staging and seeding strategies to the unique aspects of the S U B Process controllers and heaters in resistive heating systems are designed to provide optimum heat transfer and to minimize heat up times while maintaining the material integrity of the polymer film construction of the S U B BPC Refer to Section 1 8 for expected heating times Users can choose to bypass the temperature and mixing speed control system provided with the S U B and utilize existing bioreactor controllers Refer to Section 1 8 for more details Load cells are standard on the 1000 and 2000 L S U B hardware and optional for the smaller S U B volumes Load cell kits can be installed at the factory or can be added later by a certified service technician The load cell kit comes with three load cells summing block wiring and display with a choice of several data interfaces Load cells arrive uncalibrated it is recommended that the load cell manufacturer or a qualified technician calibrate these systems on site Single Use Bioreactor S U B 6 Section 1 3 End User Supplied Components 1 3 End User Supplied Components pH and Dissolved The following table shows the length and diameter requirements for Oxyg en DO Probes t
10. Thermo Scientific Single Use Bioreactor S U B 5 Section 2 3 single Use Bioreactor BPC Loading 1000 L Electric Resistive Heater 6 Remove the bubble wrap from the sparge filters Guide the sparge inlet lines and filters through the bottom cutouts in the outer support container Figure 2 39 the operator can reach just below the tank to further extend the sparge lines from the cutouts Figure 2 40 Figure 2 39 7 Figure 2 40 Pass the bagged drain line set and temperature sampling port set through the large cut out in the front of the outer support container Figure 2 41 Extend the drain line set through the cut out Figure 2 42 bs Figure 2 41 Figure 2 42 Thermo Scientific Single Use Bioreactor S U B 52 Section 2 3 single Use Bioreactor BPC Loading 1000 L Electric Resistive Heater Gas Supply The BPC should be partially inflated to allow for proper insertion of the Connections and Air drive shaft and to aid in proper alignment of the BPC in the outer support container Inflation 8 Connect pressure transducer to monitor and once the display has stabilized tare the monitor the monitor should be allowed to warm up for 30 minutes before taring Verify that the monitor reads zero 9 Attach air supply to overlay gas inlet line NOTE Air pressure to the overlay gas line on the S U B BPC should not exceed 3 psi less than 25 lpm a WARNING DO NOT EXCEED 0 5 psi 0 34 bar within the BPC or the syste
11. l Section 2 4 Single Use Bioreactor BPC Loading 2000 L Water Jacket The access doors should be secured first with the latches Proper tension is obtained by adjusting the threaded latch pin Tension of the latch is adjusted by varying the position of the pin on the threaded shank The proper latch tension can be obtained by a combination of feel and visual inspection When closing the latch the handle should begin to provide resistance to closing when the leading edge of the safety pin pass through of the latch handle aligns with the outside edge of the latch base Figure 2 136 NOTE When the latch is under tensioned the safety pin pass through of the latch handle will be covered within the latch base and the handle will close very easily If the latch is over tensioned the handle will be excessively difficult to close A For maximum security insert pins not included into the respective latches Turn off the air supply to the overlay line Insert drive shaft and close side access doors Outside Edge of Latch Base Safety Pin Pass Through of Latch set Latch Retainer Safety Pin Pass Through of Latch Handle Figure 2 136 1 Verify the proper position of the exhaust filters and that the exhaust flow path is unobstructed Connect the gas supply lines verify the intended flow paths for overlay frit sparges and open pipe sparge Verify the overlay and direct sparge lines are well positioned and free of kin
12. middle section with internal external threads on each end and lower section with the square end Lubricate the threaded ends with a light coat of food grade anti seize with each use 18 First insert the lower section through the hollow pass through of the mixer drive Figure 2 51 Slide latch pin from mixing assembly into shaft to prevent it from falling into the tube Assemble the middle and lower sections of the drive shaft by joining the segments with a twisting motion fasten the two sections together Figure 2 52 NOTE Segmented shaft threads are left handed Locate one wrench on the flat area in the middle drive shaft section another wrench on the lower section and tighten the connection using a counterclockwise rotation Figure 2 53 The shafts are reverse threaded to avoid loosening during operation Once the sections are secure return the wrenches to the tool holder CAUTION Do not over tighten a snug fit is sufficient Remove the latch pin Figure 2 51 Figure 2 52 Figure 2 53 Thermo Scientific Single Use Bioreactor S U B 56 Thermo Scientific Section 2 3 single Use Bioreactor BPC Loading 1000 L Electric Resistive Heater Load the partially assembled drive shaft through the hollow pass through and hold it in position sufficient to allow a wrench to be placed on the upper most flats of this shaft section Obtain the upper section of the drive shaft and assemble it to the other segment in the manner descri
13. Y jouju04 dwaj sauiejuoy Woddns HEIER Single Use Bioreactor S U B 135 Section 3 3 4 250 L Control Panel Layout 3 3 4 250 L Control Panel Layout 250 L Control Panel Electric Resistive Heater Main Power Source 16 Pin Communication Quick Connect speed Control Key Pad Motor Controller On Off Switch Temperature Controller External Heater j Relay Input TH gt an Emergency Stop Heater Cable Main Switch zT Buzzer i Motor Cable 250 L Control Panel Water Jacket Speed Control Key Pad Main Power Source m 16 Pin Communication Quick Connect Motor Controller On Off Switch Emergency Stop Main Switch Buzzer gt Motor Cable Thermo Scientific Single Use Bioreactor S U B 136 3 4 500 L Single Use Thermo Scientific Bioreactor 3 4 1 3 4 2 3 4 3 3 4 4 Section 3 4 500 L Single Use Bioreactor 500 L S U B Data Sheet 500 L Technical Specifications Electric Resistive Heater 500 L Technical Specifications Water Jacket 500 L Control Panel Layout Single Use Bioreactor S U B 137 The Thermo Scientific HyClone Single Use Bioreactor S U B is the leading single use alternative to conventional stirred tank bioreactors for animal and insect cell culture The current range includes units with maximum working volumes of 50 100 250 500 1000 and 2000 L The S U B is now available with new optimized standard B
14. flow rate of gas into S U B DO probes require routine maintenance replace damaged probe or membrane Verify DO probe calibration relative to set points of zero and span Inspect line sets connected to direct sparges for restriction closed tubing clamp pinched line low supply pressure Direct membrane sparge does not seem to be working although gas is present Allow sparge membrane to purge If the S U B is filled with liquid and allowed to sit idle for extended periods of time without gas being supplied to the sparge liquid can accumulate between the membrane and check valve Various media additives may restrict the membrane temporarily Several minutes of gas pressure being supplied to the sparge should purge the membrane allowing it to function properly Certain operating conditions can create situations when the sparge membrane may become restricted due to insufficient line pressure from the bioreactor controller gas feed line Increasing the flow rate to 1 liter per minute or momentarily raising the pressure regulator outlet pressure to 5 psi 0 34 bar may alleviate the problem Alternatively several seconds at this higher pressure will allow the membrane to purge pores that may be blocked due to the presence of accumulated liquid Single Use Bioreactor S U B 187 Thermo Scientific Symptom Remedy Symptom Remedy Symptom Remedy Section 4 Troubleshooting DO readings are erratic or unstable Ad
15. 19 1 mm ID X 1 25 4 mm OD C Flex X 48 122 cm splits to Harvest 1 4 6 4 mm ID X 3 8 9 5 mm OD C Flex X 12 30 cm and Pall Kleenpak Male 1 2 12 7 mm ID X 3 4 19 1 mm OD C Flex X 24 61 cm 1 2 12 7 mm ID X 3 4 19 1 mm OD C Flex X 10 25 cm and injection site assembly 1 2 12 7 mm ID X 3 4 19 1 mm OD C Flex X 10 25 cm and 3 8 MPC Bod m 1 2 12 7 mm ID X 3 4 19 1 mm OD C Flex X 3 8 cm reduced to Pe Recon 1 4 6 4 mm ID X 7 16 11 1 mm OD C Flex X 6 15 cm reduced to Plugged Feed Line 1 2 12 7 mm ID X 3 4 19 1 mm OD C Flex X 3 8 cm reduced to Base Addition 1 4 6 4 mm ID X 7 16 11 1 mm OD C Flex X 84 213 cm reduced to Plugged 1 8 3 2 mm ID X 1 4 6 4 mm OD C Flex X 12 30 cm Condenser Bag Assembly 14 Condensate Return Line Condenser Bag Assembly Condenser Bag Assembly Condenser Bag Assembly Pi te SALE 1 4 6 4 mm ID X 7 16 11 1 mm OD C Flex X 6 15 cm converge to one Kleenpak Emflon Il Capsule nun 1 4 6 4 mm ID X 7 16 11 1 mm OD C Flex X 72 183 cm 1 4 6 4 mm ID X 7 16 11 1 mm OD C Flex X 72 183 cm reduced to Kleenpak Eon Il Capsule Open Pipe Gas Sparge Check Valve and 1 4 6 4 mm ID X 7 16 11 1 mm OD C Flex X 72 183 cm Sleenpak Emtion Il Capsule Bottom Drain 1 2 12 7 mm ID X 3 4 19 1 mm OD C Flex X 24 61 cm reduced to 1 4 MPC Insert and 1 2 12 7 mm ID X 3 4 19 1 mm OD C Flex X
16. Figure 2 75 opening the safety cover and removing the threaded cap turn counterclockwise Figure 2 76 of the mixing assembly Figures 2 77 ry Figure 2 75 Figure 2 76 Cap gt l PA Drive Shaft Head Hollow b Motor Drive Keyway Pass Through die 3 Safety Cover Drive Shaft gt Figure 2 77 Mixing Assembly Thermo Scientific Single Use Bioreactor S U B 67 Thermo Scientific 16 17 Section 2 3 Single Use Bioreactor BPC Loading 1000 L Water Jacket Verify the three sections of the drive shaft are located in the drive shaft holders on the side of the outer support container The sections will be referred to as the upper section with the drive shaft head middle section with internal external threads on each end and lower section with the square end Lubricate the threaded ends with a light coat of food grade anti seize with each use First insert the lower section through the hollow pass through of the mixer drive Figure 2 78 Once inserted slide latch pin from mixing assembly into shaft to prevent it from falling into the tube Figure 2 79 Assemble the middle and lower sections of the drive shaft by joining the segments and with a twisting motion fasten the two sections together Figure 2 80 NOTE Segmented shaft threads are left handed Locate one wrench on the flat area in the middle drive shaft section another wrench on the lower section and tighten the connection
17. Heater Breakers 3 Pressure Sensor Display Load Cell Motor Controller Switch 3 way Temperature Controller Switch 2 way Temperature Controll y iaa ae a apt TET AN y S S By 5 ail Heater Breakers 3 ih palo j ae Ei no heater non operational nS mae or E Pressure Sensor Display Load Cell Motor Controller Switch 3 way Temperature Controller Switch 2 way Figure 1 11 Control Box for 1000 L U B Jacketed Thermo Scientific Single Use Bioreactor S U B 18 Load Cell Preparation Thermo Scientific gt FIXED PIN TOP PLATE JAM NUT 7 Section 1 6 Initial Installation Instructions For S U B hardware units purchased with factory installed load cells the load cells are shipped in the locked position centering washer and nut are threaded up for equipment protection Figure 1 12 and 1 13 A 15 16 open box wrench is recommended for unlocking the load cells To unlock the load cells loosen the centering washer and nut 5 8 nut until the nut is threaded down on the hold down bolt about half way Next loosen the jam nut on the base plate until the jam nut and centering washer nut meet Tighten both nuts together Using an open end wrench loosen the hold down bolt by moving it upward to provide clearance below and above the cutout in the top plate Figure 1 65 Section A A The clearance around above and below this nut should be fr
18. The S U B BPC is not rated as a pressure vessel All gas supplied to the bioreactor controller must be regulated to a pressure not to exceed manufacturer s recommendations typically 10 psi 0 69 bar Gas pressure in the S U B BPC headspace should not exceed 0 5 psi 0 03 bar at any time e More demanding applications may warrant an optional exhaust vent heater See Section 5 3 e If foaming is excessive in your cell culture process it is best to reduce the operating volume of the process to 80 of maximum rated working volume of the S U B system being used to provide greater head space volume e Disposable pressure transducers are available on custom S U B configurations and are standard on the 1000 L and 2000L systems This technology combined with high level control systems common with industrial applications can regulate gas pressure within the confines of the S U B Extensive cell culture testing has not found an occurrence of overpressure sufficient to create a containment breach Development testing of the S U B BPC system has shown that in conditions of excessive pressure the polymer bag will fail at the upper regions of the chamber where it is unsupported by the outer support container minimizing the likelihood of the loss of bulk liquid Single Use Bioreactor S U B 26 Dispense and Harvest Agitation Hardware Operating Information Temperature Electric Resistive Heating Thermo Scientific Sys
19. The S U B is not equipped with level sensors However the S U B can be setup to allow supplement feeds and volumes to be managed by weight Please refer to Section 3 for skid dimensions for sizing of scales Single Use Bioreactor S U B 188 Section 4 Troubleshooting Symptom pH levels are questionable or out of range Remedy Verify calibration of probe utilize either media or gas buffers e pH levels can be managed in a similar manner to conventional bioreactors once calibration of the probe is verified by use of an off line sample Carbon dioxide gas sparged through the media or headspace biocarbonate levels in the media and the addition of liquid titrant solutions all serve to manage the pH balance of the bioreactor environment See Section 2 5 Probe Assembly instructions for more information on probe calibration e A grounding reference to the media can be created by using a grounding lead between the tank and the body of the stainless steel DO probe or to the stainless steel connector if present on the sample line of the S U B BPC Symptom S U B BPC seems overly tight or overpressure Remedy Verify bag is venting and inspect for cause of overpressure e Reduce inlet gas flow rate of overlay and direct sparge Inspect exhaust filter for restriction or blockage e Excessive foaming should be avoided for several reasons If foam levels are allowed to reach the exhaust filter the filter will become restricted resu
20. a simple process based reference for determining If a vent heater option will be sufficient or if a Condenser system is preferred Typically any off gassing flow rates approaching 50 Ipm or culture durations exceeding 10 days should consider utilizing the Condenser System A l D N N 9 T 28 35 42 Run Duration days Condenser System 2 X 10 vents Vent Heater System 2 X 10 vents Vent Heater System 2 X 3 vents Vent Heater System 1 X 3 vents Graph 1 Condenster vs Exhaust Vent Filter Heaters 1 Exhaust Line From S U B A 11 Filter Bracket Assembly 10 Filter Straps d _ 12 Condenser Post Assembly r i m 17 Condenser Plate _ Assembly 2 Condenser Return Line Back to S U B 16 Dual Headed Peristaltic Pump 9 Exhaust Vent Filters 13 Post Receivers 8 Condenser Bag Gas Outlet Port 3 Condenser Bag Gas Inlet Port 14 Cart 7 Dual Chamber Assembly Condenser Bag 15 Closed 4 Gripping Bath Tabs Recirculating Chiller 6 Condenser Bag 5 Alignment Holes Liquid Drain Ports he f Condenser Disposables Condenser Hardware Condenser System Figure 1 Condenser System Overview Table 2 Condenser System Overview Data Sheet Part Number Description 2300 584 2
21. and Single Use Bioreactor S U B 3 Thermo Scientific Section 1 1 S U B Overview portable dolly The drive shaft is detachable and reusable and is inserted into the BPC through the mixing assembly and into the bearing port Load cells are offered standard on the 1000 and 2000 L S U B s and are optional for the smaller volume systems The BPC includes the impeller assembly sparger vent filter inlet outlet ports probe integration ports and filling dispensing and sampling ports Each S U B BPC comes fully assembled and gamma irradiated The materials are fully qualified for biological product contact per USP Class VI plastics Each assembly is manufactured under cGMP and is supported by qualification and validation information No reuse cleaning or sterilization validations are required as the BPC is provided gamma irradiated Innovative proprietary technology allows for the integration of the mixing shaft pH and dissolved oxygen probes and the resistive temperature detector RTD The probe and temperature interfaces are comparable to traditional systems with the design allowing for simple aseptic connections Integrated disposable spargers are built into the S U B BPC through universal ports The control system encompasses both incorporated and end user supplied components Controls for temperature resistive only and agitation are integrated into the S U B with pH dissolved oxygen DO probes and controls being supp
22. guidelines below to setup the Single Use Bioreactor upon arrival Warning Any procedures that require the control box to be open should be performed with the main electrical disconnect in the locked out position and all power sources removed from the control box For operator safety secure the location of the S U B outer support container by disabling the swivel casters before servicing A 1 S U B units are shipped with the electrical breakers in the OFF position to prevent accidental activation of the heaters during hardware installation NOTE The heater should only be enabled when a liquid filled BPC is present Using a flat head screwdriver open the control box and locate the breakers for the PID temperature controller middle of top row and heater located in middle of lower row Figure 1 6 These breakers should be in the upper ON position during operation For electrical schematics please refer to the ETP which is located on CD provided with hardware 2 Verify position of the three way motor controller switch it should be in the middle position For reference the middle position is to enable the speed control keypad the top position is for 0 10V controllers and the bottom is for 4 20mA controllers Verify position of the two position temperature control toggle switch it should be in the up position for reference the upper position enables the PID temperature controller Single Use Bioreactor S U B 14 Section 1 6
23. possible to the body of the S U B BPC A 7 Important During media fill verify the position of all critical ports drain spargers line sets and probes before more than 50 L of liquid is filled into the BPC This will still allow time for adjustments if it is required after the fill is initiated 8 Typically two each fill lines 2 x 34 and peristaltic pumps are recommended to fill the 2000 L S U B in a timely manner Thermo Scientific Single Use Bioreactor S U B 92 Section 2 4 single Use Bioreactor BPC Loading 2000 L Water Jacket Figure 2 140 Figure 2 141 Probe Insertion 1 Probe clips for the 2000 L S U B do not require a probe tray They 2000 L clip onto the outer support container and hang from above the probe assembly Figure 2 142 2 The probe clip attaches to the tank and is retained using an adjustable ball detent Figure 2 142 3 Install presterilized sensor and probe kit using aseptic connection methods as described in Section 2 7 4 The aseptic connection is completed prior to the bellows being collapsed Figure 2 143 Figure 2 142 Figure 2 143 Thermo Scientific Single Use Bioreactor S U B 93 Thermo Scientific Section 2 4 single Use Bioreactor BPC Loading 2000 L Water Jacket Insert the probe by collapsing the bellows Figure 2 144 NOTE If the BPC is already filled with liquid the best practice is to squeeze the bellows to expel air prior to collapsing it N
24. 13 mm X 13 5 PG thread w 195 235 mm insertion length Bioreactor control system DO probe 1 1 Peristaltic Pump Used for fluid transfer between line sets and containers Temperature Control Unit TCU Necessary for water jacket temperature control not provided by Thermo Scientific a u i re Hardware Features 100 L with Resistive Heater The stainless steel outer support container contains the following features 1 Mixer motor 6 Drive shaft Ze Exhaust vent filter holder 7 Liquid sight window 3 Stainless steel 304 outer support 8 Standard tool set container J Probe access window 4 Bearing port receiver with clamp 10 Control panel D Mixing assembly with shield 11 Probe holder Probe shelf Platform with swivel casters and leveling pads Resistive heater w heat shield Bottom cut outs for BPC alignment Stretch hooks Hardware Features 100 L with Water Jacket The stainless steel outer support container contains the following features 1 Mixer motor 7 Liquid sight window Z Exhaust vent filter holder 8 Standard tool set 3 Stainless steel 304 outer support 9 Probe access window container 10 Control panel 4 Bearing port receiver with clamp 11 Probe holder 5 Mixing assembly with shield AD Probe shelf 6 Drive shaft 14 15 16 17 Platform with swivel casters and leveling pads Laser Dimple Water Jacket Bottom cut outs for BPC alignment Stretch hooks Jacket quick
25. 3 4 19 1 mm OD C Flex X 6 15 cm splits to connector KPCHT Series 12 Exhaust Line 1 2 12 7 mm ID X 3 4 19 1 mm OD C Flex X 6 15 cm and Female 1 2 12 7 mm ID X 3 4 19 1 mm OD C Flex X 10 25 cm Pall Kleenpak 0 2 Micron Exhaust Vent Filter AEE E EN Snare 1 4 6 4 mm ID X 7 16 11 1 mm OD C Flex X 3 8 cm reduced to Hydrophobic vent filter PANEN parg Check Valve and 1 4 6 4 mm ID X 7 16 11 1 mm OD C Flex X 48 122 cm with Emflon II Porous frit gas sparge 1 4 6 4 mm ID X 7 16 11 1 mm OD C Flex X 6 15 cm reduced to Hydrophobic vent filter 12 mm dia 25 um pores Check Valve and 1 4 6 4 mm ID X 7 16 11 1 mm OD C Flex X 48 122 cm with Emflon Il Custom BPC Options C Flex platinum cured silicone PVC Pharmed PharmaPure More information in selection guide of BPC catalog Specific lengths of 1 8 ID to 1 ID ID limitations due to port size Luers CPC Quick Connects SIP connectors Triclamp Kleenpak Lynx Refer to selection guide in BPC catalog for further SmartSite Clave Lynx steam thru CPC Steam Thru Gore steam information by connection type Note Only option valve Gore Mini TC BioQuate GE PAW SterilEnz end for probe port connections is Kleenpak Erect et a row of 5 for 500 L and 1000 L S U B s Pressure sensor Pendotech for 50 L to 250 L S U B comes standard on 500 L and 1000 L DO Finesse DO and pH PreSens Av
26. 4 10 cm splits to SterilEnz Pouch with 3 4 19 1 mm ID X 1 25 4 mm OD C Flex X 84 213 cm splits to pail cicsanak aseni Media Fill 1 2 12 7 mm ID X 3 4 19 1 mm OD C Flex X 24 61 cm and Sn n 1 2 12 7 mm ID X 3 4 19 1 mm OD C Flex X 24 61 cm 1 8 3 2 mm ID X 1 4 6 4 mm OD C Flex X 84 213 cm 12 E Eee Stane 1 2 12 7 mm ID X 3 4 19 1 mm OD C Flex X 4 10 cm reduced to Kleenpak Emflon II Capsule y parg 3 8 9 5 mm ID X 5 8 15 9 mm OD C Flex X 84 213 cm and Pressure transducer 3x 12 mm PDVF Porous Sparge Inserts connected to 3 Tri Clamp Pot PNA Gasket end cap and clamp OD Custom BPC Options C Flex platinum cured silicone PVC Pharmed PharmaPure More information in selection guide of BPC catalog specific lengths of 1 8 ID to 1 ID ID limitations due to port size Luers CPC Quick Connects SIP connectors Triclamp Kleenpak Lynx Refer to selection guide in BPC catalog for further SmartSite Clave Lynx steam thru CPC Steam Thru Gore steam information by connection type Note Only option valve Gore Mini TC BioQuate GE PAW SterilEnz end plug etc for probe port connections is Kleenpak Additional ports second row of 4 for 50 L to 250 L S U Bs second row of 5 for 500 L and 1000 L U B s Pressure sensor Pendotech for 50 L to 500 L S U B comes standard on 1000 L and 2000 L DO pH Pressure Finesse DO and pH
27. Access Doors The procedures listed below must be followed in full detail and in proper Resistive Heater Only order to ensure the S U B outer support container is operated in a safe manner The outer support container is subjected to high structural loads due to the weight and pressure of the fluid and gas contained in the BPC In order to ensure structural integrity of the system it is necessary for the tension on each door be even and involves a two step tensioning procedure of the latches Neglecting these instructions may result in personal injury and or permanent damage to hardware system The access doors of the outer support container should never be opened when the BPC is filled with liquid 24 Close both the left and right side access doors of the outer support container while paying close attention to ensure the BPC is not pinched in any of the door seams Thermo Scientific Single Use Bioreactor S U B 58 Section 2 3 single Use Bioreactor BPC Loading 1000 L Electric Resistive Heater 25 Each door is supplied with four side latches the closure sequence and tension method must be performed in the following manner Figure 2 58 Following this sequence will ensure that the outer support container will retain the proper shape and structure after repeated use Figure 2 58 Latch Closure Sequences latch references are shown in circles 26 The doors should be secured first with the two middle latches latches 2 and 3 Pr
28. Close clamp nearest purge bag 8 Open clamp to sample bag 50 mL pillow bag 9 Allow bag to fill with liquid by gravity recommended 10 20 mL 10 Close clamps at sample manifold inlet ee a 11 Close clamp nearest the sample bag 12 Remove the filled manifold from S U B by welding new manifold onto sample line this will be used for taking the next sample Figure 2 156 Figure 2 157 Thermo Scientific Single Use Bioreactor S U B 99 Section 2 7 Pall Kleenpak Connector Instructions 2 71 Pall Kleenpak Connector Instru ctions These instructions and the information contained within must be read thoroughly as they contain valuable information gained by extensive experience It is very important that all instructions are carefully followed and where appropriate they should be incorporated into the end user s standard operating procedures If some of the procedures do not suit your needs please consult Pall or your local distributor before using Kleenpak connectors Use of this product in a manner other than in accordance with Pall s current recommendations may lead to injury or loss Pall cannot accept liability for such injury or loss Installation and Operation Training on the proper assembly and recommended uses of the Kleenpak connector is necessary e Before installation it is essential to verify that the Kleenpak connector selected is suitable for the fluid to be processed and has been sterilized appropriately e Co
29. Figure 1 9 Control Box for 50 100 250 and 500 L Water Jacket S U B For resistive systems The S U B units are shipped with the electrical breakers in the OFF position to prevent accidental activation of the heaters during hardware installation NOTE The heater should only be enabled when a liquid filled BPC is present Using a flat head screwdriver open the control box and locate the breakers for the Proportional Integral Derivative PID temperature controller heaters pressure sensor and load cells middle of top row Figure 1 10 These breakers should be in the upper ON position during operation For electrical schematics please refer to the ETP which is located on CD provided with hardware NOTE The heater and PID breakers are present in the Water Jacket versions but non operational Verify position of the three way motor controller switch it should be in the middle position for reference the middle position is to enable the speed control keypad For resistive systems Verify position of the two position temperature control toggle switch it should be in the up position for reference the upper position enables the PID temperature controller NOTE This switch is present in the Water Jacket versions but non operational Close the control box and lock the panel using a screwdriver before continuing Single Use Bioreactor S U B 17 Section 1 6 Initial Installation Instructions Temperature Controller Breaker
30. Initial Installation Instructions 3 Close the control box and lock the panel using a screwdriver before continuing 4 To unlock the load cells refer to Load Cell Preparation instructions below Temperature Controller Breaker Heater Breaker wired in 50 100 and 500 L only Heater Breaker wired in 250 and 500 L only Motor Controller Switch 3 way Temperature Controller Switch 2 way _ pa Figure 1 6 Control Box for 50 100 250 and 500 L Electric Resistive Heater S U B Load Cell Preparation 5 For S U B hardware units purchased with factory installed load cells the load cells are shipped in the locked position centering washer and nut are threaded up for equipment protection Figure 1 7 and 1 8 A 15 16 open box wrench is recommended for unlocking the load cells To unlock the load cells loosen the centering washer and nut 5 8 nut until the nut is threaded down on the hold down bolt about half way Next loosen the jam nut on the base plate until the jam nut and centering washer nut meet Tighten both nuts together Using an open end wrench loosen the hold down bolt by moving it upward to provide clearance below and above the cutout in the top plate Figure 1 61 Section A A The clearance around above and below this nut should be from 1 8 to 3 16 Once the proper clearance has been achieved retighten the jam nut Thermo Scientific Single Use Bioreactor S U
31. Installation of Female l Electrical Receptacle In order complete this installation the facility must be equipped with an electrical housing of sufficient size e Typically in the U S the plug will require a two gang box when using the adapter plate supplied e For installations outside the U S an adapter plate is not supplied it is recommended that an electrical panel be modified to accommodate the cutout dimensions as shown below 106mm gt WALT 4 5 5mm DIA Figure 1 Panel Cutout 2 Thermo Scientific Verify that electrical power has been disconnect and been locked out for safety Verify the holes for mounting the receptacle housing are positioned properly e Center to center of respective mounting positions is 3 35 85mm tall and 3 0 77mm wide Verify the condition of the three expose wire leads strip back to new wire if needed Connect the wire leads using the screw terminals paying strict attention to obtain the correct wiring position as it is labeled on the receptacle Green ground White common Black in the US Blue in the EU hot Single Use Bioreactor S U B 201 Appendix A Electrical Receptacle Installation Instructions Figure 2 Female Receptacle 6 If using the adapter mounting plate secure it to the selected facility electrical housing as per drawing Figure 3 otherwise proceed to step 7 3 4X 10X1 screws to mount plug to adapter p
32. Outer Support Container with a mixer water jacket and agitation control are aretemalmikerdike drive complete with control unit and an integrated into the S U B but additional electrical heater or water jacket pH or dissolved oxygen DO controls must 2 Single Use Bioreactor BPC complete be supplied by the user height diameter ratio pH amp DO Probe Autoclavable Type 12 mm diameter x 215 235 mm insertion length x 13 5 PG thread Applisens Broadley James Metler Toledo interfaces have been maintained Hold Up Volume Electrical Power Supply Requirement voltage phase amp Overall Width Overall Length Overall Height 120 240 VAC single 20 10 A 25 9 65 7 cm 23 58 4 cm 36 1 91 7 cm 36 91 4 cm 52 8 134 cm 52 5 133 4 cm Operating Temperature Range Ambient to 40 0 1 C 104 0 2 F Motor Speed 30 200 rpm Volume Range 25 50 L Maximum Bag Pressure 0 5 psi 0 03 bar Continuous Operating Time 21 days 50 L Water Jacket S U B Mixing time at nominal volume only SCIENTIFIC Part of Thermo Fisher Scientific Standard BPC Configurations SH30774 01 For use with tubing welder and CPC Quick Connect 2 Inoculum Addition Front Face N 1 Overlay Gas Sparge Back Face 7 Thermo Well Small Volume Sample 10 11 Feed Lines Standard BPC Configurations CX5 14 Film For use with tubing welder and CPC Quick Connect Description Tubing Set ID X OD
33. Run section 3 Loose items such as the drive shaft tools and RTD probes should be returned to their storage locations to prevent accidental damage Between runs the S U B hardware outer support container probe shelf drive shaft and mixer drive etc can be wiped down with sanitary wipe The outer support container can also be cleaned with typical stainless steel cleaner Store drive shaft in storage holder located near the handle of the outer support container The S U B hardware system can be cleaned to the extent of standard laboratory cleaning procedures Care should be taken to ensure electrical connections have been disconnected and electrical enclosures are closed tightly It is also recommended that excess water is not introduced under the heat shield or over the control panel A wipe down with normal disinfectant solutions is sufficient but without using excessive amounts of liquid The unit must be allowed to fully dry prior to being brought back into operation Single Use Bioreactor S U B 37 Routine Maintenance Preventive Maintenance Schedule Thermo Scientific Section 1 9 5 U B Operating Instructions Environmental conditions operating parameters and the ability of the user to adhere to standard operating procedures as outlined in this User s Guide can have significant impact upon the useful life of the S U B hardware system The following guidelines are based upon standard operating conditions
34. Section 5 2 Ordering Information Hardware 5 2 Ordering Information Hardwa re 50 L 100 L 250 Land 500 L S U B Systems Include 304 stainless Standard S U B steel outer support container with swivel caster platform variable speed agitation controller motor drive assembly with shaft PID temperature Products controller resistive only RTD sensor integrated resistive heating element or jacket probe shelf and standard tool set Part Number Description Part Number Description SV50197 01 100 L S U B Electric Resistive Heater US version 120 VAC SV50197 02 100 L S U B Electric Resistive Heater EU version 240 VAC SV50197 03 100 L S U B Water Jacket US version 120 VAC SV50197 04 100 L S U B Water Jacket EU version 240 VAC Part Number Description Part Number Description SV50200 02 500 L S U B Electric Resistive Heater US EU version 208 240 VAC SV50200 04 500 L S U B Water Jacket US EU version 208 240 VAC Thermo Scientific Single Use Bioreactor S U B 194 Section 5 2 Ordering Information Hardware 1000 L S U B Systems Include 304 stainless steel outer support container with swivel caster platform variable speed agitation controller motor drive assembly with shaft PID temperature controller three load cells with summing block display and wiring RTD sensor integrated resistive heating element probe shelf and standard tool set Part Number Description Part Number Description
35. THE EMERGENCY STOP IS ACTIVATED a 7 Verify that the main motor power switch is in the off position 8 Connect all electrical plugs to facility power NOTE 120 VAC 250 LS U B to be connected to a dedicated 20A circuit Refer to hardware electrical labels and schematics to ensure proper electrical voltage is connected to the S U B The main power switch can now be turned on 9 Resistive heating systems Verify the temperature controller is off the display should be flashing in the standby position 10 Water Jacket Systems Connect water inlet and outlet lines from temperature control unit quick connects to the jacket Figure 1 17 For 50 L through 500 L and 2000 L S U B systems the inlet is typically on the left side if facing the connectors For the 1000 L S U B the inlet is the lower connection and the outlet is the upper Single Use Bioreactor S U B 22 Air Line Preparation Porous Frit Sparge Thermo Scientific Section 1 7 Installation and Setup Figure 1 17 Water Jacket Connection Figure 1 18 Frit Spage line holder See Table 1 6 for recommended flow rates The operating pressures at the level of the S U B are of primary importance and these values must be adhered to Please note flow rates in the graphics include both half and full volume applications If you will be using a porous frit sparge in your S U B bag whether alone or in a dual sparge setup we recommend using the frit sparge line support
36. as outlined in the S U B Users Guide High wear items such as bearings seals O rings and sterilization valves common to conventional bioreactor systems have been purposefully considered in the design of the disposable construction of the S U B The S U B system is inherently robust and requires low levels of routine maintenance Taking time between bioreactor runs to clean the exterior of the S U B is certain to improve the appearance and overall longevity of the hardware system The drive motor is industrial grade induction motor with permanently sealed and lubricated gear box The drive shaft is constructed to wear slightly with use and should be visually inspected after each run The heating elements are designed to operate well within manufacturer s design limits and will provide years of consistent performance Visual inspection of wear components and following the guidelines listed below will be sufficient to ensure years of dependable service Replacement parts are available refer to Section 4 4 for information Replace drive shaft assembly after one year of service or refer to the following wear specifications e Refer to the Table 1 7 for new drive hex diameters and minimum drive shaft head hex diameters Diameters are measured at widest location across the points Replace worn shaft head assembly when drive hex diameter at widest location measures equal to or less across the points Figure 1 20 e Lightly coat drive cap thre
37. bioreactor preventing potential vent filter Th ermo Scientific blockage and reducing fluid loss due to evaporation It is offered in HyClone Single Use hoth single and double chill plate Bioreactor S U B formats a w jab N J 0 1 wl a lt Condenser System Overview The S U B Condenser System consists of 3 Peristaltic pump to return condensate saa the following components to bioreactor The HyCl t tect t filter block by on a cca Bataan is oe 1 Cart and brackets convenient means 4 Temperature control unit TCU also filters S U B BioProcess Containers BPCs are not intended to of organizing and transporting key referred to as a chiller circulates mae working elements of the Condenser water to cool the condenser plate operate under pressure and fouled blocked exhaust filters lead to ne S Condenser disiosables the BPC bag pressurization While vent filter heaters may prevent condensate ius l l l P l build up in many instances with larger bioreactors e g 2000 L S U B 2 Condenser chill plate disposable double chambered bag tubing ang this becomes less effective whereas condensing out the moisture first COUDE ape CONCENSE DANG A E NOUA WGN me al samoce eeni o preventing liquid from reaching the filters secured to the condenser plate to cool gases flow and are chilled and in which exhaust gases Up to two plates can be the condensate collects and is returned
38. cm 1 2 12 7 mm ID X 3 4 19 1 mm OD C Flex X 60 152 cm split to Pall Kleenpak Aseptic Connector Male Pall Kleenpak Emflon II 0 2 um Exhaust Filter 1 2 12 7 mm ID X 3 4 19 1 mm OD C Flex X 6 15 cm split to Bottom Drain 1 2 12 7 mm ID X 3 4 19 1 mm OD C Flex X 24 61 cm and Harvest 3 8 9 5 mm ID X 5 8 15 9 mm OD C Flex X 24 61 cm and 3 8 9 5 mm ID X 5 8 15 9 mm OD C Flex X 24 61 cm reduced to 1 8 3 2 mm ID X 1 4 6 4 mm OD C Flex X 12 30 cm 4 T E E 1 4 6 4 mm ID X 7 16 11 1 mm OD C Flex X 6 15 cm to Pall Kleenpak parg Check Valve to 1 4 6 4 mm ID X 7 16 11 1 mm OD C Flex X 60 152 cm Gamma 0 2 um 3 8 MPC Insert Plugged s Se gl 1 Ro as Data Sheet Ordering Information for Standard BPC Products Part Number Description Presentation as dry BPC systems Supplied flat packed Two polyethylene outer layers Description Product code Label Lot number Outer Packaging Expiry date on outer packaging and shipping container Sterilization Irradiation 25 to 38 kGy inside outer packaging ST melita Durable cardboard carton Documentation Certificate of Analysis provided with each lot for each delivery Custom Options Custom Hardware Options For stainless steel capabilities please contact your sales representative for HyClone products for more information 2009 Thermo Fi
39. diameter ratios mixer design and location and typical control system The Thermo Scientific HyClone Single Use Bioreactor S U B is the leading single use alternative to conventional stirred tank bioreactors for animal and insect cell culture The current range includes units with maximum working volumes of 50 100 250 500 1000 and 2000 L The S U B is now available with new optimized standard BioProcess Containers with improved dual sparge capability Open pipe and frit sparging systems are now included to provide a wider range of operating conditions This document covers S U B systems with a maximum working volume of 50 L and the new optimized standard S U B s with dual Sparge capability which provide a wider range of ka values better pH control and lower with agitator assembly sparger vent filter inlet and outlet ports plus ports for integration of sensor probes and line sets The S U B BPC is supplied sterile and ready to use foaming than the original disk sparger 3 Drive Shaft inserted into the S U B BPC The S U B BioProcess Container BPC is supplied sterilized through the mixing drive motor and locked by irradiation and therefore does not require any facility hook ups for into the disposable agitator assembly sterilization or cleaning A key element to the single use design is the 4 Process Controls temperature not on plastic polyethylene impeller with a bearing seal assembly linked to 1
40. from stainless steel e Plastic carry handle for easy transport right out of the autoclave e Positions probes on 15 incline for greater probe membrane longevity Will restrain probe bellows from collapsing during sterilization e Probe holder accommodates two probes Elevated Dual Frit Sparge Line Filter Bracket support Fi a 6 SV50200 500 L SH30774 04 SH30720 01 SV20664 03 SV50177 01 SV20750 01 SH30845 02 S U B Hardware System standard S U B BPC standard Heavy Duty Tubing Clamp 10 pack Autoclave Iray for Autoclaving Probe Kits Bioreactor Probe Assembly non sterile for use in autoclave S U B Temperature Port used to verify RTD calibration Sterile Sampling Manifold with Luer Locks 10 pack Bioreactor control system DO probe pH probe Sterile Aseptic Connection Metho Peristaltic Pump Temperature Control Unit TCU d Necessary for regulation of gas pressure and flow rate control for DO and pH set points Autoclavable Probe 13 mm X 13 5PG thread w 195 235 mm insertion length Autoclavable Probe 13 mm X 13 5PG thread w 195 235 mm insertion length Tubing Welder Terumo or Wave Steam In Place SIP Sterilizer or Laminar Flow Hood Used for fluid transfer between line sets and containers Necessary for water jacket temperature control not provided 500 L with Electric Resistive Heater The stainless steel outer support container contains the following features i Mixer motor 6
41. length x 13 5 PG thread Hold Up Volume Electrical Power Supply Requirement 120 240 VAC single 120 240 VAC single voltage phase amp 20 10 A 10A Overall Width 26 66 cm Overall Length 35 88 9 cm Overall Height 60 152 4 cm Operating Temperature Range Ambient to 40 0 1 C 104 0 2 F 250 L Water Jacket U B Mixing time at nominal volume only Part of Thermo Fisher Scientific pee Ge S os A DA E A a E O Standard BPC Configurations SH30774 03 For use with tubing welder and CPC Quick Connect 1 4 Probe Ports 4 Il Front Face ee Back Face 7 Overlay Gas Sparge 14 Porous frit gas sparge 9 10 Feed Lines Standard BPC Configurations CX5 14 Film For use with tubing welder CPC Quick Connect or Triclamp End Treatments Line Description Tubing Set ID X OD X Length End Treatment Pall Kleenpak aseptic 1 4 Probe Ports 4 1 2 12 7 mm Tube ports connectors KPCHT Series Female Thermo Well Thermo Well Adapter for 1 4 6 4 mm Diameter SterilEnz Pouch with Small Volume Sample 1 8 3 2 mm ID X 1 4 6 4 mm OD C Flex X 18 46 cm injection site assembl 1 2 12 7 mm ID X 3 4 19 1 mm OD C Flex X 60 152 cm reduced to 3 8 9 5 mm ID X 5 8 15 9 mm OD C Flex X 12 30 cm splits to Plugged 1 4 6 4 mm ID X 7 16 11 1 mm OD C Flex X 12 30 cm reduced to 3 8 MPC Insert 1 8 3 2 mm ID X 1 4 6 4 mm OD C Flex X 12
42. om w Thermo Scientific Single Use Bioreactor S U B 11 1 5 Additional System Components Probe Integration Required and Optional Accessories Thermo Scientific Section 1 5 Additional System Components The probe assembly is an innovative disposable design to package user supplied pH and DO probes for sterilization and to aseptically connect them to the S U B BPC The probe assembly Figure 1 5 includes the following components 1 Molded bellows cover 2 Threaded probe adaptor 3 Pall Kleenpak Connector KPCHT Series high temperature 4 Cable ties 4 3 J 4 j I J Figure 1 5 Probe Assembly To assist in the operation of the Single Use Bioreactor the following additional accessories are available see Section 3 for ordering information e Heavy duty tubing clamps sold separately Required typically four or five e Autoclave tray for probe kits stainless steel Required minimum of one required The autoclave support tray provides an inclined fixture for two probes during autoclaving to minimize stress on the probes and to prevent collapse of the silicone bellows e Sampling manifold with luer lock Optional e S U B temperature sample port for RTD calibration validation Optional e Vent Filter Heater System Optional For users who require additional protection for the exhaust vent filter on the standard U B BPC Consists of filter heater programmable controller and power cord Condenser Syste
43. on the front BPC panel first This way the door will allow access when connecting the last set of stretch hooks Figure 2 72 Thermo Scientific Single Use Bioreactor S U B 65 Section 2 3 single Use Bioreactor BPC Loading 1000 L Water Jacket 12 While BPC begins to fill with air remove the protective packaging from the exhaust vent filters If the bag is filled sufficiently with air the filters can be loaded into the holder Figure 2 73A If using elevated exhaust vent filters use the corresponding extended dual vent filter bracket Figure 2 73B 13 Four hanging tab hooks are located on the top of the outer support container These hooks can be used to secure the BPC from the top hanging tabs once it is inflated If connecting all four corners requires excessive stretching of the BPC it is acceptable to only use two or more hooks preferably at opposite corners Figure 2 74 Figure 2 74 Thermo Scientific Single Use Bioreactor S U B 66 Section 2 3 single Use Bioreactor BPC Loading 1000 L Water Jacket Drive Shaft Insertion The drive shaft is constructed by assembling three or four pieces these must be assembled and inserted piecewise in areas having low ceilings Reminder Operators should be elevated i e ladder to effectively assemble and insert the drive shaft 14 Verify the drive shaft sections and tools are available 15 Prepare the hollow pass through by first removing the latch pin on the safety cover
44. pH and DO control must be managed by an external controller supplied by the end user An optional exhaust vent filter heater is available for increased longevity of the exhaust filter on the S U B BPC Heating the filter sufficiently to eliminate the formation of condensation is an effective means of reducing the risk of fouling of the filter membrane The heater is factory preset to operate between 40 50 C but can easily be adjusted to the demand of the application Temperature settings above 60 C are not recommended The Condenser System is recommended for 2000 L S U B use and is also available as optional hardware for smaller systems It cools exhaust gases and re circulates the condensate into the bioreactor The S U B can be operated within the temperature range from ambient to 40 C For resistive heater systems temperature setpoints can be adjusted via the temperature controller located on the front panel of the S U B control box This controller is preprogrammed to avoid overshoot during heat up and maintain a target temperature of 0 5 C based on the set value display The process temperature is measured by means of a supplied RTD pt 100 that is inserted into the thermo well of the S U B BPC Water Jacket system temperature control is maintained through the temperature control unit TCU supplied by the end user Single Use Bioreactor S U B 5 Heating Performance External Control Thermo Scientific Load Cells
45. poly bags and bubble wrap from sparge filters Use the rear door to gain inside access to the floor of the hardware Place a clamp on the bottom drain line at this time Figure 2 100 Figure 2 97 Figure 2 98 13 Guide the sparge inlet line and filter through the bottom cutout in the outer support container e The plastic center tank insert of the floor provides the port locations for both the bottom drain Figure 2 101 and the open pipe sparge Figure 2 102 e Opening the pivoting access cover in the bottom of the tank will allow access for loading the frit sparge gas line Figure 2 103 Thermo Scientific Single Use Bioreactor S U B 75 Section 2 4 single Use Bioreactor BPC Loading 2000 L Water Jacket Figure 2 101 Figure 2 103 Figure 2 104 Three cut out holes are provided in the tank for frit sparges these holes are furthest from tank center line and align with the inside edge of this access cover door Figure 2 104 NOTE Air pressure to the sparger on the S U B BPC should not exceed 8 psi Also while a sparge line check valve is provided for each sparge line it is not uncommon for some fluid to bypass check valves during typical use We recommend elevating the sparge line filter as is feasible to help reduce this tendency A 14 Tare the load cell display before proceeding 15 Using the positioning tab pins provided secure the BPC by attaching the tabs on the bottom of the BPC onto the position tab pin
46. position moves to ground level opens both doors on the chiller plate and loads the condenser BPC from the front keeping the BPC in a saddle bag shape The vents can hang freely Figure 2 109 The second operator routes the gas inlet lines around and behind the vent holders and inspects both lines to ensure they are connected to the S U B and are not twisted or kinked adjusting them as needed Figure 2 110 Figure 2 108 Figure 2 109 Figure 2 110 4 The second operator then holds the vent filters places them into the vent filter holders above the chill plate Figure 2 111 5 The first operator then uses the Velcro staps to secure the filters in position Figure 2 112 6 The first operator then uses the grasping tabs to position the bag using the two lower button pins on each side of the chiller plate Figure 2 113 Thermo Scientific Single Use Bioreactor S U B 83 Section 2 4 single Use Bioreactor BPC Loading 2000 L Water Jacket Figure 2 111 Figure 2 112 Figure 2 113 7 The clear side doors are then closed as the operator carefully manipulates first the gas inlet line and then the gas outlet line Figure 2 114 to clear the doors as each is closed and latched Figure 2 115 8 The second operator then locates the gas inlet lines into the clips behind the vents Figure 2 116 Figure 2 114 Figure 2 115 Figure 2 116 Thermo Scientific Single Use Bioreactor S U B 84 Ther
47. provided Manipulation of the S U B BPC as the last few liters of media is removed can minimize liquid hold up within the S U B Single Use Bioreactor S U B 9 Aseptic Connections Thermo Scientific Aeration Sampling Section 1 4 BioProcess Container Characteristics Gas to liquid mass transfer in cell culture bioreactors is controlled by the solubility of the gas in the liquid its distribution and the temperature and pressure Direct air sparging is a way of providing for the oxygen requirements of mammalian cell cultures The standard S U B bag incorporates a unique disposable dual sparging design that allows for optimal aeration of the culture process and effective CO stripping Multiple aseptic connection options exist for S U B users The standard BPCs include tubing welder sections quick connects and Pall Kleenpak connections Ihe S U B BPC is designed with various lengths and dimensions of thermoplastic tubing for the purpose of addition to and dispensing from the S U B BPC Refer to Ordering Information in Section 3 for custom end treatment options The S U B is equipped with a small volume sample port that is adjacent to the S U B BPC thermo well This small diameter silicone dip tube of 6 length 15 24 cm allows low void volume samples to be taken for cell viability and density as well as analyte analysis This dip tube is supplied with an aseptic luer lock connector SmartSite that allows for direc
48. ring underneath the sleeve Figure 2 128 This is a visual indicator that the sleeve is not in a locked position Figure 2 127 Figure 2 128 Thermo Scientific Single Use Bioreactor S U B 88 Section 2 4 single Use Bioreactor BPC Loading 2000 L Water Jacket 7 Place the female side of the quick connect over the male end Figure 2 129 The connection is fully seated when the red of the male end is no longer exposed Figure 2 130 8 Slide the sleeve towards the connection allowing the push button to lock into position this will engage the locking mechanism and also cover the red indicator ring Figure 2 131 NOTE Once fully connected no red coloring should be visible This state of no visible red indicates a proper connection A F gt 129 Figure 2 130 Figure 2 131 im t Figure 2 9 Remove the latch pin load the partially assembled drive shaft through the hollow pass through and hold it in position sufficient to allow the latch pin to be inserted in to the next set of holes 10 Assemble the two middle sections of the drive shaft by joining the segment using the quick connects as performed in the previous steps 11 Obtain the upper section of the drive shaft and assemble it to the other segment in the manner described previously Once the sections are secure remove the latch pin and return the wrenches to the tool holder Thermo Scientific Single Use Bioreactor S U B 89 Secti
49. sheet E 1 9 1 15 67 39 8 cm 12 mm diameter x 215 235 mm insertion length x 13 5 PG thread 208 240 VAC single 30 A 52 132 1 cm 54 137 2 cm 126 4 321 1 cm Mixing time at nominal volume only SC Il ENTIFIC N CO N re Standard BPC Configuration SH30774 07 9 Media Fill Auxiliary Drain For use with tubing welder and CPC Quick Connects 9 Feed Li Also available without the condenser assembly as the SH30774 08 1 5 Probe Ports 5 Sn mn Front Face 10 Inoculum Addition Feed Li ga noculum Addition Feed Line 6 Thermo Well Impeller 7 Bottom Drain Harvest T Media Fill wares 11 Base Addition 12 Overlay Gas Sparge 13 Exhaust Line 14 Condensate Return Line 8 Feed Line 9 Media Fill Auxiliary Drain 15 Condenser Bag Assembly Back Face 16 Porous Frit Gas Sparge 3 18 3 Tri Clamp Port 17 Open Pipe Gas Sparge Standard BPC Configuration CX5 14 Film For use with tubing welder and CPC Quick Connects Line Description Tubing Set ID X OD X Length End Treatment TM 1 5 Probe Ports 5 1 2 12 7 mm Tube ports na ena a Pue connectors Female 10 11 7 EE Thermo Well Thermo Well Adapter for 1 4 6 4 mm Diameter SterilEnz Pouch with Small Volume Sample _ 1 8 3 2 mm ID X 1 4 6 4 mm OD C Flex X 24 61 cm injection site assembl 3 4
50. the fully up or on position Single Use Bioreactor S U B 29 Section 1 8 5 U B Operation Information Scales and Weighing Monitoring liquid volume within the S U B during operation can be Systems critical in cell culture applications that involve nutrient media feeds This can also be a useful method for increasing the scalability of the S U B by starting the process run at minimum operating volume The ability to track operating volume by use of load cells or weigh scales allows the user the ability to control liquid volume and cell density as the bioreactor is increased to rated working volume during the process run Load cells come standard on 1000 L and 2000 L S U B s An option available for S U B volumes of 50 L to 500 L is a load cell kit for weight volume measurement Load cell kits can be installed at the factory or can be added later by a certified service technician The load cell kit comes with three load cells summing block wiring and display with a choice of several interfaces For more information refer to Section 3 for ordering information Please ensure that load cells are locked down for any movement of the S U B unit Refer to Section 1 6 for proper load cell operation and care and to Appendix C for load cell calibration instructions Digital display weighing scales can be sourced from manufacturers such as Mettler Toledo Bench top scales are commonly used to measure the amount of bulk source media stored
51. to Appendix C Single Use Bioreactor S U B 21 1 7 Installation and Outer Support Container Thermo Scientific Setup Preparation Section 1 7 Installation and Setup All manual movements of mobile S U B hardware does not include 2000 L S U B should be over smooth surfaces with the S U B empty and disconnected from all power and gas feed sources All load cells must be fully locked down Refer to the appropriate subsection within Section 3 of this User s Guide for graphics illustrating the control panel interface for the appropriate hardware 1 Verify facility electrical supplies are sufficient to support the power requirements of the S U B and ancillary components such as controllers or pumps 2 Locate the outer support container in the area for the cell culture run 3 If monitoring the volume the unit may be located on a scale if load cells are not utilized Other applications may measure all liquids going in and coming out 4 Level the platform by disabling the swivel casters on the bottom of the outer support container This is accomplished by threading the leveling pads at the center of each caster to the floor 5 Verify the location of the pH DO controllers and assure cable tubing lengths are satisfactory WARNING ELECTRICAL SHOCK a 6 Verify the main power is off and the emergency stop is pulled out NOTE THE EMERGENCY STOP DISCONNECTS ALL POWER TO THE SYSTEM AN ALARM BUZZER WILL SOUND WHEN
52. to plugging the S U B into any electrical outlet Ensure the receptacle is properly earth grounded e Operating 17 C to 27 C 20 to 80 relative humidity non condensing e Storage 25 C to 65 C e Installation category II over voltage in accordance with IEC 664 e Altitude Limit 2 000 meters The Water Jacket S U B has been designed to be operated with water as the heat transfer medium with temperatures not exceeding 50 C 122 F under less than 150 psi 1 MPa operating pressure NOTE The S U B BPC operating limits for temperature are 5 to 40 C The internal pressure should not exceed 0 5 psi The water jacket is not classified as a pressure vessel and is not required to be registered inspected and stamped with the Code U symbol per section U 1 c 2 f of the ASME Boiler and Pressure Vessel Code operating with water as the heat transfer medium temperature not exceeding 99 C 210 F under less than 300 psi 2 MPa In Europe all pressure vessels and assemblies operating at pressures greater than 7 5 psi 50 MPa are subject to the Pressure Equipment Directive 97 23 EC PED As required by this directive a conformity assessment performed by the manufacture deemed this equipment as falling below the conformity assessment category 1 type of equipment vessel state of the intended fluid contents liquid fluid group of the intended contents group 2 non dangerous water As such these vessels are not deeme
53. used per system to the bioreactor Item Condenser System 115 VAC 60 Hz or 230 VAC 50 or 60 Hz Taille a DI Water or Glycol All validation testing was done with DI water a R Length 37 88 962 mm Width 23 584 2 mm Height 5 13 1908 2 mm System Weight 293 Ib 133 kg Available on pump and chiller AMID keste Upi Recommended to be activated Chiller Plate Cooling Surface Area 240 in 1548 cm Effective Bag Contact Surface Area on Chiller Plate 179 in 1155 cm Max Current Draw 115 VAC 60 Hz 11 4A Item Condenser System Operating Information rh E Min Recommended Max Exhaust Gas Flow Capacity 0 lpm lt 50 lpm 0 1 psi 95 lpm w 8 C Chiller 75 lpm w 10 C Chiller 10 C 12 rpm 30 rpm Chiller Internal Pressure Regulator IPR Set Point 7 psi lt 14 7 psi 30 psi S U B Condenser System Table 1 Condenser System Operating Parameters Part of Thermo Fisher Scientific pee Ge S os A DA E A a E O Off Gassing Management Guidelines for Single Use Bioreactor Condenser System vs Exhaust Vent Heaters Choosing the best exhaust vent configuration for Conditions 37 C batch temperature no foam in exhaust stream the S U B is application specific For most S U B configurations it will depend upon a combination of many variables e g exhaust flow rate process stability reliability expectations culture run duration or concern for evaporation loss The accompanying graph provides
54. using a counterclockwise rotation Figure 2 81 The shafts are reverse threaded to avoid loosening during operation CAUTION Do not over tighten a snug fit is sufficient Remove the latch pin gt bid Figure 2 80 Figure 2 81 Remove the latch pin load the partially assembled drive shaft through the hollow pass through and hold it in position sufficient to allow the latch pin to be inserting in to the next set of holes and allow the wrench to Single Use Bioreactor S U B 68 Thermo Scientific Section 2 3 single Use Bioreactor BPC Loading 1000 L Water Jacket be placed on the upper most flats of this shaft section Obtain the upper section of the drive shaft and assemble it to the other segment in the manner described previously Figure 2 82 Figure 2 83 Once the sections are secure remove the latch pin and return to the wrenches to the tool holder Few Figure 2 82 i Figure 2 83 19 Using two hands carefully guide the completed drive shaft into the BPC using a slight back and forth twisting motion or a counter clockwise rotation e When 2 3 5 08 cm of shaft remains twist slightly to engage the impeller e When 1 2 2 54 cm of shaft remains twist slightly to engage bearing assembly Figure 2 84 e When 0 25 0 64 cm of shaft remains twist to align motor drive keyway with one of the four outer slots on the drive shaft head 20 Ensure the head is fully seated before dire
55. 0 Thermo Fisher Scientific Inc All rights reserved FLEXMOUNT and Mettler Toledo are registered trademarks of Mettler Toledo Inc Modbus is a registered trademark of Modicon Allen Bradley is a registered trademark of Rockwell Automation DeviceNet is a registered trademark of ODVA Profibus is a registered trademark of Profibus International All other trademarks are the property of Thermo Fisher Scientific Inc and its subsidiaries Specifications terms and pricing are subject to change Not all products are available in all countries Please consult your local sales representative for details Cell Culture amp BioProcessing 925 West 1800 South In Americas Asia In Europe www thermoscientific com hyclone Logan UT 435 792 8000 32 53 85 71 80 84321 435 792 8001 fax 32 53 85 74 31 fax SCIENTIFIC Section 3 10 S U B Vent Heater Filter Data Sheet 3 10 S U B Vent Filter 3 10 1 S U B Vent Filter Heater Data Sheet Heater Data Sheet Thermo Scientific Single Use Bioreactor S U B 183 A vent filter heater system is an additional option for the Thermo Scientific HyClone Single Use Bioreactor S U B system It is supplied as an accessory for those customers who require it to protect the vent filter on the standard S U B BioProcess Container BPC D gt D N g D D gt au To ad D lt A Thermo Scientific HyClone Single Use Bioreactor Vent Filter Heater Overview The resistive heatin
56. 10 2 1 4 6 4 mm ID X 7 16 11 1 mm OD C Flex X 6 15 cm reduced to Pae ALOMON 1 8 3 2 mm ID X 1 4 6 4 mm OD C Flex X 84 213 cm rigga 3 8 9 5 mm ID X 5 8 15 9 mm OD C Flex X 84 213 cm splits to E A 12 13 E 1 4 6 4 mm ID X 7 16 11 1 mm OD C Flex X 12 30 cm reduced to a reser 1 8 3 2 mm ID X 1 4 6 4 mm OD C Flex X 12 30 cm and 3 8 MPC Body 3 8 9 5 mm ID X 5 8 15 9 mm OD C Flex X 12 30 cm 1 4 6 4 mm ID X 7 16 11 1 mm OD C Flex X 3 8 cm reduced to Hydrophobic vent filter Check Valve and 1 4 6 4 mm ID X 7 16 11 1 mm OD C Flex X 72 183 cm with Emflon Il 1 4 6 4 mm ID X 7 16 11 1 mm OD C Flex X 6 15 cm reduced to Hydrophobic vent filter 12 mm dia 25 um pores Check Valve and 1 4 6 4 mm ID X 7 16 11 1 mm OD C Flex X 72 183 cm with Emflon Il Custom BPC Options Tubing Type C Flex platinum cured silicone PVC Pharmed PharmaPure More information in selection guide of BPC catalog Tubing Size Specific lengths of 1 8 ID to 1 ID ID limitations due to port size Luers CPC Quick Connects SIP connectors Triclamp Kleenpak Lynx Refer to selection guide in BPC catalog for further SmartSite Clave Lynx steam thru CPC Steam Thru Gore steam information by connection type Note Only option valve Gore Mini TC BioQuate GE PAW SterilEnz end plug etc for probe port connections is Kleenpak Add
57. 42 6 1020 4 T T T T j 35 45 poy 37 88 962 0 Data Sheet Outer Support Container units in inches mm 2000 L S U B Water Jacket 1 2 HP EXTERNAL E STOP 15 1 RATIO GEAR MOTOR 135 00 3 479 126 42 113 MINIMUM 3211 1 CEILING HEIGHT BAG LIFT 102 42 HEIGHT 2601 5 TANK HEIGHT 50 48 47 00 1282 2 1193 8 TANK OD TANK ID 25 20 240VAC 640 1 ETL CE SMALL DOOR COMPLIANT SENSOR PROBE MOUNT 58 87 1495 3 2010 Thermo Fisher Scientific Inc All rights reserved The Thermo Scientific HyClone Single Use Bioreactor is covered under US patents USPN 7 384 783 and 7 682 067 and 7 487 688 Other US and international patents are pending SmartSite is a trademark of ALARIS Medical Systems Inc C Flex and PharMed are registered trademarks of Saint Gobain PharmaPure is a registered trademark of PharmaPure Tubing Clave is a registered trademark of ICU Medical Pall Kleenpak and Emflon are registered trademarks of Pall Corporation Lynx is a registered trademark of Millipore SterilEnz is a registered trademark of PAW Bioscience Products INC All other trademarks are the property of Thermo Fisher Scientific Inc and its subsidiaries Specifications terms and pricing are subject to change Not all products are available in all countries Please consult your local sales representative for details HyClone Laboratories Inc Logan UT USA is ISO Certified Cell C
58. 5 20 minutes before drive shaft insertion can begin times will vary based upon flow rate and inlet pressure Figure 2 46 Figure 2 46 Thermo Scientific Single Use Bioreactor S U B 54 Section 2 3 single Use Bioreactor BPC Loading 1000 L Electric Resistive Heater Four hanging tab hooks are located on the top of the outer support container These hooks can be used to secure the BPC from the top hanging tabs once it is inflated Drive Shaft Insertion The drive shaft is constructed in three or four pieces which must be assembled and inserted piecewise Reminder Operators should be elevated i e ladder to effectively assemble and insert the drive shaft 16 Prepare the hollow pass through by first removing the latch pin on the safety cover Figure 2 47 opening the safety cover Figure 2 48 and removing the threaded cap turn counterclockwise of the mixing assembly Figures 2 49 and 2 50 Figure 2 48 gt J Drive Shaft Head Hollow ee Motor Drive Keyway ad Latch Pin Figure 2 49 Safety Cover veer Drive Shaft gt Figure 2 50 Mixing Assembly Thermo Scientific Single Use Bioreactor S U B 55 Section 2 3 single Use Bioreactor BPC Loading 1000 L Electric Resistive Heater 17 Verify the three sections of the drive shaft are located in the drive shaft holders on the side of the outer support container The sections will be referred to as the upper section with the drive shaft head
59. 500 L and 1000 L U B s Tubing Type Tubing Size Pressure sensor Pendotech for 50 L to 250 L S U B comes standard on 500 L and 1000 L DO Finesse DO and pH PreSens Availability of various other disposable sensors currently pending Limited engineer to order customization possible such as additional Dependent on location in bag and compatibility media lines and vent filter lines Requires economic justification with hardware Limited customization possible as engineer to order with justification Dependent Ont OCON IN Da N E win narawate e g 1 ID port on harvest line Limited customization possible e g moving sample tempwell port to a probe tube port or swapping overlay inlet line with supplement line Dual sparger open pipe plus porous frit standard Can do an engineer to order for one or the other alone Make to order bags will be built around standard dual sparger chamber Limited customization possible Suggested use through 1 port so this is engineer to order Otherwise must use ferrule approach Typical is 10 or shorter Can use disposable capsule filter other than standard hydrophobic vent filter with Emflon Il Standard is Pall Kleenpak 0 2 micron exhaust vent filter Note Vent filter heater configuration restricts options Extended filter height above the U B bag is make to order Must be compatible with a vent filter bracket option Choice of filters for inlets used to sterile filter incomin
60. B 15 For 50 L 100 L 250 L and 500 L Volume Water Jacket Systems Thermo Scientific Section 1 6 Initial Installation Instructions a r Ta _ _ a f CENTERING R mee fo TE WASHER amp NUT a X SHOWN IN i HOLD DOWN BOLT WEIGHING POSITION BASE PLATE SECTION A A Figure 1 8 Unlocking Load Cells 6 At this point the S U B hardware is ready to be prepared for a cell culture run 7 CAUTION Do not move unit especially when filled while load cells are unlocked as this can damage the load cells 8 For load cell calibration refer to Appendix C 1 Before beginning refer to electrical schematics included with the ETP which is located on CD provided with hardware Using a flat headed screwdriver open the control box Verify position of the three way motor controller switch it should be in the middle position Figure 1 9 3 Close the control box and lock the panel using a screwdriver before continuing 4 For S U B hardware units purchased with factory installed load cells the load cells are shipped in the locked position threaded up for equipment protection Refer to Load Cell Preparation instructions above 5 At this point the S U B hardware is ready to be prepared for cell culture run Single Use Bioreactor S U B 16 1000 L Outer Support Container Preparation Thermo Scientific l Section 1 6 Initial Installation Instructions Motor Controller Switch 3 way
61. BY USING THE ASSEMBLY AID EDGE AS A GAUGE UPON COMPLETE ACTUATION A Process The Kleenpak Connector assembly is now complete and assembly aid may be removed Single Use Bioreactor S U B 106 3 1 50 L Single Use Thermo Scientific Bioreactor Section 3 specifications Section 3 Specifications This section covers the following information 3 1 I2 oro 3 4 3 5 3 6 Dl 3 8 3 9 3 10 3 11 3 12 3 1 1 I2 3 1 3 3 1 4 50 L S U B Data Sheet and Specifications 100 L S U B Data Sheet and Specifications 250 LS U B Data Sheet and Specifications 500 L S U B Data Sheet and Specifications 1000 L S U B Data Sheet and Specifications 2000 L S U B Data Sheet and Specifications Condenser System Data Sheet and Specifications Vent Filter Heater Data Sheet and Specifications Load Cells Data Sheet and Specifications Standard BioProcess Containers with Disk Sparge Systems Data Sheet Recalibration of the AC Tech Variable Speed Drive SCF SCM Models Troubleshooting 50 L S U B Data Sheet 50 L Technical Specifications Electric Resistive Heater 50 L Technical Specifications Water Jacket 50 L Control Panel Layout Single Use Bioreactor S U B 107 Thermo Scientific HyClone Single Use Bioreactor S U B 50 L The HyClone S U B provides all the advantages of single use bioprocessing without having to buy a complete new bioreactor system The critical design parameters of the S U B such as height to
62. Container is a trademark of Thermo Fisher Scientific Specifications terms and pricing are subject to change Not all products are available in all countries Please consult your local sales representative for details Cell Culture amp BioProcessing 925 West 1800 South In Americas Asia In Europe www thermoscientific com hyclone Logan UT 435 792 8000 32 53 85 71 80 84321 435 792 8001 fax 32 53 85 74 31 fax ha Gan a a a a a O Section 4 Troubleshooting Section 4 Troubleshooting Symptom Single Use Bioreactor will not operate Remedy Check power supply Verify main electrical plug connection at wall outlet verify position of main power disconnect and verify position of Emergency Stop switch Verify condition of main electrical circuit breaker of facility If protection breaker has been tripped determine fault condition The condition may exist where other electrical systems are requiring current loads beyond those allowed by the breaker S U B system should be placed on its own electrical circuit Disconnect main power cord Inspect electrical circuit breakers and fuses inside the electrical enclosure of the S U B controller Determine fault condition by visual inspection If fault condition can not be determined by visual inspection contact manufacturer Sym ptom Single Use Bioreactor temperature is below target or slow to respond Remedy Check temperature controller and sensor Thermo Scientific Verify the temperature
63. Drive shaft Zz Exhaust vent filter holder 2 Liquid sight window 3 Stainless steel 304 outer support 8 Standard tool set container g Probe access window 4 Bearing port receiver with clamp 10 Control panel D Mixing assembly with shield 11 Probe holder p Te LLA 500 L with Water Jacket The stainless steel outer support container contains the following features 1 Mixer motor 7 Liquid sight window 2 Exhaust vent filter holder 8 Standard tool set 3 Stainless steel 304 outer support g Probe access window container 10 Control panel 4 Bearing port receiver with clamp 11 Probe holder o Mixing assembly with shield 12 Probe shelf 6 Drive shaft Ld 14 15 16 ive Probe shelf Platform with swivel casters and leveling pads Resistive heater w heat shield Bottom cut outs for BPC alignment Stretch hooks ee i PTT 1 Tai Platform with swivel casters and leveling pads Laser dimple water jacket Bottom cut outs for BPC alignment stretch hooks Jacket quick connect couplings Data Sheet Outer Support Container units in inches mm 500 L S U B Electric Resistive Heater 3415 32 875 49 39 3 16 995 32 91 aF 2312 45 To al a 1955 12 EES D gt Je i 4 LP 42 15 37 1078 69 52 1 8 1323 88 500 L S U B Water Jacket 22 9 28 00 34 47 39 22 56 25 TANK HEIGHT h a 71 17 hl T i 2010 Thermo
64. Elevating the filter to ensure that it is not at the low point of the sparge line will reduce the chance that the filter is exposed to liquid a 8 Remove the bubble wrap from the bottom sparge filter Use the lower door to gain access to floor of the hardware guide the sparge inlet line and filter through the bottom cutout in the outer support container Figure 2 69 Two holes are present in the tank the hole furthest from tank centerline should be utilized with standard S U B BPC having a single sparge port a supplied cover plate is typically used to cover the second hole when using a standard BPC The operator can reach just below the tank to further extend the sparge line from the cut out Figure 2 70 T Figure 2 6 Figure 2 70 Thermo Scientific Single Use Bioreactor S U B 64 Section 2 3 single Use Bioreactor BPC Loading 1000 L Water Jacket 9 By use of the access door pass the bagged drain line set and temperature sampling port set through the large cutout in the front of the outer support container Figure 2 71 Extend the drain line set through the cutout Figure 2 71 10 Tare the load cell display before proceeding 11 Using the four stretch hooks provided secure the BPC by attaching the hooks to the hanging tabs on the bottom of the BPC Figure 2 72 Verify that the sparge filter and sparge remain in position while attaching the stretch hooks It is recommended that users secure the hanging tabs
65. Figure 1 18 for S U B sizes 250 L and smaller Simply attach this line support underneath the tank directly below where the porous frit sparge will be placed This support piece holds the sparge gas tube vertically so that the frit sparge itself is oriented in the vertical position for maximum sparge effectiveness Spring loaded screws permit a simple sliding of the holder clip onto the tank bottom With the frit sparge holder attached to the tank the sparge line can be wound through the coil of the holder to keep the sparge oriented properly WARNING The S U B BPC is not rated as a pressure vessel The BPC should not be allowed to become tight during inflation or operation Gas pressure in the BPC should not exceed 0 5 psi 0 03 bar at any time Conditions of over pressure may result in BPC damage or personal injury For reference the BPC will appear to be tight at 0 1 psi 0 007 bar a Single Use Bioreactor S U B 23 1 8 U B Operation BioProcess Container Information Preparation and Loading BPC Handling Instructions BPC Operating Thermo Scientific Information Aseptic Line Connection Section 1 8 S U B Operation Information Each outer support container is designed for a specific Single Use Bioreactor BPC Confirm the correct volume BPC is being used for the corresponding volume outer support container Sections 2 3 and 2 4 outline the installation and setup of the different volume S U B BPCs pleas
66. Fisher Scientific Inc All rights reserved SmartSite is a trademark of ALARIS Medical Systems Inc C Flex and PharMed are registered trademarks of Saint Gobain PharmaPure is a registered trademark of PharmaPure Tubing Clave is a registered trademark of ICU Medical Pall Kleenpak and Emflon are registered trademarks of Pall Corporation Lynx is a registered trademark of Millipore SterilEnz is a registered trademark of PAW Bioscience Products INC All other trademarks are the property of Thermo Fisher Scientific Inc and its subsidiaries Specifications terms and pricing are subject to change Not all products are available in all countries Please consult your local sales representative for details HyClone Laboratories Inc Logan UT USA is ISO Certified Cell Culture amp BioProcessing 925 West 1800 South In Americas Asia In Europe www thermoscientific com hyclone Logan UT 435 792 8000 32 53 85 71 80 84321 435 792 8001 fax 32 53 85 74 31 fax SCIENTIFIC 3 4 2 500 L Hardware Specifications Electric Resistive Thermo Scientific Heater Section 3 4 2 500 L Hardware Specifications Electric Resistive Heater Ambient to 40 0 1 C 104 0 2 F Asyawoay 10 9e9y ETEL siajaweieg uoneyby joujuo0s dway sauiejuoy oddns ENER Single Use Bioreactor S U B 144 3 4 3 500 L Hardware Specifications Thermo Scientific Water Jacket Sectio
67. INC All other trademarks are the property of Thermo Fisher Scientific Inc and its subsidiaries Specifications terms and pricing are subject to change Not all products are available in all countries Please consult your local sales representative for details Cell Culture amp BioProcessing 925 West 1800 South In Americas Asia In Europe Logan UT 435 792 8000 32 53 85 71 80 84321 435 792 8001 fax 32 53 85 74 31 fax www thermoscientific com hyclone 800 10 1 1068 13 LT 1398 78 55 1 18 409 58 16 1 8 COVER HEIGHT 10 22 91 07 47 Jo a da TANK JACKET HEIGHT COVER HEIGHT HyClone Laboratories Inc Logan UT USA is ISO Certified SCIENTIFIC 3 3 2 250 L Hardware Specifications Electric Resistive Thermo Scientific Heater Section 3 3 2 250 L Hardware Specifications Electric Resistive Heater Ambient to 40 0 1 C 104 0 2 F Asyawoay 10 9e9y ETEL siajaweieg uoneyby joujuo0s dway sauiejuoy oddns ENER Single Use Bioreactor S U B 134 3 3 9 250 L Hardware Specifications Thermo Scientific Water Jacket Section 3 3 3 250 L Hardware Specifications Water Jacket fanbase Impeler Scing Geele temeer O OOOO O with Exteriorlnsulation Shield Ambient to 40 0 1 C 104 0 2 F Asyawoay 10373983Y ETEL siajaweleg U018
68. OD C Flex X SSV 3 4 3 Inoculum Addition 60 152 cm for TC 72 183 cm reduced to Plugged mini TC 1 8 3 2 mm ID X 1 4 6 4 mm OD C Flex X 12 30 cm 1 2 12 7 mm ID X 3 4 19 1 mm OD C Flex X 48 122 cm reduced to 3 8 9 5 mm ID X 5 8 15 9 mm OD C Flex X 12 30 cm reduced to Media Supplement 1 4 6 4 mm ID X 7 16 11 1 mm OD C Flex X 6 15 cm splits to Addition 2 1 4 6 4 mm ID X 7 16 11 1 mm OD C Flex X 3 8 cm reduced to 1 8 3 2 mm ID X 1 4 6 4 mm OD C Flex X 12 30 cm and 1 4 6 4 mm ID X 7 16 11 1 mm OD C Flex X 12 30 cm Connections for pH Pall Kleenpak DO probes 4 1 2 12 7 mm Tube ports Aseptic Connector p KPCHT Series Female Thermo Well Thermo Well Adapter for 1 4 6 4 mm Diameter 0 Small Volume Sample 1 8 3 2 mm ID X 1 4 6 4 mm OD C Flex X 12 30 cm Smartsite Valve Port 1 2 12 7 mm ID X 3 4 19 1 mm OD C Flex X 60 152 cm reduced to 3 8 9 5 mm ID X 5 8 15 9 mm OD C Flex X 12 30 cm reduced to Bottom Drain 1 4 6 4 mm ID X 7 16 11 1 mm OD C Flex X 6 15 cm splits to Harvest 1 4 6 4 mm ID X 7 16 11 1 mm OD C Flex X 3 8 cm reduced to 1 8 3 2 mm ID X 1 4 6 4 mm OD C Flex X 12 30 cm and 1 4 6 4 mm ID X 7 16 11 1 mm OD C Flex X 12 30 cm 12 Exhaust Filter 1 1 2 3 81 cm Triclamp Port the sas Vent 3 16 4 8 mm ID X 3 8 9 5 mm OD Si
69. P supplied with each hardware system for electrical schematics Discussions regarding the Thermo Scientific Single Use Bioreactor S U B 7 Section 1 3 End User Supplied Components integration of specific bioreactor control systems for advanced control functionality should be directed though technical support staff for Thermo Scientific HyClone products and the associated controller manufacturer Controller Manufacturer Sartorius Stedim Biotech Table 1 2 Some Controller Manufacturers of S U B compatible pH DO controllers Broadley James Note The S U B will work well with any of the various control system platforms such as PLC PC DCS or proprietary operating system based controllers Thermo Scientific Single Use Bioreactor S U B 8 1 4 BioProcess Container Characteristics Single Use Bioreactor BPC Features Operating Pressure Working Volume Draining and Harvest Thermo Scientific Section 1 4 BioProcess Container Characteristics The cell culture itself is contained inside the Single Use Bioreactor BPC The chamber is manufactured from CX5 14 film which is a co extruded structure specifically designed for biopharmaceutical process usage All materials are qualified for a broad range of physical mechanical biological and chemical compatibility requirements Refer to CX5 14 data in our BPC Catalog contact your sales representative for a copy The bioreactor BPC is pre sterilized using validated
70. PreSens Availability of various other disposable sensors currently pending Limited engineer to order customization possible such as additional Dependent on location in bag and compatibility media lines and vent filter lines Requires economic justification with hardware Limited customization possible as engineer to order with justification Dependent Ont OCON IN Da N E win narawate e g 1 ID port on harvest line Limited customization possible e g moving sample tempwell port to a probe tube port or swapping overlay inlet line with supplement line Dual sparger open pipe plus porous frit standard Can do an engineer to order for one or the other alone Make to order bags will be built around standard dual sparger chamber Limited customization possible Suggested use through 1 port so this is engineer to order Otherwise must use ferrule approach Typical is 10 or shorter Can use disposable capsule filter other than standard hydrophobic vent filter with Emflon Il Standard is Pall Kleenpak 0 2 micron exhaust vent filter Note Vent filter heater configuration restricts options Extended filter height above the U B bag is make to order Must be compatible with a vent filter bracket option Choice of filters for inlets used to sterile filter incoming media or supplements Please Note Not all options are available for all ports No customization of port type and location chamber dimensions or mixing assembly is possib
71. Probe shelf 17 Jacket quick connect couplings 6 Drive shaft Data Sheet Outer Support Container units in inches mm 50 L S U B Electric Resistive Heater i 260 35 711 20 10 25 J 26 00 COVER TANK HEIGHT HEIGHT 01340 97 528 50 L S U B Water Jacket i JACKET COVER amp INSULATION NOT SHOWN 70 58 T 35 21 32 JACKET 131 29 33 92 HEIGHT 51 11 16 E bih JACKET en cove if HEIGHT 22 19 32 i 38 45 15 1 8 22 70 B 15 16 T g 7 05 aar 91 75 36 1 8 2010 Thermo Fisher Scientific Inc All rights reserved SmartSite is a trademark of ALARIS Medical Systems Inc C Flex and PharMed are registered HyClone Laboratories Inc Logan UT USA is ISO Certified trademarks of Saint Gobain PharmaPure is a registered trademark of PharmaPure Tubing Clave is a registered trademark of ICU Medical Pall Kleenpak and Emflon are registered trademarks of Pall Corporation Lynx is a registered trademark of Millipore SterilEnz is a registered trademark of PAW Bioscience Products INC All other trademarks are the property of Thermo Fisher Scientific Inc and its subsidiaries Specifications terms and pricing are subject to change Not all products are available in all countries Please consult your local sales representative for details Cell Culture amp BioProcessing 925 West 1800 South In Americas Asia In Europe www thermoscientific com hyclone Logan
72. S U B BPC as the last few liters of media drain will minimize liquid hold up within the S U B The 2000 L S U B is provided with a 1 style bottom drain near the center line of the tank bottom Temperature setpoints can be adjusted via the temperature controller located on the front panel of the S U B control box This controller is preprogrammed to avoid overshoot during heat up and maintain a target temperature of 0 5 C based on the set value SV display green LED The set value of the PID temperature controller is factory preset at 2 C to prevent premature heat up The process temperature is measured by means of a supplied RTD pt 100 that is inserted into the thermo well of the S U B BPC The temperature measured by the RTD is displayed with a red LED digital display as the present value PV For more detailed operating information on the controller refer to the ETP e Enable disable temperature controller by pushing left button e Adjust temperature set value by pushing up and down arrows located on the right e Process temperature value is illuminated in red e Flashing numeric display indicates the controller output is off standby mode Single Use Bioreactor S U B 27 Heating Performance Thermo Scientific Section 1 8 5 U B Operation Information NOTE The heaters are not enabled unless the motor agitation is operational and should not be engaged until the BPC is filled to at least the minimum
73. SV50218 09 SV50218 19 R 2000 L SV50227 09 SV50227 19 50 amp 100 L SV50194 30 SV50194 20 200 amp 250 L SV50193 30 SV50193 20 l F 500 amp 1000 L SV50218 30 SV50218 20 Panel Mount display Device Net Interface 2000 L SV50227 30 V50227 20 50 amp 100 L V50194 31 V50194 21 200 amp 250 L V50193 31 V50193 21 l l a 500 amp 1000 L SV50718 31 SV50718 71 Wall Mount display Device Net Interface 2000 L SV50227 31 SV50227 21 r D 3 aes N T Panel Mount display Modbus Plus Interface Data Sheet The 1000 and 2000 L U B s come standard with load cells The tables below list the S U B part numbers and their corresponding load cell communications platforms 1000 L S U B Systems S U B Specification Volume Description of Panel Numbers Communication Platform Electric Resistive Heater Water Jacket Heater o V50174 13 1000 L Analog Output Interface 2 SV50174 14 1000 L Profibus Interface S V50174 15 1000 L Device Net Interface J 2000 L S U B Systems S U B Specification alters Description of Panel Numbers Communication Platform Water Jacket Heater V50223 11 2000 L Modbus Plus Load Cell Interface 9V50223 12 2000 L Allen Bradley Load Cell Interface 9V50223 13 2000 L Analog Output Load Cell Interface SV50223 14 2000 L Profibus Load Cell Interface SV50223 15 2000 L DeviceNet Load Cell Interface co N x HyClone Laboratories Inc Logan UT USA is ISO Certified 201
74. Sampling Using standard luer lock on 60 mL syringe or sterile manifold l px 3 4 Ai ee ee 11 12 Thermo Scientific pnn Figure 2 154 Figure 2 155 Remove dust cover from SmartSite Figure 2 154 Clean SmartSite with sanitary wipe Connect sanitary luer lock type syringe Figure 2 155 Apply small amount of vacuum pressure by pulling out syringe plunger slightly Open pinch clamp and pull sample approximately 30 60 mL using care not to allow any back flow Close pinch clamp and remove syringe this will be a purge sample Clean SmartSite with sanitary wipe Connect sanitary luer lock type syringe Apply small amount of vacuum pressure by use of syringe Open pinch clamp and pull desired sample volume approximately 10 20 mL using care not to allow any back flow Close pinch clamp and remove syringe this will be a representative sample Clean SmartSite with sanitary wipe and replace dust cap a E a p Single Use Bioreactor S U B 98 Section 2 6 single Use Bioreactor BPC Sampling Sampling with Sterile Manifold Figures 2 156 and 2 157 Remove manifold from protective poly bag package Close all clamps on manifold lines Use sterile tubing welder to connect manifold to sample line Inspect welds and open flow path by pinching the welds Open two clamps at inlet and the clamp at purge bag 100 mL pillow bag 6 Purge sample line by filling this bag recommended 30 60 mL 7
75. Small Volume Sample Back Face 12 13 Feed Lines VG X 15 Porous frit gas sparge q9 qP 8 Exhaust Line 14 Open pipe gas sparge 4 10 Inoculum Addition Standard BPC Configurations For use with tubing welder and CPC Quick Connects Line Description Tubing Set ID X OD X Length End Treatment 1 5 Probe Ports 5 1 2 12 7 mm Tube ports connectors KPCHT Series Female ara Thermo Well Adapter for 1 4 6 4 mm Diameter SterilEnz Pouch with Small Volume Sample 1 8 3 2 mm ID X 1 4 6 4 mm OD C Flex X 24 60 cm injection site assembl 1 2 12 7 mm ID X 3 4 19 1 mm OD C Flex X 60 152 cm reduced to 3 8 9 5 mm ID X 5 8 15 9 mm OD C Flex X 12 30 cm splits to Plugged 1 4 6 4 mm ID X 7 16 11 1 mm OD C Flex X 12 30 cm reduced to 3 8 MPC Insert 1 8 3 2 mm ID X 1 4 6 4 mm OD C Flex X 12 30 cm and 3 8 9 5 mm ID X 5 8 15 9 mm OD C Flex X 12 30 cm 1 2 12 7 mm ID X 3 4 19 1 mm OD C Flex X 4 10 cm splits to Exhaust Line 1 2 12 7 mm ID X 3 4 19 1 mm OD C Flex X 10 25 cm and 1 2 12 7 mm ID X 3 4 19 1 mm OD C Flex X 10 25 cm Bottom Drain Harvest 2 Pall Kleenpak 0 2 Micron Exhaust Vent Filter 1 4 6 4 mm ID X 7 16 11 1 mm OD C Flex X 84 213 cm reduced to E Overlay Gas Sparge 1 4 6 4 mm ID X 7 16 11 1 mm OD C Flex X 6 15 cm D Le 1 8 3 2 mm ID X 1 4 6 4 mm OD C Flex X 12 30 cm meee
76. UT 435 792 8000 32 53 85 71 80 84321 435 792 8001 fax 32 53 85 74 31 fax SCIENTIFIC 3 1 2 50 L Hardware Specifications Electric Resistive Thermo Scientific Heater Section 3 1 2 50 L Hardware Specifications Electric Resistive Heater Ambient to 40 0 1 C 104 0 2 F Asyawoay 10 9e9y ETEL siajaweieg uoneyby joujuo0s dway sauiejuoy oddns ENER Single Use Bioreactor S U B 114 3 1 3 50 L Hardware Specifications Thermo Scientific Water Jacket Section 3 1 3 50 L Hardware Specifications Water Jacket with Exterior Insulation Shield 3 4 MNPT nipple with Hansen quick connect check valves Asyawoay 10373983Y ETEL siajaweleg U018 Y jonuog dwaj sauiejuoy Woddns Electrical Power Supply Requirement voltage phase amp 120 240 VAC single 10 A Operating Temperature with heater Ambient to 40 0 1 C 104 0 2 F Validated System Reliability minimum 0 9 90 HEIER Single Use Bioreactor S U B 115 Section 3 1 4 50 L Control Panel Layout 3 1 4 50 L Control Panel Layout 50 L Control Panel Electric Resistive Heater speed Control Key Pad Main Power Source 16 Pin Communication Quick Connect Motor Controller On Off Switch Temperature Controller External Heater Relay Input Main Switch a Buzzey am T Emergency Stop He
77. Users Guide i Thermo Scientific HyClone Single Use Bioreactor Part of Thermo Fisher Scientific SCIENTIFIC Thermo Scientific HyClone Single Use Bioreactor S U B User s Guide UG003 Rev 11 Preface Patents and Trademarks The Thermo Scientific HyClone Single Use Bioreactor is covered under US patents USPN 7 384 783 and 7 682 067 and 7 487 688 Other US and international patents are pending Kleenpak and Pall are registered trademarks of Pall Corporation SmartSite is a registered trademark of ALARIS Medical Systems Inc C Flex and PharMed are registered trademarks of Saint Gobain Envirotainer is a registered trademark of Envirotainer AB Broadley James and BioNet are trademarks of Broadley James Corporation Applikon is a trademark of Applikon Biotechnology DeltaV is a registered trademark of Emerson Process Management TruViu is a registered trademark of Finesse Solutions LLC CelliGen and BioFlo are registered trademarks of New Brunswick Scientific BIOSTAT is a registered trademark of Sartorius BBI Systems Terumo is a registered trademark of Terumo Meical Corporation PharmPure is a registered trademark of PharmaPure Tubing All other trademarks registered trademarks or logos are properties of their respective owners NOTE The drawings used throughout this document are representative only and may change A We have taken precautions to ensure this manual is accurate and compl
78. V50227 11 50 amp 100 L SV50194 02 SV50194 12 200 amp 250 L SV50193 02 SV50193 12 P 500 amp 1000 L SV50718 07 SV50718 17 Panel Mount display Allen Bradley Rio Interface 2000 L SV50227 02 SV50227 12 50 amp 100 L SV50194 03 SV50194 13 200 amp 250 L SV50193 03 SV50193 13 500 amp 1000 L SV50718 03 SV50718 13 Panel Mount display Analog Output Interface 2000 L SV50227 03 SV50227 13 50 amp 100 L SV50194 04 SV50194 14 200 amp 250 L SV50193 04 SV50193 14 l e 500 amp 1000 L SV50218 04 SV50718 14 Panel Mount display Profibus Interface 2000 L V50227 04 V50227 14 50 amp 100 L V50194 05 V50194 15 200 amp 250 L V50193 05 V50193 15 5 500 amp 1000 L SV50218 05 SV50218 15 Wall Mount display Modbus Plus Interface 2000 L SV50227 05 SV50227 15 50 amp 100 L SV50194 06 SV50194 16 200 amp 250 L SV50193 06 SV50193 16 l an 500 amp 1000 L SV50218 06 SV50218 16 Wall Mount display Allen Bradley Rio Interface 2000 L SV50227 06 SV50227 16 50 amp 100 L V50194 07 V50194 17 200 amp 250 L V50193 07 V50193 17 500 amp 1000 L SV50718 07 SV50218 17 Wall Mount display Analog Output Interface 2000 L SV50227 07 SV50227 17 50 amp 100 L SV50194 08 SV50194 18 200 amp 250 L SV50193 08 SV50193 18 MEDE 500 amp 1000 L SV50218 08 SV50218 18 Wall Mount display Profibus Interface 2000 L SV50227 08 V50227 18 50 amp 100 L SV50194 09 SV50194 19 200 amp 250 L SV50193 09 SV50193 19 woarde 500 amp 1000 L
79. X Length End Treatment i r Hydrophobic vent filter 1 Overlay Gas Sparge 1 4 6 4 mm ID X 7 16 11 1 mm OD C Flex X 6 15 cm with Emflon I 3 1 4 6 4 mm ID X 7 16 11 1 mm OD C Flex X 60 152 cm reduced to O Plugged mouton 1 8 3 2 mm ID X 1 4 6 4 mm OD C Flex X 12 30 cm gat Palle Kleenpak aseptic 3 6 Probe Ports 4 1 2 12 7 mm Tube ports connectors KPCHT Series Female Thermo Well Thermo Well Adapter for 1 4 6 4 mm Diameter SterilEnz Pouch with small Volume Sample 1 8 3 2 mm ID X 1 4 6 4 mm OD C Flex X 18 46 cm injection site assembl 1 2 12 7 mm ID X 3 4 19 1 mm OD C Flex X 60 152 cm reduced to 3 8 9 5 mm ID X 5 8 15 9 mm OD C Flex X 12 30 cm splits to ao 1 4 6 4 mm ID X 7 16 11 1 mm OD C Flex X 12 30 cm reduced to fee n 1 8 3 2 mm ID X 1 4 6 4 mm OD C Flex X 12 30 cm and 3 8 9 5 mm ID X 5 8 15 9 mm OD C Flex X 12 30 cm mi 1 4 6 4 mm ID X 7 16 11 1 mm OD C Flex X 6 15 cm reduced to 1 8 3 2 mm ID X 1 4 6 4 mm OD C Flex X 60 152 cm 3 8 9 5 mm ID X 5 8 15 9 mm OD C Flex X 60 152 cm splits to l aan alin 1 4 6 4 mm ID X 7 16 11 1 mm OD C Flex X 12 30 cm reduced to eh ea 1 8 3 2 mm ID X 1 4 6 4 mm OD C Flex X 12 30 cm and 3 8 MPC Body 3 8 9 5 mm ID X 5 8 15 9 mm OD C Flex X 12 30 cm Pall Kleenpak aseptic 1 2 12 7 mm ID X
80. a sheets 026 028 029 030 and 034 It is assembly linked to an external mixer drive jacket and agitation control are integrated into the recommended that all customers transition to use the The S U B consists of the following components S U B but additional pH or dissolved oxygen DO new dual sparge BPCs 1 Outer Support Container with a mixer drive controls must be supplied by the user or Bia Es Ce wy 1000 L Water i 50 L Water Jacket S U B Jacket S U B Part of Thermo Fisher Scientific ko S CUR S A DA E A a E O For use with tubing welder CPC Quick Connect or Triclamp End Treatments 4 5 Addition I p 4 Media Inoculum p Front Face iiy Vo Yy pra 4 5 Addition Media Inoculum m r m cht Impeller jt Si 11 Bottom Drain Harvest at a _ _ wiy T y pT i a 3 qh a ml yi a _ N zal 10 Thermo Well Small Volume Sample 13 Direct Gas Sparge 12 Exhaust Filter For use with tubing welder and CPC Quick Connects 1 Overlay Gas amp Sparge 3 16 4 8 mm ID X 3 8 9 5 mm OD Silicone X 4 10 cm Acro 50 Vent Filter 1 4 6 4 mm ID X 7 16 11 1 mm OD C Flex X 48 122 cm splits to 2 lines with 2 Addition 1 4 6 4 mm ID X 7 16 11 1 mm OD C Flex X 36 91 cm reduced to SmartSite Valve Port 1 8 3 2 mm ID X 1 4 6 4 mm OD C Flex X 12 30 cm 1 4 6 4 mm ID X 7 16 11 1 mm
81. a user supplied calibrated temperature bath To verify the calibration of the pressure monitor use a calibrated pressure standard Pressures can be verified by clamping the BPC inlet line and supplying gas through the overlay gas filter Expect accuracy of 0 1 psi Ihe monitor can be calibrated manually by referencing the monitor operators manual supplied in the ETP Load cells arrive uncalibrated it is recommended the load cell manufacturer or a qualified technician calibrate these systems on site Refer to Appendix C for load cell calibration instructions Expect accuracy of 0 5 kg Basic load cell default parameters are listed in the electrical schematic included with the ETP Single Use Bioreactor S U B 39 Section 2 2 single Use Bioreactor BPC Loading 50 500 L Volumes 2 2 Sing le Use Each outer support container is designed for a specific Single Use Bioreactor BPC Confirm the correct volume BPC is being used for the Bioreactor B PC corresponding volume outer support container The following section Loading for outlines the installation and setup of the S U B BPC 90 500 L Volumes 1 Remove irradiated bioreactor BPC from protective double polybags Figure 2 1 2 Load S U B BPC from the top into the outer support container Figure 2 2 3 Orient S U B BPC with bearing port up and toward motor drive with Kleenpak Connector probe ports facing bottom access cut out Figure 2 3 4 Place the bearing port into
82. ads with food grade anti seize if cap becomes difficult to turn e Replacement of the drive motor and heater element is recommended after two years of service S U B System New Hex Diameter Minimum Hex Diameter 50 100 and 250 L 0 587 0 005 14 9 mm 0 566 14 4 mm 500 and 1000 L 0 839 0 005 21 3 mm 0 820 20 8 mm 2000 L Replace after 1 year of service mandatory Table 1 7 Shaft head dimensions aa Hex diameter Z p A measurement location gt 3 Figure 1 20 Shaft head assembly Single Use Bioreactor S U B 38 2 1 Calibration Procedures Mixer Speed Calibration Temperature Controller Calibration Pressure monitor Calibration when present Load Cell Calibration when present Thermo Scientific Section 2 Calibration Procedures Section 2 Operating Information This section covers the following Dil Calibration Procedures 2D BPC Loading Instructions 50 L 100 L 250 L and 500 L 2 3 BPC Loading Instructions 1000 L 2 4 BPC Loading Instructions 2000 L 22 gt Probe Assembly 2 6 Sampling 2 7 Pall Kleenpak Connector Instructions To verify the calibration of the mixer speed use a calibrated tachometer Expect accuracy of 1 rpm Speed scaling can be modified if the calibration needs to be adjusted Please refer to Section 3 6 for details To verify the calibration of the temperature controller RTD use a S U B silicone thermo well the existing 1 8 OD RTD and
83. ailability of various other disposable sensors currently pending Limited engineer to order customization possible such as additional Dependent on location in bag and compatibility media lines and vent filter lines Requires economic justification with hardware ee Sis E ee Dependent on location in bag and fit with hardware Limited customization possible as engineer to order with justification e g 1 ID port on harvest line Limited customization possible e g moving sample tempwell port to Dependent on location in bag and fit with hardware a probe tube port or swapping overlay inlet line with supplement line Dual sparger open pipe plus porous frit standard Can do an engineer to order for one or the other alone Make to order bags will be built around standard dual sparger chamber Limited customization possible Suggested use through 1 port so Length cannot interfere with impeller and shaft this is engineer to order Otherwise must use ferrule approach Typical is 10 or shorter Can use disposable capsule filter other than standard hydrophobic vent filter with Emflon Il Standard is Pall Kleenpak 0 2 micron exhaust vent filter Note Vent filter heater configuration restricts options Extended filter height above the S U B bag is make to order Must be compatible with a vent filter bracket option Choice of filters for inlets used to sterile filter incoming media or supplements Please Note Not all options are av
84. ailable for all ports No customization of port type and location chamber dimensions or mixing assembly is possible For additional information please see the Selection Guides in the U B BPC Catalog Presentation as dry BPC systems supplied flat packed Two polyethylene outer layers Description Product code Lot number Expiry date on outer packaging and shipping container Sterilization Irradiation 25 to 38 kGy inside outer packaging Shipping Container Durable cardboard carton Documentation Certificate of Analysis provided with each lot for each delivery Electric Resistive Heater SV50171 01 50 L S U B US version 120 VAC Single Phase Includes 304 stainless steel outer support container with swivel caster platform variable speed agitation controller motor drive assembly with shaft PID temperature controller RTD sensor integrated resistive SV50171 02 50 L S U B EU version 240 VAC Single Phase heating element probe shelf and standard tool set Water Jacket SV50171 03 50 L S U B US version 120 VAC Single Phase Includes 304 stainless steel outer support container with swivel caster platform variable speed agitation controller motor drive assembly with S 50171 04 50 L S U B EU version 240 VAC Single Phase J RTD sensor integrated water jacket probe shelf and standard tool Custom Hardware Options For stainless steel capabilities please contact your sales representa
85. ains less than 20 oxygen per unit volume Table 1 6 below contains a listing of gas outlays for all sizes of S U B s to be used in specifying maximum gas flow rates for mass flow controllers or rotameters In optimal conditions no condensation or fouling the Single Use Bioreactor S U B 31 Section 1 9 S U B Operating Instructions exhaust filters have a flow capacity of at least 20 lpm at 0 1 psi The total flow rate of gas into system must be less than 20 lpm per active exhaust filter The values listed take into account the number of exhaust filters that are standard on each size of S U B 1 installed for the 50 100 and 250 L and 2 installed for the 500 and 1000 L The components of 2000 L system are scaled up to allow flow ratings of 50 lpm at 0 1 psi These values are a good rule of thumb but are not absolute requirements They are also not intended to be process gas flow settings The process gas flow settings should be adjusted as discussed below with starting conditions not exceeding 50 of the listed max values this will help avoid accidental foam out of the S U B exhaust filter S U B Range of Operating Parameters 50 L 100 L 250 L 500 L 1000 L 2000 L 2 0 40 0 0 1 Recommended Max i i i i i Gas Flow Rates 2 a E O O O O O Ar f tf 5 5 2 fof 5s 25 tof of 6 ts tof 8 fs 2 ji 15 omm YD De ee ee ee eee comm f fof pof post J i t fif fetid Nim esj o pst P
86. and agitation control are a E re drive complete with control unit and an integrated into the U B but additional electrical heater or water jacket pH or dissolved oxygen DQ controls must 2 Single Use Bioreactor BPC complete be supplied by the user Rated Liquid Working Volume 500 L Minimum Liquid Working Volume 250 L Total Reactor Volume Liquid amp Gas 660 L BPC Chamber Diameter 29 75 75 56 cm BPC Chamber Shoulder Height 57 144 78 cm Liquid Height Rated Working Volume 44 63 113 36 cm Overall Reactor Geometry height diameter ratio height diameter ratio Impeller quantity x blade count Impeller Diameter 9 88 25 09 cm pH amp DO Probe Autoclavable Type 12 mm diameter x 215 235 mm Applisens Broadley James Metler Toledo insertion length x 13 5 PG thread Hold Up Volume Electrical Power Supply Requirement 208 240 VAC single 208 240 VAC single voltage phase amp 20 A 10A Overall Width 39 22 99 61 cm Overall Length 52 12 132 38 cm Overall Height 91 231 14 cm _S U B Recommended Operating Parameters Operating Temperature Range Motor Speed Volume Range Mixing time at nominal volume only 500 L Water Jacket S0 Part of Thermo Fisher Scientific pee Ge S os A DA E A a E O Standard BPC Configurations SH30774 04 For use with tubing welder CPC Quick Connect 1 5 Probe Ports 5 Impeller Front Face 7 Bottom Drain Harvest 6 Thermo Well
87. are provided condensate must be managed by use of the condenser system or vent filter heater The S U B is equipped with a small volume sample port that is part of the S U B BPC thermo well This small diameter 1 16 x 3 16 1 59 x 4 76 mm silicone dip tube of 6 15 24 cm length allows low void volume samples to be taken for cell viability and density as well as analyte analysis This dip tube is supplied with an aseptic luer lock connector SmartSite that allows for direct sampling or attachment of various sampling manifolds by use of standard luer lock connection Alternatively manifolds can be welded onto the C Flex sample line 1 8 x 1 4 3 18 x 6 35 mm by tubing welder For recommended systems for fluid transfer see data sheet 016 contact your sales representative for a copy WARNING Gas pressure within the S U B BPC headspace should not exceed 0 5 psi 0 03 bar at any time Conditions of over pressure may result in BPC damage or personal injury A The S U B BPC is not operating as a closed system as it has both inlet and exhaust filters that are utilized to maintain a sterile environment for cells to grow without concern for contamination However conditions can be encountered when gas inlet flow rate may exceed exhaust flow rate This may be encountered in the unlikely event of the failure of a pressure regulator on a gas feed or when excessive foam within the bioreactor creates conditions of vent blockage
88. assembl 1 2 12 7 mm ID X 3 4 19 1 mm OD C Flex X 60 152 cm reduced to 3 8 9 5 mm ID X 5 8 15 9 mm OD C Flex X 12 30 cm splits to 1 4 6 4 mm ID X 7 16 11 1 mm OD C Flex X 12 30 cm reduced to 1 8 3 2 mm ID X 1 4 6 4 mm OD C Flex X 12 30 cm and 3 8 9 5 mm ID X 5 8 15 9 mm OD C Flex X 12 30 cm ae 1 4 6 4 mm ID X 7 16 11 1 mm OD C Flex X 6 15 cm reduced to Base ddion 1 8 3 2 mm ID X 1 4 6 4 mm OD C Flex X 60 152 cm lugged 3 8 9 5 mm ID X 5 8 15 9 mm OD C Flex X 60 152 cm splits to jaa S 1 4 6 4 mm ID X 7 16 11 1 mm OD C Flex X 12 30 cm reduced to oe Bottom Drain Harvest Plugged 3 8 MPC Insert 1 8 3 2 mm ID X 1 4 6 4 mm OD C Flex X 12 30 cm and aes ea 3 8 9 5 mm ID X 5 8 15 9 mm OD C Flex X 12 30 cm Y Pall Kleenpak aseptic 1 2 12 7 mm ID X 3 4 19 1 mm OD C Flex X 6 15 cm splits to connector KPCHT Series Exhaust Line 1 2 12 7 mm ID X 3 4 19 1 mm OD C Flex X 6 15 cm and Female 1 2 12 7 mm ID X 3 4 19 1 mm OD C Flex X 10 25 cm Pall Kleenpak 0 2 Micron 1 Exhaust Vent Filter 1 2 T SR 1 4 6 4 mm ID X 7 16 11 1 mm OD C Flex X 3 8 cm reduced to Hydrophobic vent filter pone parge Check Valve and 1 4 6 4 mm ID X 7 16 11 1 mm OD C Flex X 48 122 cm with Emflon I Porous frit gas sparge 1 4 6 4 mm ID X 7 16 11 1 mm OD C Flex X 6 15
89. ater Cable Motor Cable 50 L Control Panel Water Jacket Speed Control Key Pad Main Power Source Motor Controller On Off Switch Quick Connect Emergency Stop Main Switch Buzzer ae Motor Cable Thermo Scientific Single Use Bioreactor S U B 116 3 2 100 L Single Use Thermo Scientific Bioreactor al a2 32 3 3 2 4 Section 3 2 100 L Single Use Bioreactor 100 L S U B Data Sheet 100 L Technical Specifications Electric Resistive Heater 100 L Technical Specifications Water Jacket 100 L Control Panel Layout Single Use Bioreactor S U B 117 The Thermo Scientific HyClone Single Use Bioreactor S U B is the leading single use alternative to conventional stirred tank bioreactors for animal and insect cell culture The current range includes units with maximum working volumes of 50 100 250 500 1000 and 2000 L The S U B is now available with new optimized standard BioProcess Containers with improved dual Thermo Scientific sparge capability Open pipe and frit sparging systems are now HyClon e Si ngle Use included to provide a wider range of operating conditions Bioreactor S U B 100 L lala This document covers S U B systems with agitator assembly sparger vent The Therma Senie bye ones Uh naidesall ie advantaaes with a maximum working volume of 100 L filter inlet and outlet ports plus ports for and the new optimized standard S U B s with integration of sensor probes and
90. ax SCIENTIFIC 3 2 2 100 L Hardware Specifications Electric Resistive Thermo Scientific Heater Section 3 2 2 100 L Hardware Specifications Electric Resistive Heater Ambient to 40 0 1 C 104 0 2 F Asyawoay 10 9e9y ETEL siajaweieg uoneyby joujuo0s dway sauiejuoy oddns ENER Single Use Bioreactor S U B 124 3 2 3 100 L Hardware Specifications Thermo Scientific Water Jacket Section 3 2 3 100 L Hardware Specifications Water Jacket fanbase Impeler Scaing paler demeci if a with Exterior Insulation Shield check valves 5 to 40 0 1 C 104 0 2 F Single Use Bioreactor S U B 125 Asyawoay 10373983Y ETEL siajaweleg U018 Y 101 u09 dway sauiejuoy Woddns HEIER Section 1 4 100 L Control Panel Layout 3 2 4 100 L Control Panel Layout 100 L Control Panel Electric Resistive Heater Main Power Source speed Control Key Pad Motor Controller On Off Switch 16 Pin Communication Quick Connect Temperature Controller External Heater Relay Input Main Switch RTD Heater Cable Motor Cable 100 L Control Panel Water Jacket speed Control Key Pad Main Power Source ia 16 Pin Communication Quick Connect Motor Controller On Off Switch Emergency Stop Main Switch Buzzer Motor Cable Thermo Scientifi
91. bearing port receiver close door and close clamp Figure 2 4 5 Route the side and bottom ports through the side access panel and slot in tank bottom Figure 2 5 Figure 2 6 6 Direct the sparges bottom drain and sampling lines through outer support container access windows Figure 2 7 Figure 2 8 NOTE The 500 L systems have a hatch covering the access window for the bottom sparge af P T y sik gt auth Figure 2 5 Figure 2 6 Figure 2 7 Figure 2 8 Thermo Scientific Single Use Bioreactor S U B 40 Section 2 2 single Use Bioreactor BPC Loading 50 500 L Volumes 7 Connect incoming gas feed lines to both the overlay filter and the direct sparge filter Figure 2 9 Figure 2 10 a Figure 2 9 Figure 2 10 8 Inflate S U B BPC with air inflation time approximately 20 min through overlay filter but do not exceed 25 Ipm or 0 5 psi 0 34 bar internal BPC pressure As BPC inflates ensure it is properly oriented in the support container port alignment drain sparge etc WARNING DO NOT EXCEED 0 5 psi 0 034 bar within the BPC or BPC could fail For reference the BPC will appear to be tight at 0 1 psi 0 007 bar A 9 As the bag begins to fill with air manipulate the bag to align the sparge lines in the base slot Figure 2 11A Figure 2 11B NOTE While a sparge line check valve is provided for each sparge line it is not uncommon for some fluid to bypass check valves during t
92. bed previously Figure 2 54 Figure 2 55 a F Figure 2 54 Figure 2 55 19 Using two hands carefully guide the completed drive shaft into the BPC using a slight back and forth twisting motion Figure 2 56 NOTE It may be necessary for another operator to assist with drive shaft insertion As one operator inserts the drive shaft another operator should carefully manipulate the impeller when the end of the drive shaft begins to couple with the impeller a When 2 3 5 08 cm of shaft remains twist slightly to engage the impeller b When 1 2 2 54 cm of shaft remains twist slightly to engage bearing assembly c When 0 25 50 0 64 cm of shaft remains twist to align motor drive keyway with one of the four outer slots on the drive shaft head Figure 2 56 Single Use Bioreactor S U B 57 Section 2 3 single Use Bioreactor BPC Loading 1000 L Electric Resistive Heater 20 Directly couple the drive shaft to the motor by placing the threaded cap back on the hollow pass through and tighten 21 Tighten cap by placing spanner wrench counterclockwise on hollow pass through and tighten using supplied torque wrench Figure 2 57 NOTE The torque wrench is a standard 3 8 square drive and it is calibrated at the factory at 150 in lbs Figure 2 57 22 Verify wrenches have been removed from the system and returned to the storage holders 23 Close safety access cover and insert latch pin Securing
93. c Single Use Bioreactor S U B 126 3 3 250 L Single Use Thermo Scientific Bioreactor 3 3 1 3 9 2 Soe 3 3 4 Section 3 3 250 L Single Use Bioreactor 250 L S U B Data Sheet 250 L Technical Specifications Electric Resistive Heater 250 L Technical Specifications Water Jacket 250 L Control Panel Layout Single Use Bioreactor S U B 127 The Thermo Scientific HyClone Single Use Bioreactor S U B is the leading single use alternative to conventional stirred tank bioreactors for animal and insect cell culture The current range includes units with maximum working volumes of 50 100 250 500 1000 and 2000 L The S U B is now available with new optimized standard BioProcess Containers with improved dual Thermo Scientific sparge capability Open pipe and frit sparging systems are now HyClon e Si ngle Use included to provide a wider range Bioreactor S U B 250 L of operating conditions This document covers S U B systems with with agitator assembly sparger vent The HyClone S U B provides all the advantages of single use a maximum working volume of 250 L and the filter inlet and outlet ports plus ports for bioprocessing thani having to buy a complete new bioreactor new optimized standard S U B s with dual integration of sensor probes and line sets system The critical design parameters of the S U B such as height to sparge capability which provide a wider range The S U B BPC is supplied sterile and diame
94. c Single Use Bioreactor S U B 61 Thermo Scientific Section 2 3 single Use Bioreactor BPC Loading 1000 L Electric Resistive Heater 40 Insert RTD or selected temperature sensor into the thermo well Figure 2 63 a Place a small amount of glycerol 0 5 mL in well to aid in heat transfer b Sensor should be inserted until base of RTD meets the mouth of the thermo well c Secure by twisting the luer lock collar if provided the thermo well will stretch slightly when RTD is seated Figure 2 64 Figure 2 63 Figure 2 64 NOTE Verify all port clamps are closed and located as near as possible to the body of the S U B BPC a Single Use Bioreactor S U B 62 Loading 1000L_ six Water Jacket Section 2 3 single Use Bioreactor BPC Loading 1000 L Water Jacket Open both the upper and lower doors on outer support container and reach inside to open clamp on bearing port receiver located below the motor Figure 2 65 If attached remove the probe shelf from the front of the tank cut out and store on the platform Remove irradiated S U B BPC from protective double polybags Do not remove the polybags from the line sets at this stage as the BPC may become difficult to manage Do not allow the BPC or line sets to touch or drag on the floor Load the BPC through the top access door orient the bottom of the BPC first with the bearing port facing upward Figure 2 66 Figure 2 65 Figure 2 66 4 Place the top line sets
95. cell lines in the S U B system please contact technical support l 2 3 Select desired line set from the S U B BPC for fluid introduction Make aseptic connection tubing welder quick connect or tri clamp and begin liquid fill Once approximately 20 30 L of volume has been added verify the position of the BPC in the outer support container particularly the sparger and the drain line Adjust positioning if necessary for proper fit It is recommended that the top corners of the BPC be pulled upward to reduce wrinkles during filling NOTE The BPC must be pulled at the top corners as the bag will generally form wrinkles during liquid fill If the film is not pulled from the top to remove wrinkles film tensioning below the bearing port will result The stretch hooks can be removed from bottom corners of the S U B BPC when fluid volume reaches 30 volume This will eliminate any interference with the load cell readings Fill to desired liquid volume 50 100 of rated volume is recommended If employing a liquid batch to tank grounding cable with the stainless steel connector of the sample line the sample line should be purged of air prior to probe calibration Once the media has reached half volume of the S U B the agitation control can be turned on via the motor ON OFF switch Using the arrow keys adjust the setpoint speed to the desired level the red LED display indicates the stirring speed in RPM Adjust desired a
96. cketed 1000 L Jacketed 2000 L 1000 L minimum 2000 L Table 1 5 Approximate heating times for the S U B Protective earthing for the S U B hardware system and controller is provided through the ground terminal of the plug providing power Source power to the controller must provide protective earthing to this terminal in order to minimize the hazard of a possible shock in the occurrence of a fault condition Please refer to Appendix A Electrical components of the S U B are equipped with circuit protection The variable frequency drive used to power the mixer motor is protected by the use of midget quick blow fuses Ferraz Shuwmut 10A for 50 100 and 250 L OTM10 and 15 A for 500 L 1000 and 2000 L OTM15 Other components of the resistive heating system such as the temperature controller and heating element are protected with resetable breakers In the case of an electrical fault condition these safety devices are designed to protect the user from electrical shock and prevent electrical system components from being damaged Fuses can be replaced and or the breakers reset once the fault condition is resolved NOTE e The normal on setting for these breakers is in the up position e A tripped breaker will be in the mid position e Off is in the fully down position e To reset a tripped breaker it must first be moved from the tripped or mid position to the off or fully down position before moving it
97. cm reduced to Hydrophobic vent filter 12 mm dia 25 um pores Check Valve and 1 4 6 4 mm ID X 7 16 11 1 mm OD C Flex X 48 122 cm with Emflon Il Custom BPC Options C Flex platinum cured silicone PVC Pharmed PharmaPure More information in selection guide of BPC catalog specific lengths of 1 8 ID to 1 ID D limitations due to port size Luers CPC Quick Connects SIP connectors Triclamp Kleenpak Lynx Refer to selection guide in BPC catalog for further SmartSite Clave Lynx steam thru CPC Steam Thru Gore steam information by connection type Note Only option valve Gore Mini TC BioQuate GE PAW SterilEnz end plug etc for probe port connections is Kleenpak Additional ports second row of 4 for 50 L to 250 L S U Bs second row of 5 for 500 L and 1000 L U B s Pressure sensor Pendotech for 50 L to 250 L S U B comes standard on 500 L and 1000 L DO Finesse DO and pH PreSens Availability of various other disposable sensors currently pending Limited engineer to order customization possible such as additional Dependent on location in bag and compatibility media lines and vent filter lines Requires economic justification with hardware Dependent on location in bag and fit with hardware Limited customization possible as engineer to order with justification e g 1 ID port on harvest line Limited customization possible e g moving sample tempwell port to a probe tube por
98. connect couplings Data Sheet Outer Support Container units in inches mm 100 L S U B Resistive 1219 08 RS 625 1279 4 anes HEATER aos 1a 734 1 HEIGHT 1425 73 908 54 108 28 904 COVER NOT SHOWN 1087 01 l 42 008 l 100 L S U B Water Jacket JACKET COVER HOT SHOWN 663 4 26 12 JACKET HEIGHT B49 7 1425 541 l rA a4 VERAJ 94 72 TANK W L l pea JACKET HEIGHT COVER HEIGHT ia r tall T T 2010 Thermo Fisher Scientific Inc All rights reserved SmartSite is a trademark of ALARIS Medical Systems Inc C Flex and PharMed are registered HyClone Laboratories Inc Logan UT USA is ISO Certified trademarks of Saint Gobain PharmaPure is a registered trademark of PharmaPure Tubing Clave is a registered trademark of ICU Medical Pall Kleenpak and Emflon are registered trademarks of Pall Corporation Lynx is a registered trademark of Millipore SterilEnz is a registered trademark of PAW Bioscience Products INC All other trademarks are the property of Thermo Fisher Scientific Inc and its subsidiaries Specifications terms and pricing are subject to change Not all products are available in all countries Please consult your local sales representative for details Cell Culture amp BioProcessing 925 West 1800 South In Americas Asia In Europe www thermoscientific com hyclone Logan UT 435 792 8000 32 53 85 71 80 84321 435 792 8001 fax 32 53 85 74 31 f
99. ctly coupling the drive shaft to the motor by placing the threaded cap back on the hollow pass through and tightening it NOTE The cap should be easy to install when the drive shaft head is fully engaged in the hollow pass through Otherwise repeat all steps of 19 before installing cap Figure 2 84 Single Use Bioreactor S U B 69 21 22 ZO Securing Access Doors 24 25 26 Section 2 3 single Use Bioreactor BPC Loading 1000 L Water Jacket Directly couple the drive shaft to the motor by placing the threaded cap back on the hollow pass through and tighten Tighten cap by placing spanner wrench counterclockwise on hollow pass through and tighten using supplied torque wrench Figure 2 85 NOTE The torque wrench is a standard 3 8 square drive and it is calibrated at the factory at 150 in lbs Figure 2 85 Verify wrenches have been removed from the system and returned to the storage holders Close safety access cover and insert latch pin The access doors should be secured first with the latches Proper tension is obtained by adjusting the threaded latch pin Tension of the latch is adjusted by varying the position of the pin on the threaded shank The proper latch tension can be obtained by a combination of feel and visual inspection When closing the latch the handle should begin to provide resistance to closing when the leading edge of the safety pin pass through of the latch handle aligns with
100. ctor Geometry height diameter ratio Fluid Geometry Working Volume height diameter ratio mpeller quantity x blade count pH amp DO Probe Autoclavable Type Applisens Broadley James Metler Toledo insertion length x 13 5 PG thread Hold Up Volume oane mass ont ae Overall Width Overall Length Overall Height S U B Recommended Operating Parameters 100 L Water Jacket S U B Mixing time at nominal volume only Impeller Diameter 5 75 14 6 cm Part of Thermo Fisher Scientific pee Ge S os A DA E A a E O Standard BPC Configurations SH30774 02 For use with tubing welder and CPC Quick Connect 2 Inoculum Addition Front Face N 1 Overlay Gas Sparge N 0J el re Back Face 10 11 Feed Lines Standard BPC Configurations CX5 14 Film For use with tubing welder and CPC Quick Connect Line Description Tubing Set ID X OD X Length End Treatment Hydrophobic vent filter A 1 Overlay Gas Sparge 1 4 6 4 mm ID X 7 16 11 1 mm OD C Flex X 6 15 cm with Emflon I p 1 4 6 4 mm ID X 7 16 11 1 mm OD C Flex X 60 152 cm reduced to O Pgd D ae 1 8 3 2 mm ID X 1 4 6 4 mm OD C Flex X 12 30 cm aged Pall Kleenpak aseptic 1 2 12 7 mm Tube ports connectors KPCHT Series Female Thermo Well Adapter for 1 4 6 4 mm Diameter SterilEnz Pouch with Small Volume Sample 1 8 3 2 mm ID X 1 4 6 4 mm OD C Flex X 18 46 cm injection site
101. d also be reduced as much as possible Too much gas creates excess foam and higher shear conditions Provide only the level of agitation needed low viability and lysed cells reduce agitation speed cell aggregation and settling increase agitation Media formulation can also have a large affect on cell culture growth in the S U B Surfactants such as Pluronic decrease shear and increase k a but at a cost of increased foaming Thermo Scientific can offer custom media especially for the S U B and customer s specific cell line s Sym ptom Excessive foam in the bioreactor headspace Remedy Alter the liquid surface tension related to the culture media and or sparge gas A media supplement of antifoam can be used in the S U B These serve to lower the surface tension of the media and will reduce the presence of foam High sparge rates of air can result in the presence of excessive foam Testing has shown that sparging with oxygen will typically result in dramatic reduction of foam in the headspace Single Use Bioreactor S U B 191 Thermo Scientific Section 5 Ordering Information Section 5 Ordering Information This section covers the following information 5 1 D2 5 3 5 4 5 5 5 6 D Ordering Information Disposables BPC Ordering Information Hardware Ordering Information Accessories Ordering Information Partial Spare Parts List Replacement and Spare Parts Ordering Instructions Ordering Support Contact Informa
102. d leveling pads Resistive heater w heat shield Bottom cut outs for BPC alignment Stretch hooks 250 L with Water Jacket The stainless steel outer support container contains the following features 1 Mixer motor 7 Liquid sight window 2 Exhaust vent filter holder 8 Standard tool set 3 Stainless steel 304 outer support g Probe access window container 10 Control panel 4 Bearing port receiver with clamp 11 Probe holder o Mixing assembly with shield 12 Probe shelf 6 Drive shaft 14 15 16 ive Platform with swivel casters and leveling pads Laser dimple water jacket Bottom cut outs for BPC alignment stretch hooks Jacket quick connect couplings Data Sheet Outer Support Container units in inches mm 250 L S U B Electric Resistive Heater i 745 62 29 4 864 01 H T Az l 250 L S U B Water Jacket 74 85 57 15 29 15 3 22 1 2 162 34 315 18 OVERALL 1 HEIGHT i 59 69 G23 1 2 2010 Thermo Fisher Scientific Inc All rights reserved SmartSite is a trademark of ALARIS Medical Systems Inc C Flex and PharMed are registered trademarks of Saint Gobain PharmaPure is a registered trademark of PharmaPure Tubing Clave is a registered trademark of ICU Medical Pall Kleenpak and Emflon are registered trademarks of Pall Corporation Lynx is a registered trademark of Millipore SterilEnz is a registered trademark of PAW Bioscience Products
103. d as being subject to a conformity assessment and are thereby only required to meet the Sound Engineering Practice SEP guidelines outlined in the directive Installation and operation of the S U B must adhere to the manufacturer s operating guidelines in order to ensure compliance with this assessment to ensure utmost safety it is recommended that all S U B jacket systems be operated at 75 psi or less If regulatory certification is deemed necessary by a governing body or the installation is likely to require function outside of the above mentioned parameters a regulatory stamped jacket can be obtained for an additional fee Single Use Bioreactor S U B 2 1 1 Introduction Thermo Scientific Section 1 1 S U B Overview Section 1 Single Use Bioreactor S U B Overview This section covers the following information 1 1 Introduction to the Single Use Bioreactor 12 Hardware Characteristics 13 End User Supplied Components 1 4 BioProcess Container Characteristics 1 5 Additional System Components 1 6 Initial Installation Instructions 1 7 Installation and Setup 1 8 S U B Operating Information 1 9 S U B Operating Instructions The Thermo Scientific HyClone Single Use Bioreactor S U B has been designed to be a disposable alternative to conventional stirred tank bioreactors currently utilized in animal cell culture Based on years of accepted stirred tank reactor STR design the S U B emulates STR scalability and opera
104. d assembly for the 1000 and Hardware 2000 L S U B units can be found in the Integrator Reference Guide Thermo Scientific Single Use Bioreactor S U B 13 Site Preparation Electrical Connections Outer Support Container Preparation A A For 50 L 100 L 250 L and 500 L Volume Electric Resistive Heating Thermo Scientific Systems Section 1 6 Initial Installation Instructions The S U B hardware cannot be used on circuits equipped with GFCI circuit protection because of the potential for nuisance tripping The electrical plug on the S U B is a connector that offers a secure ground These connectors meet the electrical safety codes for portable equipment and are International Electrical Code IEC rated meet IEC standard 60309 This plug serves to provide electrical ground prior to power connection The supplied electrical receptacle should be hardwired into the facility by a qualified electrical technician for US installations the receptacle will require the use of an adapter mounting plate supplied which will fit into a two gang box For additional information on the adapter mounting plate please see the ETP Alternatively the system can be hardwired directly into the facility NOTE The yellow plug and receptacle are for 120VAC and the blue are for 240VAC S U B s Each outer support container is shipped directly from the manufacturer and arrives with various safety mechanisms in place Please follow the
105. d off during an E stop Examples are recording devices sensors controllers etc The max for each receptacle is 2 amps Parameter Input Mode Increase Speed Parameters Forward Reverse Decrease Speed Parameters Button Disabled Motor On Motor Off Button Disabled Thermo Scientific Single Use Bioreactor S U B 167 Section 3 7 standard BioProcess Containers with Disk Sparge Systems Data Sheet 3 7 Standard ee Standard BioProcess Containers with Disk Sparge BioProcess Systems Data Sheet Containers with Disk Sparge Systems Data Sheet Thermo Scientific Single Use Bioreactor S U B 168 The Thermo Scientific HyClone Single Use Bioreactor S U B is the leading single use alternative to conventional stirred tank bioreactors for animal and insect cell culture The current range includes units with maximum working volumes of 50 100 250 500 1000 and 2000 L The S U B is now available with new optimized standard BioProcess Containers with improved dual Thermo Scientific HyClon e sparge capability Open pipe and frit sparging systems are now included i i to provide a wider range of operating Single Use Bioreactor S U B conditions The 2000 L S U B BPC is w jab N J 0 1 gt N a lt Standard BioProcess Containers with Disk not available with the Disk Spage Sparge System System Overview The HyClone S U B provides all the advantages complete with cont
106. e Logan UT 435 792 8000 32 53 85 71 80 84321 435 792 8001 fax 32 53 85 74 31 fax SCIENTIFIC 3 5 2 1000 L Hardware Specifications Electric Resistive Thermo Scientific Heater Section 3 5 2 1000 L Hardware Specifications Electric Resistive Heater Ambient to 40 0 1 C 104 0 2 F Asyawoay 10 9e9y ETEL siajaweieg uoneyby joujuo0s dway sauiejuoy oddns ENER Single Use Bioreactor S U B 155 3 5 3 1000 L Hardware Specifications Thermo Scientific Water Jacket Section 3 5 3 1000 L Hardware Specifications Water Jacket fenbaes A npr Scing Gnpalerdaneternkdaneey B with Exterior Insulation Shield Nominal Reatng ooing ime SaTO Ambient to 40 0 1 C 104 0 2 F Asyawoay 10373983Y ETEL siajaweleg U018 Y 101 u09 dway sauiejuo4 yoddns TIENER Single Use Bioreactor S U B 156 Section 3 5 4 1000 L Control Panel Layout 3 5 4 1000 L Control Panel Layout Speed Control Main Power Source Key Pad Motor Cable Motor Controller On Off switch 7 Pressure Heater Cable Temperature Transducer Controller a i G bs Emergency Load Cell ESAE Main Switch Stop Output Fors De Pressure alarm relay Pressure Monitor Een hs lt i Display H Load Cell Input from ot External Heater summing block Load Cell 3 Relay Input Display h
107. e Phase Includes 304 stainless steel outer support container with swivel caster _ platform variable speed agitation controller motor drive assembly with i shaft PID temperature controller RTD sensor integrated resistive heating SV50197 02 100 L S U B EU version 240 VAC Single Phase elemeit arobe choli tandari toolset Water Jacket SV50197 03 100 L S U B US version 120 VAC Single Phase Includes 304 stainless steel outer support container with swivel caster platform variable speed agitation controller motor drive assembly with SV50197 04 100 L S U B EU version 240 VAC Single Phase shaft RTD sensor integrated water jacket probe shelf and standard tool set Custom Hardware Options For stainless steel capabilities please contact your sales representative for HyClone products for more information parea Used to package the pH and DO probes not supplied for sterilization PBO AOD A vas Maly Mess I and to aseptically connect them to the S U B BPC See Figure 1 SV20664 01 Heavy Duty Tubing Clamp SV20664 02 Heawy Duty Tubing Clamp 4 pack of SV20664 01 er clamp used on probe ports when connecting probe SV20664 03 Heavy Duty Tubing Clamp 10 pack of SV20664 01 SV20750 01 5 U B Temperature Sample Port Used for RTD calibration validation SV50177 01 Autoclave Tray for Probe Kits stainless steel e Kay Dog aS anc DIOS TO ESAE NAUDE ANEO DEE SH30845 01 Sterile Sampling Manifold with luer lock individ
108. e adhere to these instructions in the order in which they are presented Take care if using a sharp object when opening outer polybags to avoid damaging the S U B bag When placing bags in containers do not drag bag over corners or sharp objects Do not lift the bag by the corners or top seams Carefully coil tubing on top of bag to prevent puncturing bag with cable ties or clamps Use cushioning between tubing and bag in storage and transport The most common recommended process for making connections to the tubing lines is with an aseptic tubing fuser such as those in Table 1 4 NOTE Other connection options are available as a custom S U B BPC assembly Following the recommended tubing welder operating instructions successful connections can be obtained for filling supplementing sampling or dispensing from the S U B BPC as needed Commonly Used Tubing Welders for Aseptic Connections Tubing Brand Model die set Tubing Size ID OD Material soea 18 1 1318 695mm Table 1 4 Some tubing welders for S U B aseptic connections Single Use Bioreactor S U B 24 Working Volume Aeration Exhaust vent filter Thermo Scientific Section 1 8 5 U B Operation Information Each S U B is designed for a working volume range The minimum working volume and the rated working volume are listed in the specification table see Section 3 The total volume listed includes the headspace needed for proper aeration and gas mana
109. e as the BPC may become difficult to manage Do not allow the BPC or line sets to touch or drag on the floor Load the BPC through the rear access door orient the bottom of the BPC into the door first with the bearing port facing upward Figure 2 92 Keep the BPC folded as supplied in the packaging it will allow the bag to unfold naturally when lifted by the hoist Figure 2 92 Using the rear door for access connect the retainer hooks of the hoist to the top of the BPC via the hanging tabs starting with the furthermost two tabs Figure 2 93 and finishing with the closer two tabs Figure 2 94 Raise the BPC using the pneumatic lift one operator observing from above with the second operator at ground level controlling the lifting valve Figure 2 95 Raise the hoist until the lift reaches full stroke Once lifting stops the top level operator should hold the hoist frame and apply some minimal lifting force This will assist the lifting device to pull in any remaining slack in the cable and ensure the lift device has been fully raised Place the valve in the stop position Single Use Bioreactor S U B 73 Section 2 4 single Use Bioreactor BPC Loading 2000 L Water Jacket Figure 2 93 Figure 2 95 7 Place the top line sets still in polybags onto the lifting frame or over the side of the outer support container This will help support the weight of the BPC and also keep the BPC from being restricted during the air infla
110. e reused multiple times It needs to be stored in clean and dry conditions between each use Connect both halves of Kleenpak connector together a Lift and pull tab of protective caps to remove caps from connectors Step 3 1 a Protective caps Step a J b Hold the barrel of the larger male connector above base e Align the smaller female connector with the larger male connector e Flat sides should be aligned e Both peel away strips must remain folded and oriented in the same direction Step b Female 7 Ro A SS Connector A 2 Anti actuation A ring Flat side A Barrel CX where both folded eZ PAD over peel a way Zs K strips e xit o gt S Male Connector NOTE If the connectors are not aligned properly the connection cannot be made A c Once aligned correctly press the two connectors together firmly until both sets of locking clips snap together tightly Step c piel Locking clips not visible Locking clips WARNING A KLEENPAK CONNECTOR WILL NOT FIT INTO THE ASSEMBLY AID IF THIS CONNECTION IS NOT PROPERLY SECURED A Single Use Bioreactor S U B 104 Section 2 7 Pall Kleenpak Connector Instructions Remove Anti Actuation Device d While holding the Kleenpak Connector securely in hand pull the anti actuation ring outward Remove the anti actuation ring from the male connector Step d Install into Assembly Aid e Place connected Kleenpak Connector unit f
111. e threaded up for equipment protection Figure 1 15 and 1 16 A 15 16 open box wrench is recommended for unlocking the load cells To unlock the load cells loosen the centering washer and nut 5 8 nut until the nut is threaded down on the hold down bolt about half way Next loosen the jam nut on the base plate until the jam nut and centering washer nut meet Tighten both nuts together Using an open end wrench loosen the hold down bolt by moving it upward to provide clearance below and above the cutout in the Motor Controller Switch 3 way Pressure Outlet 240 VAC Continuous Power Non E Stoppable rere ttt SME Outlet 240 VAC Continuous Power E Stoppable Figure 1 15 Control Box for 2000 L Jacketed S U B Thermo Scientific Single Use Bioreactor S U B 20 Thermo Scientific Section 1 6 Initial Installation Instructions top plate Figure 1 16 Section A A The clearance around above and below this nut should be from 1 8 to 3 16 Once the proper clearance has been achieved retighten the jam nut SARI NIUE WASHER amp NUT ae 7 SHOWN IN 4 HOLD DOWN BOLT WEIGHING POSITION he BASE PLATE Pi SECTION A A Figure 1 16 Unlocking Load Cells 6 At this point the S U B hardware is ready to be prepared for a cell culture run 7 CAUTION Do not move unit especially when filled while load cells are unlocked as this can damage the load cells 8 For load cell calibration refer
112. eads S 50191 13 NEMA rated Vent Heater w Programmable Controller Includes low temp alarm preset temp 50 C 200 240VAC power cord power cord w flying leads NEMA rated 48 series vent filter heater kits for use with Meissner Ultracap 10 vent filters sold as a kit which includes vent filter heater controller with a water tight enclosure quick connects and installation power cord Part Number Description Additional Information SV50191 16 NEMA rated Vent Heater w Programmable Controller Includes low temp alarm preset temp 50 C 100 120VAC 20 NEMA 5 15 power cord for U S Japan SV50191 17 NEMA rated Vent Heater w Programmable Controller Includes low temp alarm preset temp 50 C l 200 240VAC 20 BS1363 power cord for United Kingdom S V50191 18 NEMA rated Vent Heater w Programmable Controller Includes low temp alarm preset temp 50 C 200 240VAC 20 CEE7 7 power cord for Europe S V50191 19 NEMA rated Vent Heater w Programmable Controller Includes low temp alarm Preset temp 50 C 200 240VAC 12 IEC320 power cord for 2000 L SUB control box HyClone Laboratories Inc Logan UT USA is ISO Certified 2010 Thermo Fisher Scientific Inc All rights reserved The Thermo Scientific HyClone Single Use Bioreactor is covered under US patents USPN 7 384 783 and 7 682 067 and 7 487 688 Other US and international patents are pending HyClone and BPC are registered trademarks of Thermo Fisher Scientific BioProcess
113. ee F 1 4 6 4 mm ID X 7 16 11 1 mm OD C Flex X 6 15 cm reduced to Plugged ae 1 8 3 2 mm ID X 1 4 6 4 mm OD C Flex X 84 213 cm ogee 3 8 9 5 mm ID X 5 8 15 9 mm OD C Flex X 84 213 cm splits to A A 12 13 E 1 4 6 4 mm ID X 7 16 11 1 mm OD C Flex X 12 30 cm reduced to A ase 1 8 3 2 mm ID X 1 4 6 4 mm OD C Flex X 12 30 cm and 3 8 MPC Body 3 8 9 5 mm ID X 5 8 15 9 mm OD C Flex X 12 30 cm 1 4 6 4 mm ID X 7 16 11 1 mm OD C Flex X 3 8 cm reduced to Check Valve and 1 4 6 4 mm ID X 7 16 11 1 mm OD C Flex X 72 183 cm with Emflon Il 1 4 6 4 mm ID X 7 16 11 1 mm OD C Flex X 6 15 cm reduced to 12 mm dia 25 um pores Check Valve and 1 4 6 4 mm ID X 7 16 11 1 mm OD C Flex X 72 183 cm with Emflon Il Custom BPC Options C Flex platinum cured silicone PVC Pharmed PharmaPure More information in selection guide of BPC catalog specific lengths of 1 8 ID to 1 ID ID limitations due to port size Luers CPC Quick Connects SIP connectors Triclamp Kleenpak Lynx Refer to selection guide in BPC catalog for further SmartSite Clave Lynx steam thru CPC Steam Thru Gore steam information by connection type Note Only option valve Gore Mini TC BioQuate GE PAW SterilEnz end plug etc for probe port connections is Kleenpak Additional ports second row of 4 for 50 L to 250 L S U Bs second row of 5 for
114. eed Thermo Scientific Single Use Bioreactor S U B 157 Section 3 6 2000 L Single Use Bioreactor 36 2000 L Single Use 3 6 1 2000 L S U B Data Sheet 6 2000 L Technical Specifications Water Jacket Bioreactor 3 6 3 2000 L Control Panel Layout Thermo Scientific Single Use Bioreactor S U B 158 Thermo Scientific HyClone Single Use Bioreactor S U B 2000 L The HyClone S U B provides all the advantages of single use bioprocessing without having to buy a complete new bioreactor system The critical design parameters of the S U B such as height to diameter ratios mixer design and location and typical control system interfaces have been maintained The S U B BioProcess Container BPC is supplied sterilized by Irradiation and therefore does not require any facility hook ups for Sterilization or cleaning A key element to the single use design is the plastic Kynar impeller with a bearing seal assembly linked to an external mixer drive 2000 L Water Jacket S U B Part of Thermo Fisher Scientific The Thermo Scientific HyClone Single Use Bioreactor S U B is the leading single use alternative to conventional stirred tank bioreactors for animal and insect cell culture The current range includes units with maximum working volumes of 50 100 250 500 1000 and 2000 L The S U B is now available with new optimized standard BioProcess Containers with improved dual sparge capability Open pipe and frit s
115. erify the probe tip is not touching the membrane 6 Place the probe assembly with probe into the autoclave tray for probe kits Figure 2 148 7 Autoclave the probe assembly using a validated sterilization cycle approximately 30 minutes at 122 C Thirty minute sterilization cycles are generally sufficient Options of wet or dry cycle parameters can be used Slow exhaust cycles are preferred as this minimizes stress on the probes during the temperature and pressure changes of autoclaving 8 Allow sufficient time for probe assemblies to cool completely before seeking to connect to the S U B bag for probe insertion 9 When stored properly the autoclaved probe assemblies can be stored dry for short periods of time lt 24 hr without loss of sensor longevity performance or sterility NOTE Before beginning probe insertion please become familiar with the Kleenpak Connector procedure outlined in Section 2 7 A E Probe Assembly Autoclave Tray for Probe Kits Figure 2 148 Thermo Scientific Single Use Bioreactor S U B 95 Section 2 5 Probe Assembly Probe Insertion 1 Insert probes into S U B BPC using autoclaved probe assemblies 50 1000 L a Place tubing clamps on all probe ports prior to attempting to connect probe assemblies this will prevent sterility loss in case errors are made during connection of Kleenpak Connectors Figure 2 149 Figure 2 149 b Position probe support plate onto outer support conta
116. ete Nevertheless readers should remain alert to the possibility of errors and omissions If any of the information in this manual seems incorrect contact Thermo Fisher Scientific for advice before proceeding This information including product specifications is subject to change without notice Because of this please take caution If extracting information from this User s Guide for use in other documentation as it will be updated with future revisions Please contact Thermo Scientific HyClone products technical support for the current revision MANUAL NUMBER 11 10 29 10 S U B User s Guide REV ECR ECN Date Description Thermo Scientific iI Thermo Scientific Section 1 Section 2 Section 3 Table of Contents Warnings and Safety 0 i cccccccccccceeeeeeeeseeseeeeeeeeeeeees i Single Use Bioreactor S U B Overview 1 1 Introduction to the Single Use Bioreactor ssesssseseeseseeseseressereseress 3 PD lid ware C Wat AC he S16 55s ak dies ces de recap seen esosuctenee a menue 5 L3 End User Supplied Components orestis iaa 7 1 4 BioProcess Container Characteristics esessesseseeeeseesesseresseresseresse 9 1 5 Additional System Components sssssssssessesessssesssressssessseresseresseres 12 1 6 Initial Installation Instructions ss ssssssseesessesessesessssessssresssresseresseres 13 ke Wnistallation and Sesa seach dea ientaeeeesBeteaeteaben 22 138 SUB Operanion Informationerne 24 1 9 S U B Operatin
117. ew the threaded cap covering the hollow pass through of the motor Figure 2 15 Figure 2 15 Mixing Assembly 15 Split shaft assembly 500 L system only Verify the two sections of the drive shaft are located in the drive shaft holders near the side of the outer support container The sections will be referred to as upper section with the drive shaft head and lower section with the square end Lubricate the threaded ends with a light coat of food grade anti seize with each use Insert lower section of drive shaft through hollow pass through of mixing assembly Figure 2 16 Once inserted slide latch pin from mixing assembly into shaft to prevent it from falling into the tube Figure 2 17 Assemble the upper and lower sections of the drive shaft by joining the segments and with a counterclockwise motion hand tighten the two sections together Figure 2 18 Remove latch pin Locate one wrench on the flat area in the lower drive shaft section another wrench on the upper section and tighten the connection using a counterclockwise rotation Figure 2 19 The shafts are reverse threaded to avoid loosening during operation CAUTION Do not over tighten a snug fit is sufficient Single Use Bioreactor S U B 43 Thermo Scientific Section 2 2 Single Use Bioreactor BPC Loading 50 500 L Volumes e Once the sections are secure return the wrenches to the tool holder Figure 2 17 Figure 2 18 Figure 2 19 Sing
118. g Instructions esssesseseeeesresesresssrerrereeserrerseresseres 31 Operating Information 2S AMD PATIO Procedure S mune gnana E EEEa 39 2 2 BPC Loading Instructions 50 100 250 and 500 L ou eee 40 2 3 BPC Loading Instructions 1000 L eeeseseeseseeeesssesseesssressressersss 49 2 4 BPC Loading Instructions 2000 L eseeesesssesessesssressresssressresssrsse 73 Diy VOWS SSM aa a EE E 95 ZO SaD E eoor E E ET 98 2 7 Pall Kleenpak Connector Instructions ssssssssseseeeseseesessresssrsseressees 100 Specifications 3 1 50 L S U B Data Sheet and Specifications ssseseseeesseseeesessereseeess 107 3 2 100 L S U B Data Sheet and Specifications sssseseseeseeesesseeeseeess 117 3 3 250 L S U B Data Sheet and Specifications ci ceeseeeeseeeeeseeeees 127 3 4 500 L S U B Data Sheet and Specifications sssseseseeseeesesseeseeess 137 3 5 1000 L S U B Data Sheet and Specifications 0 0 0 eeeeeeeeeeeeseeeeees 147 3 6 2000 L S U B Data Sheet and Specifications sssssssseesesssesseeeseeees 158 3 7 Standard BioProcess Containers with Disk Sparge Systems Data Sheet sisie noterne aiian 168 3 8 Condenser System Data Sheet and Specifications ceeeeseeeeeees 174 29 Load Celle Wie Data Sie a escyag ce eset iess vans E aS 178 3 10 S U B Vent Filter Heater Data Sheet cos sca seceh uate cten eiwet ate tsertiais 183 Thermo Scientific Section 4 Section 5 Appendix A Appendix B Appendix C Table of Con
119. g element is fully insulated with temperature range of 0 220 C Temperatures above series controller can be ordered as part of a complete molded silicone foam and is easily secured around the 60 C are not recommended for the filters used on the kit The vent filter heater system stands alone and filter by use of snap retainers This custom molded S U B The controller has programmable logic andis_ does not integrate with the control system of the heater creates a perfect fit that will fully encapsulate equipped with a low temperature alarm output Water S U B controller The system consists of the following the exhaust filters for consistent temperature protection is recommended for vent heater installation components Heater Controller and Power Cord Refer regulation The temperature controller is preset that may encounter wet conditions while in use to the vent filter heater user s manual included in the specifically for the HyClone Single Use Choose a power cord with a GFCI or for the utmost S U B ETP for further information Bioreactor U B at 50 C and has an adjustable protection a water resistant NEMA version of the 48 System Components and use Contoured foam insulator covers dome section to prevent heat loss ee Thermocouple embedded in heater mat Corrosion resistant stainless steel snaps for easy installation or removal Molded 1 2 silicone foam conforms to housing Water resistant NEMA 4X connectors
120. g media or supplements Please Note Not all options are available for all ports No customization of port type and location chamber dimensions or mixing assembly is possible For additional information please see the Selection Guides in the U B BPC Catalog Presentation as dry BPC systems Supplied flat packed Two polyethylene outer layers Description Product code Lot number Expiry date on outer packaging and shipping container Sterilization Irradiation 25 to 38 kGy inside outer packaging Shipping Container Durable cardboard carton Documentation Certificate of Analysis provided with each lot for each delivery Electric Resistive Heater SV50174 01 1000 L S U B Modbus Plus Load Cell Interface 240 VAC Includes 304 stainless steel outer support container with swivel SV50174 02 1000 L S U B Allen Bradley Load Cell Interface 240 VAC E e ieee heer aoa motor ae assembly with 3 piece split drive shart temperature controller SV50174 03 1000 LS U B Analog Output Load Cell Interface 240 VAC RTD sensor integrated resistive heating element three load cells SV50174 04 1000 L S U B Profibus Load Cell Interface 240 VAC with summing block display BPC pressure monitor probe shelf and SV50174 05 1000 L S U B DeviceNet Load Cell Interface 240 VAC standard tool set Water Jacket Includes 304 stainless steel outer support container with swivel caster platform variable speed agitation contr
121. gamma irradiation processes The S U B BPC does not operate as a closed system as it has both inlet and exhaust filters that are utilized to maintain an environment for cells to grow without concern for contamination However conditions can be encountered when gas inlet flow rate may exceed exhaust flow rate This may be encountered in the unlikely event of a pressure regulator failure on a gas feed or when excessive foaming creates conditions of vent blockage The S U B BPC is not rated as a pressure vessel gas pressure should not exceed 0 5 psi within the BPC Custom BPCs can be ordered with an optional disposable pressure transducer for monitoring the pressure within the S U B supplied standard with 1000 and 2000 L systems Each S U B is designed for a working volume range The minimum working volume and the rated working volume are listed in the specification tables provided in Section 3 The total volume listed includes the headspace needed for proper aeration and gas management Actual working volumes should not exceed the indicated rated working volumes In addition working volumes less than the minimums listed can result in damage to the S U B BPC and hardware malfunction The S U B is equipped with a bottom drain line that allows for liquid harvest by means of peristaltic pump Connection of the bottom drain line can be accomplished by use of a tubing welder or the aseptic connection of quick connect or fitting that is
122. ge 11 Base Addition 8 Exhaust Line 14 Open pipe gas sparge 10 Inoculum Addition Standard BPC Configuration CX5 14 Film For use with tubing welder and CPC Quick Connects Line Description Tubing Set ID X OD X Length End Treatment Pall Kleenpak aseptic 1 5 Probe Ports 5 1 2 12 7 mm Tube ports connectors KPCHT Series Female EEE a Thermo Well Adapter for 1 4 6 4 mm Diameter SterilEnz Pouch with small Volume Sample 1 8 3 2 mm ID X 1 4 6 4 mm OD C Flex X 24 60 cm injection site assembl 1 2 12 7 mm ID X 3 4 19 1 mm OD C Flex X 60 152 cm reduced to 3 8 9 5 mm ID X 5 8 15 9 mm OD C Flex X 12 30 cm splits to 1 4 6 4 mm ID X 7 16 11 1 mm OD C Flex X 12 30 cm reduced to 1 8 3 2 mm ID X 1 4 6 4 mm OD C Flex X 12 30 cm and 3 8 9 5 mm ID X 5 8 15 9 mm OD C Flex X 12 30 cm 1 2 12 7 mm ID X 3 4 19 1 mm OD C Flex X 4 10 cm splits to DETTE 1 2 12 7 mm ID X 3 4 19 1 mm OD C Flex X 10 25 cm and e a 1 2 12 7 mm ID X 3 4 19 1 mm OD C Flex X 10 25 cm Bottom Drain Harvest Plugged 3 8 MPC Insert Hydrophobic vent filter Overlay Gas Sparge 1 4 6 4 mm ID X 7 16 11 1 mm OD C Flex X 6 15 cm with Emflon Il Pressure Transducer a 1 4 6 4 mm ID X 7 16 11 1 mm OD C Flex X 84 213 cm reduced to Plugged Le 1 8 3 2 mm ID X 1 4 6 4 mm OD C Flex X 12 30 cm og
123. gement Actual working volumes should not exceed the indicated rated working volumes by greater than 10 percent In addition working volumes less than 50 percent of the rated volume can result in damage to the S U B BPC and hardware The standard S U B bag is designed with special spargers that produce very efficient mass transfer of oxygen and typically will require much less gas inflow than conventional spargers Gas inflow in fact only need be limited to prevent foam generation and excessive pressure within the BPC Gas flow rates supplied as overlay should also be reduced as much as possible or eliminated this will minimize both liquid evaporation and demand on the exhaust filter Ideally this will reduce the likelihood that gas inflows will exceed gas outflow of the system and reduce the occurrence of foam in the headspace that may plug the exhaust filter The exhaust vent filter used on the 50 L 1000 L S U B is a Pall KA3 series filter utilizing hydrophobic PVDF membranes These filters can be oriented in two directions 1 normal orientation with the flow arrow on the filter housing pointing away from the BPC and 2 reverse orientation with the flow atrow pointing toward the BPC In the normal orientation the filter has the maximum filter capacity however the side vents are positioned on the sterile side of the membrane Condensate removal in this orientation requires the use of a vent filter heater If the orientation is reversed t
124. gitation rate within the recommended range per Table 1 6 Allow speed to stabilize then make fine adjustments if necessary NOTE Agitation must be stopped when volume falls below minimum 1 2 of the rated volume otherwise damage to the hardware or disposable components may result a l After the agitation has been set turn on the temperature controller via the temperature controller keypad left button A flashing numeric display indicates the controller is off standby mode Adjust temperature set value using the up and down arrows located to the right The process temperature value is shown in red Single Use Bioreactor S U B 34 Section 1 9 5 U B Operating Instructions Temperature Control 1 Connection to an external temperature control unit TCU is made Water Jacket Systems via the large quick connect couplings located on the vessel jacket Ensure the inlet outlet ports are connected properly improper installation may result in poor heating cooling performance 2 Jacket should be purged of air any time the vessel jacket lines are reconnected 3 Temperature set points are controlled by the TCU Refer to the TCU manufacturer s guidelines for setup and operating instructions g g Drive Shaft Rotation Verify rotation of the drive shaft is operating in a counterclockwise motion when viewed from the top looking down The S U B is designed to mix in this direction only pH Probe Calibration In general the pH p
125. he filter capacity is reduced slightly however the side vents are now on the non sterile side allowing condensate to be removed with a syringe vent heater is optional The standard S U B BPC is supplied with the filter in the reversed orientation NOTE If normal orientation is desired this must be specified upon ordering and is considered a custom request It is important during operation the filter media be free of condensate in order to maximize the capacity of the vent filter To drain the condensate from the outer housing of the vent filter do the following 1 Turn the molded fitting of the lower side vent counterclockwise 2 Twist the side vent back and forth slightly to release the seal on the O ring inside the side vent 3 Attach a needleless luer lock syringe to the lower side vent and remove the condensate For users with more demanding applications an optional vent filter heater can be used see Section 5 3 for filter heater ordering information The exhaust vent filters used on the 2000 L S U B are Meissner UltraCap series filters utilizing hydrophobic PVDF membranes These filters are provided in normal orientation with the flow arrow on the filter housing pointing away from the BPC In the normal orientation the filter has the Single Use Bioreactor S U B 25 Sampling A Operating Pressure Thermo Scientific Section 1 8 S U B Operation Information maximum filter capacity No side vents
126. ilicone X 3 8 cm reduced to 1 4 6 4 mm ID X 7 16 11 1 mm OD C Flex X 4 10 cm reduced to 1 8 3 2 mm ID X 1 4 6 4 mm OD C Flex X 120 305 cm to 1 4 6 4 mm ID X 7 16 11 1 mm OD PharMed X 24 61 cm splits to 1 4 6 4 mm ID X 7 16 11 1 mm OD C Flex X 6 15 cm reduced to 1 8 3 2 mm ID X 1 4 6 4 mm OD C Flex X 12 30 cm and 1 4 6 4 mm ID X 7 16 11 1 mm OD C Flex X 18 46 cm Dip Tube 5 10 cm C Flex 3 8 9 5 mm ID X 5 8 15 9 mm OD C Flex X 120 305 cm to 3 8 9 5 mm ID X 5 8 15 9 mm OD PharMed X 24 61 cm split to 1 4 6 4 mm ID X 7 16 11 1 mm OD C Flex X 12 30 cm reduced to 1 8 3 2 mm ID X 1 4 6 4 mm OD C Flex X 12 30 cm and 3 8 9 5 mm ID X 5 8 15 9 mm OD C Flex X 12 30 cm Plugged Feed Antifoam 2 1 4 MPC Body Plugged Base Addition 3 8 MPC Body Dip Tube 6 15 cm C Flex 7 8 22 2 mm ID X 1 1 8 28 6 mm OD Silicone X 3 5 9cm reduced to 1 2 12 7 mm ID X 3 4 19 1 mm OD C Flex X 6 15 cm splits to 2 lines with 1 2 12 7 mm ID X 3 4 19 1 mm OD C Flex X 10 25 cm to 1 1 2 3 81 cm Triclamp Port i Connections for pH 7 Pall Kleenpak Aseptic DO probes 5 We sheet mam alte as Connector Female 12 Thermo Well Thermo Well Adapter for 1 4 6 4 mm Diameter SmartSite Valve Port small Volume Sample 1 8 3 2 mm ID X 1 4 6 4 mm OD C Flex X 24 61
127. ilter height above the S U B bag is make to order Must be compatible with a vent filter bracket option Choice of filters for inlets used to sterile filter incoming media or supplements Please Note Not all options are available for all ports No customization of port type and location chamber dimensions or mixing assembly is possible For additional information please see the Selection Guides in the U B BPC Catalog Presentation as dry BPC systems Supplied flat packed Two polyethylene outer layers Description Product code Lot number Expiry date on outer packaging and shipping container Irradiation 25 to 38 kGy inside outer packaging Durable cardboard carton Certificate of Analysis provided with each lot for each delivery Electric Resistive Heater Includes 304 stainless steel outer support container with swivel caster platform variable speed agitation controller motor drive as sembly with shaft PID temperature controller RTD sensor integrated resistive heating element probe shelf and standard tool set SV50200 02 500 L S U B 240 VAC Single Phase Water Jacket Includes 304 stainless steel outer support container with swivel caster platform variable speed agitation controller motor drive assembly with shaft RTD sensor integrated water jacket probe shelf and standard tool set SV50200 04 500 L S U B 240 VAC Single Phase Custom Hardware Options For stainless steel capabi
128. in BPC loading two operators may be required For drive shaft insertion a ladder or other means of elevation is required 1 Open both doors on bioreactor support container and reach inside to open clamp on bearing port receiver located below the motor Figure 2 32 Figure 2 33 If attached remove the probe shelf from the front of the tank cut out and store on the platform Figure 2 32 E Figure 2 33 Single Use Bioreactor S U B 49 Section 2 3 single Use Bioreactor BPC Loading 1000 L Electric Resistive Heater 2 Remove irradiated S U B BPC from protective double polybags Figure 2 34 Do not remove the polybags from the line sets at this stage as the BPC may become difficult to manage Do not allow the BPC or line sets to touch or drag on the floor Figure 2 34 3 Reach into or step inside the outer support container with the front face bearing port side of the BPC oriented towards the motor Figure 2 35 Figure 2 35 Thermo Scientific Single Use Bioreactor S U B 50 Section 2 3 single Use Bioreactor BPC Loading 1000 L Electric Resistive Heater 4 Place the top line sets still in polybags on top of the cross supports located on the top of the outer support container Figure 2 36 This will keep the BPC from being restricted during the air inflation step Figure 2 36 5 Load the BPC bearing port into the receiver Figure 2 37 and close the door and clamp Figure 2 38 Figure 237 Figure 2 38
129. in a smaller volume BPC as it is transferred by peristaltic pump into the S U B Floor scales can be used to measure the fluid content within the S U B This is accomplished by rolling the S U B onto the scale platform and leveling the S U B skid once in position Refer to Section 3 for skid dimensions for scale sizing Advanced Control Information External Data Logging The S U B hardware systems are designed to allow advanced users to and Control control all aspects of the operation of the bioreactor Contact technical support for Thermo Scientific HyClone products general integration guidance Thermo Scientific Single Use Bioreactor S U B 30 1 9 U B Operating Instructions Checkpoints prior to Media Fill Operating Conditions Thermo Scientific for Cell Culture Applications Section 1 9 5 U B Operating Instructions Verify the following before proceeding to liquid fill Load BPC into hardware following instructions provided in Section 2 All Kleenpak Connector port heavy duty clamps SV20664 01 or 02 are closed and located as close as possible to the S U B BPC Exhaust filter is upright and secured using the holder Clamp on drain tube is closed and located as close as possible to the S U B BPC Temperature RTD is completely seated in the thermo well and secured SS SS S Air filled bag is properly oriented in the outer support container and BPC bottom tabs are secured Operating parameter
130. iner This holds the probes at a 15 degree incline for best performance Figure 2 150 g i aj i a a i a r i a 1 Y aul Ei i i a he i t i 5 E w hh a r Z 4 jena F A n3 i a i h Hi J i A j S T Figure 2 150 c Connect Kleenpak Connector using protocol outlined by the manufacturer Section 2 7 d Ensure all four snaps click on each connector and the base and barrel of Kleenpak is fully seated e Remove tubing clamp on the individual probe port f Insert probe by collapsing bellows toward S U B Figure 2 151 Figure 2 151 g Probe is seated when bellow is fully collapsed A small amount of fluid may enter the bellow if the probe is loaded after media fill Thermo Scientific Single Use Bioreactor S U B 96 Section 2 5 Probe Assembly 2 Secure probe by placing connected probe assembly first into probe holder Figure 2 152 and then locking the holder into probe support shelf a Begin by placing the entire collapsed probe assembly in the probe holder starting with the Kleenpak Connector side being placed in the holder side with the short fingers b If necessary compress the probe bellows further to allow the probe assembly to be positioned and locked into the other end of the probe holder Figure 2 153 c Allow probe bellows to relax This secures the probe between the probe holder fingers d Align probe holder with alignment holes of the probe su
131. ing 9V50232 24 Thermoflex 900 TCU 240 VAC 50 or 60 Hz with necessary plumbing SV50241 01 Thermo Scientific Masterflex Peristaltic Pump with dual head l 115 VAC 50 or 60 Hz or 230 VAC 50 or 60 Hz Thermo Scientific Single Use Bioreactor S U B 196 Section 5 3 Ordering Information Accessories 5 3 Ordering Information Accesso ri CS Part Number Description SV50177 16 Elevated Single Filter Bracket Assembly for 50 100 and 250 L Elevated Filter Bracket Assembly 25 5 for dual filters tfits all sizes except 1000 L SV50177 17 jacketed S U B and 2000 L S U B SV50177 20 Elevated Dual Filter Bracket for 1000 L jacketed S U B SV50177 258 Dual Filter Bracket for 2000 L S U B tank when not using condenser system Accommodates two vent filter heater units SV50935 01 Mobile Stairs 2000 L S U B only S50191 01 09 Vent Filter Heater and Cord Options For use with exhaust filters when not using condenser system Thermo Scientific Single Use Bioreactor S U B 197 Section 5 4 Ordering Information Partial Spare Parts List 5 4 Ordering Information Partial Spare PartNumber Description Parts List SV50177 34 Drive shaft Assembly o0 L S U B One 1 Piece Integrated Design Delrin Anodized Aluminum SV50177 35 Drive shaft Assembly 250 L S U B One 1 Piece Integrated Design Delrin Anodized Aluminum V50177 36 Drive Shaft Assembly 500 L U B Two 2 Piece Integrated Design Delrin SST
132. ioProcess Containers with improved dual Thermo Scientific sparge capability Open pipe and e frit sparging systems are now HyClon e Si ngle Use included to provide a wider range Bioreactor S U B 500 L of operating conditions This document covers S U B systems with with agitator assembly sparger vent The HyClone S U B provides all the advantages of single use a maximum working volume of 500 L and the filter inlet and outlet ports plus ports for bioprocessing thani having to buy a complete new bioreactor new optimized standard S U B s with dual integration of sensor probes and line sets system The critical design parameters of the S U B such as height to sparge capability which provide a wider range The S U B BPC is supplied sterile and diameter ratios mixer design and location and typical control system of ka values better pH control and lower ready to use interfaces have been maintained foaming than the original disk sparger 3 Drive Shaft inserted into the S U B BPC The S U B BioProcess Container BPC is supplied sterilized through the mixing drive motor and locked into the disposable agitator assembly by irradiation and therefore does not require any facility hook ups for 4 Process Controls temperature not on sterilization or cleaning A key element to the single use design is the plastic polyethylene impeller with a bearing seal assembly linked to 1 Outer Support Container with a mixer water jacket
133. ir supply to the overlay can be turned off once the drive shaft has been inserted Thermo Scientific Single Use Bioreactor S U B 46 Section 2 2 single Use Bioreactor BPC Loading 50 500 L Volumes Final Installation Ste ps 19 Optional Wrap and secure vent filter heater on exhaust filter Connect heater to controller and verify it is plugged into an appropriate 120 or 240 VAC outlet then connect the power cord to the controller NOTE Controller is preset to 50 C 20 Secure exhaust vent filter on side mounted holder Figure 2 27A and 2 27B NOTE 500 L S U B bags and some custom S U B BPCs are supplied with dual exhaust vents Extended single and dual exhaust vent brackets are available see section 4 3 for ordering information Figure 2 27A Figure 2 27B 21 Position and close a bar clamp Part No SV20664 01 on the bottom drain line as close as possible to the BPC port Figure 2 28 Figure 2 28 Thermo Scientific Single Use Bioreactor S U B 4 Section 2 2 single Use Bioreactor BPC Loading 50 500 L Volumes 22 Insert RTD or selected temperature sensor into thermo well Figure 2 29 Figure 2 30 a Place small amount of glycerol 0 5 mL in well to aid in heat transfer b Sensor should be inserted until base of probe meets the mouth of the thermo well c Secure by twisting the luer lock collar if provided The thermo well will stretch slightly when RTD is seated Figure 2 29 Figure 2 30 23 Optio
134. irmly into assembly aid so that both membrane strips protrude through the opening and are easily accessible for removal Step e WARNING KLEENPAK CONNECTOR SHOULD STAY SECURELY IN THE ASSEMBLY AID WHEN PROPERLY INSTALLED A Remove Membrane Strips f Hold assembly aid in the palm of hand with Kleenpak Connector exposed outward and thumb is supporting the Kleenpak Connector in the unit Using the other hand firmly grasp both white peel away strips as close as possible to the body of the assembly aid to ensure a secure grip and pull them out simultaneously in a smooth continuous motion Ensure that the connector is perpendicular to the peel away strips WARNING DO NOT USE IF ONLY ONE WHITE PEEL AWAY STRIP IS REMOVED ACCIDENTALLY INSTEAD OF TWO THIS WILL AFFECT THE STERILITY OF THE PATHWAY A Thermo Scientific Single Use Bioreactor S U B 105 Thermo Scientific Section 2 7 Pall Kleenpak Connector Instructions Complete Flow Path Actuation g With Kleenpak Connector still secured in the assembly aid rotate and push the thumb rest of the male connector down towards the base of the barrel until they meet NOTE In order to establish a proper connection the plunger inside the male connector must be fully inserted into the female connector A h The thumb rest can be turned one full turn back to re align tubing as required WARNING VERIFY THAT THE PLUNGER HAS BEEN FULLY INSERTED INTO OPPOSING CONNECTION
135. is key to proceed through the sub block numbers until you find the choice of interest e Press select to toggle the values of the selected sub block e The S U B electrical schematic contains a table showing the sub blocks that have changed from the default settings Thermo Scientific Single Use Bioreactor S U B 204 Thermo Scientific Appendix C Mettler Toledo Load Cell Calibration Instructions Calibration Refer to technical manual section beginning on page 3 8 l 10 11 12 13 14 15 16 17 Access the setup menu by pressing select and print simultaneously Press print continually until sub block 1 4 is displayed Press print again until CAL X is displayed this is the calibration mode Press select to toggle the value from 0 to 1 this will enable the calibration mode Display will show E SCAL indicating you should now empty the scale Once the scale is empty press enter this will establish the scale zero Display will show 15 CAL and begin to count down to zero If the scale is not completely steady it may fault showing E SCALE If successful the display will show Add Ld Place test calibration weight on scale and press ENTER Display will show 00000 Now enter the value of test weight Once the value is entered press enter this will establish the scale span Display will show 15 CAL and begin to count down t
136. it with hardware Port sizes Limited customization possible as engineer to order with justification e g 1 ID port on harvest line Re arrangement of Limited customization possible e g moving sample tempwell port to oat roe SUES TTS Til ees a probe tube port or swapping overlay inlet line with supplement line Eee ee CE Ee OR Dag AT fE A A Dual sparger open pipe plus porous frit standard Can do an Sparger engineer to order for one or the other alone Make to order bags will be built around standard dual sparger chamber Din tube lines Limited customization possible Suggested use through 1 port so Length cannot interfere with impeller and shaft p this is engineer to order Otherwise must use ferrule approach Typical is 10 or shorter Overlay and Sparge Can use disposable capsule filter other than standard hydrophobic Line Filters vent filter with Emflon II Limited engineer to order customization possible such as additional Dependent on location in bag and compatibility media lines and vent filter lines Requires economic justification with hardware Vent Filters Standard is Pall Kleenpak 0 2 micron exhaust vent filter el filter heater configuration restricts ee Dig Extended filter height above the S U B bag is make to order Must be compatible with a vent filter bracket option OENE ET Choice of filters for inlets used to sterile filter incoming media or Supplement Inlets supplements Single Use Bioreactor S U B 193
137. itional ports second row of 4 for 50 L to 250 L S U Bs second row of 5 for 500 L and 1000 L U B s Pressure sensor Pendotech for 50 L to 250 L S U B comes standard on 500 L and 1000 L DO Finesse DO and pH PreSens Availability of various other disposable sensors currently pending Limited engineer to order customization possible such as additional Dependent on location in bag and compatibility media lines and vent filter lines Requires economic justification with hardware Limited customization possible as engineer to order with justification Dependent OI OC auOW IMSA ane iE Wit nanWalle e g 1 ID port on harvest line Limited customization possible e g moving sample tempwell port to tec anad a probe tube port or swapping overlay inlet line with supplement line Dependent an location in bag and it with hardware Dual sparger open pipe plus porous frit standard Can do an engineer to order for one or the other alone Make to order bags will be built around standard dual sparger chamber Limited customization possible Suggested use through 1 port so Length cannot interfere with impeller and shaft this is engineer to order Otherwise must use ferrule approach Typical is 10 or shorter Can use disposable capsule filter other than standard hydrophobic vent filter with Emflon Il Standard is Pall Kleenpak 0 2 micron exhaust vent filter Note Vent filter heater configuration restricts options Extended f
138. ive for Thermo Scientific HyClone products These items include all components that have part numbers beginning with the following two digits e SH e SV In the Americas and Asia In Europe Tel 1 435 792 8000 Tel 32 53 85 75 59 Toll free 1 800 HyClone Fax 32 53 85 74 31 Or 1 800 492 5663 perbio euromarketing thermofisher com Fax 1 435 792 8001 www thermo com perbio www thermoscientific com hyclone Info hyclone com Technical support for the Single Use Bioreactor is available through a variety of formats Some or all of the following may be appropriate depending on individual experience and circumstances Contact your Thermo Scientific HyClone sales representative for general product pricing availability delivery order information and product complaints Call 1 800 HyClone US or 32 53 85 75 59 EU for direct and immediate response to overall product questions and general product technical information Technical Support You can also contact our Tech Support by email at hyc bpctechsupport thermofisher com Appropriate technical support is available to assist in the initial setup and operation of each Single Use Bioreactor system Inquire at the time of purchase Training can be provided for startup and operation of the S U B Contact your Thermo Scientific HyClone Products sales representative Single Use Bioreactor S U B 200 Appendix A Electrical Receptacle Installation Instructions Appendix A
139. ively assemble and insert the drive shaft 3 Verify the drive shaft sections and tools are available 4 Prepare the hollow pass through by first removing the latch pin on the safety cover Figure 2 122 opening the safety cover and removing the threaded cap turn counterclockwise Figure 2 123 of the mixing assembly Figures 2 124 wa Figure 2 122 Figure 2 123 Cap H f J Drive Shaft Head Hollow Motor Drive Keyway Safety Cover Drive Shaft gt Figure 2 124 Mixing Assembly Thermo Scientific Single Use Bioreactor S U B 87 Section 2 4 single Use Bioreactor BPC Loading 2000 L Water Jacket Verify the four sections of the drive shaft are located in the drive shaft holders on the side of the outer support container The sections will be referred to as the upper section with the drive shaft head middle 2 each section with internal external quick connect on each end and lower section with the square end NOTE No lubrication is required with the quick connect assembly design A 5 First insert the lower section through the hollow pass through of the mixer drive Figure 2 125 Once inserted slide latch pin from mixing assembly into shaft to prevent it from falling into the impeller sleeve tube Figure 2 126 Figure 2 125 Figure 2 126 6 To connect two shafts together the button on the female side is depressed Figure 2 127 and the sleeve is slid back this will expose a red
140. just bioreactor controller to suite the volume of the S U B system Many different parameters can effect the ability of a bioreactor controller to effectively maintain a target setpoint during process control Modern controllers utilize computer algorithms to adjust targeted parameters the most common technique is that of tunable controller that uses variables of PID Tuning these PID values to the specific characteristics of the system dynamics will in most cases stabilize process parameters to an acceptable level It is recommended that S U B users consult the user manual of the particular bioreactor controller being used A grounding reference to the media can be created by using a grounding lead between the tank and the body of the stainless steel DO probe or to the stainless steel connector if present on the sample line of the S U B BPC Most traditional systems utilize a condenser or exhaust vent heater to help manage condensate is this necessary for the S U B Extensive testing has found that an exhaust heater or a condenser is not typically required under normal operating conditions for S U B s 500 L or smaller However more demanding applications may warrant an exhaust vent heater which is available from your sales rep I typically use level sensors to control volume and feed rate or supplement during a bioreactor run how would I do this with the S U B Use load cells or a scale to control volumes based upon weight
141. ket for use with Bracket for use with ES Z resistive heater S U B water jacketed S U B Wage Threaded Probe Attachment hi C cable Tie w o mp 3 i Bioreactor Probe STA U rd Molded Bellows Autoclave Tray B Noah Cover EE A p mua Additional information on autoclave tray Filter Bracket i e Fabricated from stainless steel Adapter for use Frit Sparge Line l 7 Cable Tie e Plastic carry handle for easy transport with resistive right out of the autoclave heater S U B i g ca T act e Positions probes on 15 incline for 2 i ter probe membrane longevit AE Pall ACD Connector PEIEE ply Jevy wi a e Will restrain probe bellows from ae collapsing during sterilization e Probe holder accommodates two probes SV50174 1000 L SH30774 05 SH30720 01 SV20664 03 SV50177 01 SV20750 01 SH30845 02 Necessary for regulation of gas pressure and flow rate control for DO and pH set points Autoclavable Probe 13 mm X 13 5PG thread w 195 235 mm insertion length Autoclavable Probe 13 mm X 13 5PG thread w 195 235 mm insertion length Tubing Welder Terumo or Wave Steam In Place SIP Sterilizer or Laminar Flow Hood Used for fluid transfer between line sets and containers Necessary for water jacket temperature control not provided S U B Hardware System standard S U B BPC standard Bioreactor Probe Assembly non sterile for use in autoclave Heavy Duty Tubing Clamp 10 pack Autoclave Tray for Autoclaving P
142. ks in tubing Plunger _ Step g Can twist while pushing the thumb rest of the male connector down towards the base of the barrel until they meet Single Use Bioreactor S U B 102 Thermo Scientific Section 2 7 Pall Kleenpak Connector Instructions Push the thumb rest of the male connector down towards the base of the barrel until they meet If the thumb rest has not already been turned counterclockwise to remove tubing kinks the thumb rest can be turned counterclockwise while it is being pushed towards the base of the barrel NOTE In order to establish a proper connection the plunger inside the male connector must be fully inserted into the female connector As a verification this normally occurs when the last or second to last rings on the plunger locks into the ratchet Minimally the second to last ring must be locked into the ratchet to ensure a proper connection Ihe thumb rest can be turned one full turn back to re align tubing as required if the second to last ring is locked into ratchet If the plunger has been pushed further than the second to last ring then it may not be possible to turn the thumb rest A Step h Start the fluid transfer Single Use Bioreactor S U B 103 Pall Kleenpak Connector Instructions for use with assembly aid Thermo Scientific Section 2 7 Pall Kleenpak Connector Instructions Receipt of the Equipment The connector assembly aid is provided non sterile and can b
143. ks or unintended clamps Verify the rear access door is closed with proper latch tension Remove the poly bag from the drain line set and verify redundant line clamps are in position Use a cable tie around the clamp to ensure the clamp cannot be accidentally opened Single Use Bioreactor S U B 91 Section 2 4 single Use Bioreactor BPC Loading 2000 L Water Jacket 4 Ensure the front access door is latched and align the five Kleenpak ports through the front access window Figure 2 137 This will be the lower cutout if hardware is supplied with two horizontal slots Position clamps as close as possible to the bag and close them on all tube ports Figure 2 138 5 Place clamps on subsurface lines as close to the port as possible this will eliminate media from filling these lines prior to use Figure 2 139 j H E ma r pa P Figure 2 137 Figure 2 138 Figure 2 139 6 Insert RTD or selected temperature sensor into the thermo well Figure 2 140 e Place a small amount of glycerol 0 5 mL in well to aid in heat transfer Glycerol Sigma G6279 e Sensor should be inserted until base of RTD meets the mouth of the thermo well e Secure by twisting the luer lock collar if provided The thermo well will stretch slightly when RTD is seated Figure 2 140 Connect batch to tank grounding cable to the stainless steel connector of the sample line Figure 2 141 NOTE Verify all port clamps are closed and located as close as
144. late NOTE The plug will be offset to the left or right if mounting to a 2 gang box with screws in opposite corners If this is the case then one of the holes in the plug will line up with the screw will need to be used in that hole Use other 2X 10X1 screws to plug holes not used 4X 6 32X 1 1 4 screws to mount adapter plate to TYP 2 gang FS FD box NOTE The plug will be offset to the left or right using 4X 10X1 screws Use other 2X 10X1 screws to plug holes not used Figure 3 Adapter Mounting Plate 7 Secure the electrical receptacle using four supplied screw fasteners Connect power back to the electrical circuit 9 Test the circuit with multi meter prior to making any connections to the electrical receptacle Thermo Scientific Single Use Bioreactor S U B 202 Appendix B Calibration of the AC Tech Variable Speed Drive Appendix B Calibration of the AC Tech Variable Speed Drive Calibration of the If during the verification of the mixer speed calibration it was determined AC Tech variable the displayed speed varied from the actual speed please follow these steps to calibrate the drive speed drive SCF 1 Press M on the speed controller menu SCM Models 2 If 000 appear the menu is locked To unlock the menu hold the up arrow key and select 225 then press the menu key 3 Use the arrow key to select P39 and then press M 4 Use the arrow keys to enter the correct va
145. le For additional information please see the Selection Guides in the U B BPC Catalog Presentation as dry BPC systems Supplied flat packed Two polyethylene outer layers Description Product code Lot number Expiry date on outer packaging and shipping container Sterilization Irradiation 25 to 40 kGy inside outer packaging Shipping Container Durable cardboard carton Documentation Certificate of Analysis provided with each lot for each delivery Water Jacket SV50223 11 2000 L S U B Modbus Plus Load Cell Interface 240 VAC Includes 304 stainless steel outer support container variable SV50223 12 2000 L S U B Allen Bradley Load Cell Interface 240 VAC speed agitation controller motor drive assembly with 4 segment SV50223 13 2000 L S U B Analog Output Load Cell Interface 240 VAC drive shaft RTD sensor integrated water jacket three load cells SV50223 14 2000 L S U B Profibus Load Cell Interface 240 VAC with summing block display BPC pressure monitor probe support SV50223 15 2000L S UB DeviceNet Load Cell Interface 240 VAC system and standard tool set Custom Hardware Options For stainless steel capabilities please contact your sales representative for HyClone products for more information peer Used to package the pH and DO probes not supplied for sterilization SH130720 01 Bioreactor Probe Assembly non sterile and to aseptically connect them to the S U B BPC See Figure 1
146. le Use Bioreactor S U B 44 Section 2 2 single Use Bioreactor BPC Loading 50 500 L Volumes Drive Shaft Insertion 16 Insert drive shaft through hollow pass through of mixing assembly in 7 the following manner Figures 2 20 2 23 50 500L Volumes e Use two hands to load drive shaft through top of mixing assembly a slight back and forth twisting motion will aid in insertion and avoid stretching of the impeller tubing e When approximately 2 3 5 08 cm of shaft remains twist slightly to engage impeller e When approximately 1 2 2 54 cm of shaft remains twist slightly to engage bearing assembly e When approximately 0 25 0 64 cm of shaft remains twist to align motor drive keyway with one of the four outer slots on the drive shaft head Figure 2 22 Figure 2 23 Thermo Scientific Single Use Bioreactor S U B 45 Section 2 2 single Use Bioreactor BPC Loading 50 500 L Volumes LZ ea couple drive shaft to motor drive Figures 2 24 2 26 Place threaded cap on hollow pass through and hand tighten clockwise Tighten cap by placing spanner wrench on hollow pass through and tighten cap using supplied torque wrench NOTE The torque wrench is a standard 3 8 square drive and it is calibrated at the factory at 150 in lbs Verify wrenches have been removed from system and placed in storage holders e Close safety access cover and insert latch pin Figure 2 25 Figure 2 26 18 The a
147. licone X 6 15 cm reduced to l 13 Direct Gas Sparge Check Valve and 1 4 6 4 mm ID X 7 16 11 1 mm OD C Flex X 48 122 cm Acro 30 Vent Filter Plugged SSV 3 4 mini TC Plugged 1 4 MPC Insert 2 on Standard BPC Configurations 500 and 1000 L For use with tubing welder CPC Quick Connect see Page 4 for line details Back Face 1 Overlay Gas Inlet 2 Media Inoculum 3 4 Antifoam 3 4 Feed 14 Direct Gas Sparge 6 Exhaust Filter Impeller Front Face 13 Bottom Drain Harvest 12 Thermo Well Small Volume Sample 7 11 Probe Ports 5 Standard BPC Configurations For use with tubing welder and CPC Quick Connects End Treatment Line Description Tubing Set ID X OD X Length With Quick Connects H30748 3 8 9 5 mm ID X 5 8 15 9 mm OD C Flex X 9 23 cm Pall Kleenpak Overlay Gas Inlet ele at e with inline disposable pressure transducer Gamma 0 2um 2 lines with 1 2 12 7 mm ID X 3 4 19 1 mm OD C Flex X 120 305 cm reduced to Media Inoculum 3 8 9 5 mm ID X 5 8 15 9 mm OD PharMed X 24 61 cm split to smartSite Valve Port Addition 1 4 6 4 mm ID X 7 16 11 1 mm OD C Flex X 6 15 cm reduced to 3 8 MPC Body 1 8 3 2 mm ID X 1 4 6 4 mm OD C Flex X 12 30 cm and 3 8 9 5 mm ID X 5 8 15 9 mm OD C Flex X 12 30 cm 1 2 12 7 mm ID X 3 4 19 1 mm OD S
148. lied by the user Water Jacket systems require a temperature control unit selected and supplied by the end user This User s Guide covers the setup operation maintenance and troubleshooting of all retrofit S U B systems in the following volumes 50 100 250 500 1000 and 2000 L Figure 1 1 Single Use Bioreactor Single Use Bioreactor S U B 4 1 2 Hardware Characteristics System Features Agitation Bioreactor Control System Exhaust Vent Filter Heater Condenser System Thermo Scientific Temperature Section 1 2 Hardware Characteristics The Single Use Bioreactor is designed for system mobility and easy disposable integration while utilizing a straightforward operator interface Hardware drawings and specifications tables specific to volume can be found in Section 3 The following sections describe the hardware components of the S U B Stirring speed is adjusted by using the keypad interface on the control panel The agitation control interface utilizes a digital display to indicate stirring speed in units of revolution per minute RPM Power is supplied to the motor by a two position power switch The up and down arrows on the agitation keypad adjust the stirring speed The Single Use Bioreactor is designed to integrate with existing bioreactor control systems in their numerous configurations The S U B control system manages the selected process parameters of agitation and temperature Parameters of
149. line sets of single use bioprocessing without having to buy a complete new i dual sparge capability which provide a wider The S U B BPC is supplied sterile and bioreactor system The critical design parameters of the S U B such as range of ka values better pH control and ready to use height to diameter ratios mixer design and location and typical control system Interfaces have been maintained The S U B BioProcess Container BPC is supplied sterilized by irradiation and therefore does not require any facility hook ups for Sterilization or cleaning A key element to the single use design Is the lower foaming than the original disk sparger 3 Drive Shaft inserted into the U B BPC through the mixing drive motor and locked into the disposable agitator assembly 4 Process Controls temperature not on iner wi water jacket and agitation control are plastic polyethylene impeller with a bearing seal assembly linked to 1 Outer Support Container with a mixer RN n f at pe baad aaa an external mixer drive drive complete with control unit and an Integrate Into the o U b but adaitiona electrical heater or water jacket pH or dissolved oxygen DO controls must 2 Single Use Bioreactor BPC complete be supplied by the user Rated Liquid Working Volume Minimum Liquid Working Volume Total Reactor Volume Liquid amp Gas BPC Chamber Diameter BPC Chamber Shoulder Height Liquid Height Rated Working Volume Overall Rea
150. ling Manifold with luer lock individual Available for aseptic luer connection or weld onto the sample line to SH30845 02 Sterile Sampling Manifold with luer lock 10 pack take sample sets 9V50177 65 Frit Sparge Line Support Recommended to maintain the frit sparge in a vertical position SV50177 16 Elevated Filter Bracket Needed to support an elevated vent filter Ny Probe Bioreactor Probe Autoclave Tray a Frit Sparge Line Assembly for Probe Kits p i tandia Support i Lis I hreaded Probe XN Tr Attachment A cabi Tie C AA Bioreactor Probe A Autoclave Tray Additional information on autoclave tray e Fabricated from stainless steel e Plastic carry handle for easy transport right out of the autoclave e Positions probes on 15 incline for greater probe membrane longevity Elevated Filter e Will restrain probe bellows from Se Pracket collapsing during sterilization oF e Probe holder accommodates two probes SH30845 02 Sterile Sampling Manifold with Luer Locks 10 pack 250 L with Resistive Heater The stainless steel outer support container contains the following features i Mixer motor 6 Drive shaft Zz Exhaust vent filter holder 2 Liquid sight window 3 Stainless steel 304 outer support 8 Standard tool set container g Probe access window 4 Bearing port receiver with clamp 10 Control panel D Mixing assembly with shield 11 Probe holder Probe shelf Platform with swivel casters an
151. lities please contact your sales representative for HyClone products for more information oe Used to package the pH and DO probes not supplied for sterilization a eu PIO aN PLO nese ely Sas e ey and to aseptically connect them to the S U B BPC See Figure 1 SV20664 01 Heavy Duty Tubing Clamp SV20664 02 Heavy Duty Tubing Clamp 4 pack of SV20664 01 SV20664 03 Heavy Duty Tubing Clamp 10 pack of SV20664 01 Reusable tubing clamp used on probe ports when connecting probe assemblies SV20750 01 S U B Temperature Sample Port Used for RTD calibration validation Support tray which allows the probes to be safely autoclaved See SV50177 01 Autoclave Tray for Probe Kits stainless steel Foue SH30845 01 Sterile Sampling Manifold with luer lock individual Available for aseptic luer connection or weld onto the sample line to SH30845 02 Sterile Sampling Manifold with luer lock 10 pack take sample sets 5V50177 65 Frit Sparge Line Support Recommended to maintain the frit sparge in a vertical position 9V50177 17 Elevated Dual Filter Bracket Needed to support elevated dual vent filters Bioreactor Probe Assembly Threaded Probe os Attachment C y Cable Tie Molded Bellows Cover C z Cable Tie a Sy F Ne Pall ACD Connector ey Probe Autoclave Tray l for Probe Kits M i Handle Bioreactor Probe A by Autoclave Tray Additional information on autoclave tray e Fabricated
152. locked into the disposable agitator assembly 4 Process Controls temperature not on plastic polyethylene impeller with a bearing seal assembly linked to 1 Outer Support Container with a mixer water Jacket and agitation control are an external mixer drive drive complete with control unit and an integrated into the U B but additional electrical heater or water jacket pH or dissolved oxygen DQ controls must 2 Single Use Bioreactor BPC complete be supplied by the user Rated Liquid Working Volume height diameter ratio Impeller Diameter pH amp DO Probe Autoclavable Type Applisens Broadley James Metler Toledo insertion length x 13 5 PG thread Electrical Power Supply Requirement 208 240 VAC single voltage phase amp 30 A 30 A 46 116 8 cm 60 153 4 cm 90 228 6 cm _S U B Recommended Operating Parameters Operating Temperature Range Motor Speed Volume Range Maximum Bag Pressure 0 5 psi 0 03 bar Continuous Operating Time 21 days Mixing time at nominal volume only 1000 L Water Jacket S U B nna Part of Thermo Fisher Scientific pee Ge S os A DA E A a E O N 0J m re Standard BPC Configuration SH30774 05 For use with tubing welder and CPC Quick Connects see page 3 for line details 1 5 Probe Ports 5 Impeller 7 Bottom Drain Harvest Front Face Back Face W 9 Overlay Gas Sparge m f y fe 12 13 Feed Lines 15 Porous frit gas spar
153. lting in excessive internal pressure within the confines of the S U B This may cause product failure and impending burst of the S U B BPC Sym ptom Noise emitted from the BPC mixer assembly Remedy No action required The bearing port assembly supplied with the S U B is an important component in maintaining a sterile environment during cell growth The special seals used in the S U B may generate some noise during operation particularly after the first day of operation This noise may vary in intensity and frequency but generally has no significant effect upon performance or overall durability of the S U B BPC during the intended life of the product Thermo Scientific Single Use Bioreactor S U B 189 Section 4 Troubleshooting Symptom We forgot to introduce the pH and DO probes prior to media fill can we still make a sterile connection under these conditions Remedy Yes as long as the clamps were closed on the Kleenpak Connector probe ports before liquid fill e The Kleenpak connectors must be dry to make the connection of the probe assemblies When media is already present in the S U B follow the probe insertion procedures as outlined in Section 2 5 Probe Assembly Some fluid may enter the bellows when the probe is inserted into a BPC already filled with media Sym ptom I am not familiar with the use of the Pall Kleenpak Connectors and am concerned about making connections What can I do to ensure a successful con
154. lue listed in the Table 3 1 then press M NOTE The value may need to be adjusted slightly to calibrate exactly S U B VFD Parameters for RPM Calibration 1 rpm 50 L SCF SCM P39 178 Motor rating of 1780 rom 60 Hz with gear models reduction of 10 1 1780 10 178 100 L SCF SCM P39 178 Motor rating of 1780 rom 60 Hz with gear models reduction of 10 1 1780 12 5 178 250 L SCF SCM P39 147 Motor rating of 1780 rom 60 Hz with gear models reduction of 12 5 1 1780 12 5 142 500 L SCF SCM P39 356 Motor rating of 1780 rom 60 Hz with gear models reduction of 5 1 1780 5 1 356 1000 L SCF SCM P39 119 Motor rating of 1780 rom 60 Hz with gear models reduction of 15 1 1780 15 119 2000 L SCF SCM P39 119 Motor rating of 1780 rom 60 Hz with gear models reduction of 15 1 1780 15 119 Table 3 1 VFD Parameters 5 The mixer output should now be scaled properly for the correct RPM displayed NOTE For users who currently operate the S U B and have not previously verified the calibration of the system the current calibration of the system should be verified so an approximate operating RPM for any previous bioreactor runs can be documented and compared to the newly calibrated system Users can then verify the parameters in the above table and change them if necessary To confirm that the parameter change provides a calibration accuracy of 1 RPM conduct a final check of the calibration refe
155. m Recommended for 2000 L S U B Optional on smaller systems e Load cells Standard on 1000 and 2000 L S U B s Optional on smaller sizes Consists of three load cells summing block wiring and display with choice of several data interfaces Single Use Bioreactor S U B 12 1 6 Initial Installation Instructions Section 1 6 Initial Installation Instructions Hardware Shipment The Single Use Bioreactor hardware is shipped directly from the and Setup manufacturer and will arrive crated For unpacking instructions and detailed contents of the crate please refer to the instructions specific to the p p S U B volume Be sure to follow the unpacking instructions provided and retain all packaging materials 50 L 100 L 250 L and The S U B hardware should arrive with the following items 500 L Hardware Uncrating a aie one ae Outer support container platform tank and control panel Drive shaft RTD probe shelf and brackets 4 and standard tool set spanner wrench and torque wrench ETP located on CD provided with hardware Remove crate lid first Remove side wall s Remove the tie down straps Remove any all blocks on crate floor Remove S U B carefully using forklift manually by lifting via the bottom of the cart frame not the casters Ensure no damage occurred during shipping Contact your Sales Representative immediately if damage has occurred 1000 L and 2000 L Detailed instructions for crating uncrating an
156. m could fail causing personal injury or damage to equipment DO NOT leave the BPC unattended while inflating a 10 Begin air inflation through the overlay gas line 11 While the BPC is inflating attach the incoming gas supply to the sparger gas inlet line NOTE Air pressure to the sparger on the S U B BPC should not exceed 8 psi Also while a sparge line check valve is provided for each sparge line it is not uncommon for some fluid to bypass check valves during typical use Elevating the filter to ensure that it is not at the low point of the sparge line will reduce the chance that the filter is exposed to liquid a 12 Tare the load cell display before proceeding 13 Using the four stretch hooks provided secure the BPC by attaching the hooks to the hanging tabs on the bottom of the BPC Figure 2 43 Figure 2 44 Verify that the sparge filter and sparge remain in position while attaching the stretch hooks It is recommended that users secure the hanging tabs on the front BPC panel first This way the door will not be an obstruction when connecting the last set of stretch hooks Figure 2 43 Figure 2 44 Thermo Scientific Single Use Bioreactor S U B 53 Section 2 3 single Use Bioreactor BPC Loading 1000 L Electric Resistive Heater 14 While BPC begins to fill with air remove the protective packaging from the exhaust vent filters Figure 2 45 Figure 2 45 15 Air fill of the S U B BPC takes approximately 1
157. mo Scientific Section 2 4 single Use Bioreactor BPC Loading 2000 L Water Jacket 9 The first operator loads the peristaltic tubing into the pump verifying there is sufficient slack at each end of the pump tubing aligns tubing in the pump channel closes the pump ramp Figure 2 117 10 The pump is then started pushing the red button Figure 2 118 Operators verify that both the pump and chiller are enabled and running at the proper settings we recommend that the pump setting be 12 30 RPM and the chiller 5 C Specified pumping system is qualified to run continuously wet or dry beyond 21 days v Figure 2 117 Figure 2 118 11 After setup the operators should verify the following e The elbow fittings on the inlet and outlet of the condenser saddle bag are straight and level e The gas inlet line and the condensate line are not twisted pinched or obstructed e Low spots in the gas inlet line should be avoided Adjust the lines to avoid condensation pooling e The pump union is loose on both ends of the pump and running smoothly in the peristaltic rollers Specific performance questions can be answered by reviewing the Disposable Condenser portion of the S U B Validation Guide or contacting technical support Single Use Bioreactor S U B 85 Section 2 4 single Use Bioreactor BPC Loading 2000 L Water Jacket Exhaust Vent Heater 1 A single operator can safely set up the exhaust vents and heaters in op
158. mount not shown Load cells 3 Load cell summing block Jacket quick connect couplings Data Sheet Outer Support Container units in inches mm 1000 L S U B Electric Resistive Heater 1000 L S U B Water Jacket 1004 98 aa RIG gar 301 20 LA SOFT Cay FI B 7 ff 100745 eE pre 7a rak aid i MONp aa TANK IG MET lisa 745 20 par 1318 87 OD TANK 51 7i osa a4 i oar ai ID TANK 2a BO 1a 1431 04 1973 89 oo E 54 I 43 47 1 a4 4230 li wE 132 57 5 y 2010 Thermo Fisher Scientific Inc All rights reserved SmartSite is a trademark of ALARIS Medical Systems Inc C Flex and PharMed are registered HyClone Laboratories inc Logan UT USA is 0 Certified trademarks of Saint Gobain PharmaPure is a registered trademark of PharmaPure Tubing Clave is a registered trademark of ICU Medical Pall Kleenpak and Emflon are registered trademarks of Pall Corporation Lynx is a registered trademark of Millipore SterilEnz is a registered trademark of PAW Bioscience Products INC All other trademarks are the property of Thermo Fisher Scientific Inc and its subsidiaries Specifications terms and pricing are subject to change Not all products are available in all countries Please consult your local sales representative for details Cell Culture amp BioProcessing 925 West 1800 South In Americas Asia In Europe www thermoscientific com hyclon
159. mperature the S U B is ready for inoculation Connect the inoculum addition line set to the seed culture vessel equipped with the proper connectors tubing and transfer the inoculum into the S U B Typically this is done with the tubing connection process aseptic luer lock connection or tube welding and peristaltic pump Pump the desired volume of seed cells into the S U B NOTE For shear sensitive cultures cells can be introduced by manipulating the addition port to direct the inoculum down the interior wall of the BPC and into the bulk fluid reducing the shear on the cells Custom line sets can be supplied with dip tubes which shorten the distance between the point of inoculum introduction and the bulk fluid level Verify the following once or twice daily during the culture run Visual verification of sparging rising bubbles through the sparge access window Verification of process parameters such as temperature and agitation are at setpoint Bag is not operating under pressure Cap is tight on drive shaft Temperature RTD is completely seated and secured Se os OC That no condensate accumulates in exhaust filter housing If condensate has accumulated then this indicates that use of a filter heater or a Condenser System is required NOTE Noise may be emitted from the mixer assembly during operation This noise may vary in intensity and frequency but generally has no significant effect upon performance or overall durability
160. mplete assembly procedures and technical specifications for Kleenpak connectors are contained in Pall publication USD 2233d e Please follow carefully the instruction below Pall Kleenpak Connector Standard Instructions a Lift and pull tab of protective caps to remove caps from connectors Step a Prolectne caps E h crf deni Thermo Scientific Single Use Bioreactor S U B 100 Section 2 7 Pall Kleenpak Connector Instructions b Hold the barrel of the larger male connector above the base Align the smaller female connector with the larger male connector so that both folded white peel away strips are facing the same direction as they exit out the flat sides of the connectors Step b Female Connector Z _Anti actuation E mng Flat side ii whore beth tedd ver peel maoy Sinipa w Male Connector NOTE Rounded sides must be in alignment and the white peel away strip must remain folded If the connectors are not aligned properly the connection cannot be made The peel away strips must remain folded A c Once aligned correctly press the two connectors together firmly until both sets of locking clips snap together tightly Step c P _ Locking clips reel ses Locking clips W F d While holding the Kleenpak Connector securely in hand pull the anti actuation ring outward Ramowe thie ambi nine from tha male connector Step d B Thermo Scientific Single Use Bi
161. n 3 4 3 500 L Hardware Specifications Water Jacket fanbase Impeler Scing Geele temeer O OOOO O with Exterior Insulation Shield 5 to 40 0 1 C 104 0 2 F Asyawoay 10373983Y ETEL siajaweleg U018 Y 101 u09 dway sauiejuo4 yoddns TIENER Single Use Bioreactor S U B 145 Section 3 4 4 500 L Control Panel Layout 3 4 4 500 L Control Panel Layout 500 L Control Panel Electric Resistive Heater Main Power Source 16 Pin Communication Quick Connect speed Control Key Pad 7 Motor Controller On Off Switch External Heater Temperature Controller Relay Input Main Switch Heater Cable Motor Cable 500 L Control Panel Water Jacket Speed Control Key Pad Main Power Source m 16 Pin Communication Quick Connect Motor Controller On Off Switch Emergency Stop Main Switch Motor Cable Thermo Scientific Single Use Bioreactor S U B 146 3 5 1000 L Single Use Thermo Scientific Bioreactor 3 5 1 22 ID 3 5 4 Section 3 5 1000 L Single Use Bioreactor 1000 L S U B Data Sheet 1000 L Technical Specifications Electric Resistive Heater 1000 L Technical Specifications Water Jacket 1000 L Control Panel Layout Single Use Bioreactor S U B 147 The Thermo Scientific HyClone Single Use Bioreactor S U B is the leading single use alternative to conventional stirred tank bioreactors for a
162. nal Connect pressure transducer to user supplied monitoring device 24 Continue to Section 2 5 for probe insertion instructions 25 Close the bottom access hatch 500 L only The proper latch tension can be obtained by a combination of feel and visual inspection When closing the latch the handle should begin to provide resistance to closing when the leading edge of the safety pin pass through of the latch handle aligns with the outside edge of the latch base Figure 2 31 NOTE When the latch is under tensioned the safety pin pass through of the latch handle will be covered within the latch base and the handle will close very easily If the latch is over tensioned the handle will be excessively difficult to close 26 The access doors must be closed and fully latched prior to filling the system with liquid Outside Edge of Latch Base safety Pin Pass Through of Latch set Latch Retainer safety Pin Pass Through of Latch Handle Figure 2 31 Thermo Scientific Single Use Bioreactor S U B 48 2 3 Single Use Bioreactor BPC Loading for 1000 L Electric Resistive Heater Thermo Scientific Section 2 3 single Use Bioreactor BPC Loading 1000 L Electric Resistive Heater Before beginning verify that the outer support container is stationary with casters locked into place BPC loading may require operators to step inside the bioreactor and the unit must be stationary for the safety of both operator and equipment For ease
163. nection using this system Remedy Familiarize yourself with the Pall Kleenpak Connector instructions found in Section 2 7 before beginning e When a connection is being made visually evaluate the status of the four locking external clips and verify they are tightly secured the snap should be audible for all four clips when pressing the connectors together Always make sure the four locking clips are fully engaged for the male female connection before removing the paper strips e Acommon cause for a leaky Kleenpak is an error in the final step of seating the tapered barrels of the male female connector There are a series of concentric rings inside the male connector 0 3 in front of the black O ring Visually verify that the four internal clips are on the last set of rings Using both hands place connector flanges between index fingers and thumbs and squeeze until properly seated Thermo Scientific Single Use Bioreactor S U B 190 Thermo Scientific Section 4 Troubleshooting Sym ptom We are not achieving the cell growth we expected in the S U B while running under our normal bioreactor agitation and sparging rates What should we do Remedy Reduce agitation and sparging rates Often low cell viability and cell growth can be attributed to excessive sparging or agitation It is recommended that users reduce their sparge rate compared to what they might use in a conventional bioreactor Gas flow rates supplied as overlay shoul
164. ng from the S U B The liquid condensate that is stripped from the exhaust gasses is then pumped back into the S U B chamber creating a sterile loop and significantly reducing liquid loss due to evaporation The condenser plate is chilled by a closed bath recirculating chiller which has sufficient capacity to cool two condenser plates simultaneously Figure 2 107 shows a functional diagram of the condenser system Exhaust Line l Filter Straps i z Filter Bracket From S U B _ Assembly Condenser Post Assembly Exhaust Vent ia 3 i reveal Plate Filters F amp _ Post Receivers Condenser Return Line Back to S U B Dual Headed Condenser Bag Gas Outlet Port Condenser Bag Gas Inlet Port aA Dual Chamber E Closed Bath Assembly Condenser Bag Gripping Recirculating j Tabs Chiller Condenser Bag i Liquid Drain Ports Alignment Holes Condenser Disposables Condenser Hardware Condenser System Figure 2 107 Condenser System Overview Thermo Scientific Single Use Bioreactor S U B 78 Section 2 4 single Use Bioreactor BPC Loading 2000 L Water Jacket Guidance for when to use the Condenser System 2000 L S U B BPC with Large 10 hydrophobic PVDF filters with a nominal 0 2 micron pore Single Use Condenser size were specified in order to increase the available surface area for off System gassing In conjunction the standard 2000 L S U B is designed for use with a single
165. nimal and insect cell culture The current range includes units with maximum working volumes of 50 100 250 500 1000 and 2000 L The S U B is now available with new optimized standard BioProcess Containers with improved dual Thermo Scientific sparge capability Open pipe and frit sparging systems are now HyClon e Si ngle Use included to provide a wider range of operating conditions Bioreactor S U B 1000 L Po This document covers S U B systems with with agitator assembly sparger vent a maximum working volume of 1000 L and filter inlet and outlet ports plus ports for The HyClone S U B provides all the advantages of single use the new optimized standard U B s with dual integration of sensor probes and line sets bioprocessing without having to buy a complete new bioreactor oe i sparge capability which provide a wider range The S U B BPC is supplied sterile and system The critical design parameters of the S U B such as height to diameter ratios mixer design and location and typical control system of KA values better pH control and lower ey ee d YP y foaming than the original disk sparger 3 Drive Shaft inserted into the S U B BPC interfaces have been maintained The S U B BioProcess Container BPC is supplied sterilized by irradiation and therefore does not require any facility hook ups for Sterilization or cleaning A key element to the single use design is the through the mixing drive motor and
166. nt strong consideration when performing batch runs beyond 10 days Results will vary however it is strongly recommended that end users select a vent filter configuration providing reserve capacity where possible For example dual vent configurations can be used independently with the second filter serving as a redundant backup providing a quick reserve in case issues arise in process Off Gassing Management Guidelines for Single Use Bioreactor Condenser System vs Exhaust Vent Heaters Conditions 37 C batch temperature no foam in exhaust stream O l tez S N N D wh _ o T m Run Duration days Condenser System 2 X 10 vents Vent Heater System 2 X 10 vents Vent Heater System 2 X 3 vents Vent Heater System 1 X 3 vents Graph 2 2 Thermo Scientific Single Use Bioreactor S U B 82 Section 2 4 single Use Bioreactor BPC Loading 2000 L Water Jacket Condenser System Two operators are required to safely setup the disposable exhaust system Functional Overview Setup time is typically 2 3 min cont 1 The first operator located at the elevated position should remove the condenser BPC carefully from the poly bag packaging and then lower the assembly directed in a vents first orientation to the second operator located at ground level standing to the rear but between the condenser cart and the S U B Figure 2 108 2 The first operator at the upper
167. ntific HyClone Single Use 1000 L S U B Bioreactor S U B and the Thermo Scientific HyClone Single Use Mixer S U M Load Cell systems include three cells summing block display and wiring They can be purchased either at the time of equipment sale or as retro fit kits for existing S U B and S U M units Units can FLEXMOUNT Load Cell Load Cell Specifications Weigh Module Parameter Unit of Measure Specification 50 amp 100 LS U B 250 L S U B 500 amp 1000 L S U B Model Number ln Te 0958 FLEXMOUNT Rated Capacity Pkg ty 113 250 220 500 550 1250 1100 2500 Load Limit Safe Stainless Steel kN Ib 1 7 375 3 3 750 8 3 1875 16 6 3750 KN Ib 1 1 250 2 2 500 4 1 920 8 2 1840 Maximum Horizontal Force Longitudinal kN Ib 1 1 250 2 2 500 5 6 1250 11 2 2500 TOE Maximum Top Plate Travel Longitudinal 3 01 0 12 Maximum Uplift Force OO kN Ib 22 2 5000 Weight including load cell nominal ee kg Ib 7 15 5 Material a a 304 Stainless Steel 1 Maximum horizontal force that can be applied to the top plate in a direction transverse to the longitudinal axis of the load cell Applies to fixed pin module and semi float module assembled in standard radial mounting position 2 Maximum horizontal force that can be applied to the top plate in a direction parallel to the longitudinal axis of the load cell Applies to fixed pin module and semi float module assembled in nonstandard tangential mounting p
168. o zero Display will show CAL d meaning calibration is done Exit setup mode if task is complete by pressing CLEAR Display will show CaL OFF Then press PRINT The Panther Display will return to normal operating mode Single Use Bioreactor S U B 205 2010 Thermo Fisher Scientific All rights reserved Cell Culture amp BioProcessing 925 West 1800 South In Americas Asia In Europe www thermoscientific com hyclone Logan UT 435 792 8000 32 53 85 71 80 84321 435 792 8001 fax 32 53 85 74 31 fax
169. of the S U B BPC during the intended life of the product a 1 Connect bottom drain tubing set to intended transfer line 2 Open clamp positioned at the bottom drain port 3 Begin drain by use of peristaltic pump 4 When approximately 3 5 liters remain in the S U B BPC lift the S U B BPC at the top hanging tabs located opposite of the bottom drain this will pool media towards drain 5 Hold bottom drain line near floor while lifting exhaust filter side of S U B BPC to facilitate the drain of the final liter of harvest media Single Use Bioreactor S U B 36 S U B BPC Disposal S U B Shutdown Thermo Scientific Preparation for Next Run Section 1 9 5 U B Operating Instructions Once the S U B is drained the drive shaft can be removed and stored by reversing the steps used during assembly The U B BPC can then be removed from the outer support container Filters can be removed and integrity tested as needed as per user s standard procedures All product contact materials related to the S U B can be disposed of in an appropriate waste container or incinerator 1 Once run is complete verify motor agitation is off and turn off the power to the outer support container by switching off the main power disconnect 2 Ifthe S U B hardware has come in contact with caustic materials during the course of a run rinse affected areas with a light water rinse followed by normal routine cleaning see Preparation for Next
170. oller motor drive assembly with 3 piece split drive shaft RTD sensor integrated water jacket three load cells with summing block display BPC pressure monitor probe shelf and standard tool set Custom Hardware Options For stainless steel capabilities please contact your sales representative for HyClone products for more information ae Used to package the pH and DO probes not supplied for sterilization SH130720 01 Bioreactor Probe Assembly non sterile and to aseptically connect them to the S U B BPC See Figure 1 eri clamp used on probe ports when connecting probe SV20750 01 SV50177 01 Autoclave Tray for Probe Kits stainless steel o tray which allows the probes to be safely autoclaved See SH30845 01 Sterile Sampling Manifold with luer lock individual _ Available for aseptic luer connection or weld onto the sample line to SH30845 02 Sterile Sampling Manifold with luer lock 10 pack take sample sets SV50177 65 Frit Sparge Line Support Recommended to maintain the frit sparge in a vertical position SV50177 17 Elevated Dual Filter Bracket resistive blanket S U B Needed to support elevated dual vent filters SV50177 21 Adapter required for SV50177 17 D SV50177 20 E ane Bracket jacketed 9 U B Needed to support elevated dual vent filters No adapter required k Probe Bioreactor Probe Autoclave Tray A a Elevated Dual Filter Elevated Dual Filter Assembly for Probe Kits a oe Handie Brac
171. om 1 8 to 3 16 Once the proper clearance has been achieved retighten the jam nut lean IAS EATA TOP PLATE P CENTERING WASHER amp NUT SHOWN IN WEIGHING POSITION BASE PLATE SECTION A Figure 1 13 Unlocking Load Cells At this point the S U B hardware is ready to be prepared for a cell culture run CAUTION Do not move unit especially when filled while load cells are unlocked as this can damage the load cells For load cell calibration refer to Appendix C Single Use Bioreactor S U B 19 Section 1 6 Initial Installation Instructions ph 2000 L Outer Support Container Preparation Using a flat head screwdriver open the control box and locate the breakers for the pressure sensor continuous power outlets non E stoppable 2 continuous power outlets E stoppable 2 Figure 1 14 These breakers should be in the ON position during operation blue indicator pressed in For electrical schematics please refer to the ETP which is located on CD provided with hardware 2 Verify position of the three way motor controller switch it should be in the middle position for reference the middle position is to enable the speed control keypad 3 Close the control box and lock the panel using a screwdriver before continuing 4 For S U B hardware units purchased with factory installed load cells the load cells are shipped in the locked position centering washer Load Cell Preparation and nut ar
172. on 2 4 single Use Bioreactor BPC Loading 2000 L Water Jacket 12 Using two hands carefully guide the completed drive shaft into the BPC using a slight back and forth twisting motion or a counter clockwise rotation Figure 2 132 e When 2 3 5 08 cm of shaft remains twist slightly to engage the impeller e When 1 2 2 54 cm of shaft remains twist slightly to engage bearing assembly Figure 2 133 e When 0 25 0 64 cm of shaft remains twist to align motor drive keyway with one of the four outer slots on the drive shaft head Figure 2 134 13 Ensure the head is fully seated before directly coupling the drive shaft to the motor by placing the threaded cap back on the hollow pass through and tightening it NOTE The cap should be easy to install when the drive shaft head is fully engaged in the hollow pass through Otherwise repeat all steps of 13 before installing cap A Figure 2 133 Figure 2 13 14 Secure cap by placing spanner wrench on hollow pass through and tightening using the supplied torque wrench Figure 2 135 NOTE The torque wrench is a standard 3 8 square drive and it is calibrated at the factory at 150 in lbs A 15 Verify wrenches have been removed from the system and returned to the storage holders 16 Close safety access cover and insert latch pin Thermo Scientific Single Use Bioreactor S U B 90 Securing Access Doors Final BPC Installation Thermo Scientific Steps
173. oper tension is obtained by adjusting the threaded latch pin Tension of the latch is adjusted by varying the position of the pin on the threaded shank Figure 2 59 Figure 2 59 Thermo Scientific Single Use Bioreactor S U B 59 Section 2 3 single Use Bioreactor BPC Loading 1000 L Electric Resistive Heater 27 The proper latch tension can be obtained by a combination of feel and visual inspection When closing the latch the handle should begin to provide resistance to closing when the leading edge of the safety pin pass through of the latch handle aligns with the outside edge of the latch base Figure 2 60 NOTE When the latch is under tensioned the safety pin pass through of the latch handle will be covered within the latch base and the handle will close very easily If the latch is over tensioned the handle will be excessively difficult to close Outside Edge of Latch Base safety Pin Pass Through of Latch Base Latch Retainer safety Pin Pass Through of Latch Handle Figure 2 60 28 Once the two middle latches are closed the top latch 1 and bottom 2 30 31 Ti 33 34 OD Thermo Scientific latches latch 4 can be closed using the same tensioning procedures Both the left and right access doors must be closed and fully latched prior to proceeding to the final latch tensioning procedures The final procedure that follows must be performed one latch at a time Release latch 2 notice tha
174. order bags will be built around standard dual sparger chamber Limited customization possible Suggested use through 1 port so Length cannot interfere with impeller and shaft this is engineer to order Otherwise must use ferrule approach Typical is 10 or shorter Can use disposable capsule filter other than standard hydrophobic vent filter with Emflon Il standard is Pall Kleenpak 0 2 micron exhaust vent filter Note Vent filter heater configuration restricts options Heal Extended filter height above the S U B bag is make to order Must be compatible with a vent filter bracket option Choice of filters for inlets used to sterile filter incoming media or supplements Please Note Not all options are available for all ports No customization of port type and location chamber dimensions or mixing assembly is possible For additional information please see the Selection Guides in the U B BPC Catalog Dependent on location in bag and fit with hardware Presentation as dry BPC systems Outer Packaging Supplied flat packed Two polyethylene outer layers Description Product code Lot number Expiry date on outer ing and shipping container Sterilization Irradiation 25 to 38 kGy inside outer packaging Shipping Container Durable cardboard carton Documentation Certificate of Analysis provided with each lot for each delivery Electric Resistive Heater Includes 304 stainless steel outer support con
175. oreactor S U B 101 Thermo Scientific Section 2 7 Pall Kleenpak Connector Instructions With one hand hold the barrel of the male connector Using the other hand grasp both white peel away strips as close as possible to the flat side of connector to ensure a good grip and pull them out simultaneously in a smooth continuous motion Ensure that the connector is perpendicular to the peel away strips shown in illustration for Step e The perpendicular orientation must be maintained while the two strips are pulled simultaneously Do not use if only one white peel away strip is removed accidentally instead of two this will affect the sterility of the pathway Step e Folded over peel away stipes pulled Peel away strips and simultaneously in connector are perpendicular for continuous motion this step in direction as illuatrated lt NOTE Do not impart radial force on the connector A radial force can cause the connector to break If a radial force is present due to items attached to the connector then the connector must be properly supported A f Step f Rotate the thumb rest on the male connector one full turn at least 90 degrees in the clockwise direction The rotation serves to bend the anti actuation tabs which prevents inadvertent engagement If required the thumb rest can be turned counterclockwise to remove kinks in the tubing Minimum 90 degree turn Can be rotated back to remove kin
176. osition Part of Thermo Fisher Scientific ho S OU S os en Ge METTLER TOLEDO 9560608 summing Block il ra Oo ert bls Z h of we n HOLAN a T EL A Se E g 5 ait LOS ASE i a LEARAMCE temae er m AA TOP PLATE p Jaaa CLEARANCE Lr SE CENTER EES E UT THANN Py F AE east F a i BASE PLATE SECTION A 1 Mettler Toledo Panel Mount Display Mettler Toledo Cart or Wall Unlocking Load Cell standard with 1000 LS U B only mount option for 50 500 L S U B and 50 2000 L U M Radial Mounting Load Cells are typically radial mounted Locations for mounting are altered on some units to allow easy access to bottom drain or sparge mechanisms and tubing For more information on load cells to the following documents on the Mettler calibration methods Mettler Toledo terms Toledo Web site or service please go to the Mettler Toledo Web site http us mt com or your local F15175600A Manual for Load Cells representative 16528600A Manual for Panther Display For information on manuals please refer Ordering Information for Single Use Bioreactor and Single Use Mixer Load Cells Retro Fit Kit To Be Load Cells To Be Assembled Volume Assembled By Customer By Manufacturer Description Of Panel Communication Platform 50 amp 100 L SV50194 01 SV50194 11 200 amp 250 L SV50193 01 SV50193 11 500 amp 1000 L SV50218 01 SV50218 11 2000 L SV50227 01 S
177. ow Rate Ipm Versus Vapor Exit Velocity m s 0 61 m s Velocity Threshold C 2 gt p gt a d Flow Ipm e 0 63 0 735 1 00 Threshold Graph 2 2 Thermo Scientific Single Use Bioreactor S U B 81 Section 2 4 single Use Bioreactor BPC Loading 2000 L Water Jacket Various vent filter configurations are available on the S U B depending upon the process scale and intended application Graph 2 2 provides a reference for determining the relative capacity of different filters depending upon the amount of gas flow anticipated and the length of the run In all cases using a vent filter heater will reduce the chance of condensate blocking the filter but over time suspended solids carried in the exhaust stream will impede the flow of exhaust gas resulting in increased back pressure In addition it is good practice to monitor the amount of foam present in the head space In all cases a vent filter heater have very little tolerance for handling the presence of foam in the exhaust stream A small feed of antifoam e g 10 simethicone SH30897 02 added directly to the liquid surface of the culture head space typically provides excellent foam control Large systems especially the 1000 and 2000 L S U B can benefit from the use of a disposable condenser system It has been shown to increase system reliability at high flow rates beyond 50 lpm and should warra
178. ow insert the probe fully as described previously A 5 Position the probe clip in the desired horizontal location Figure 2 145 The clip s adjustable ball detent spring loaded screw should be adjusted so the probe clip can be easily moved to the desired position while still securing the clip to the steel support piece Figure 2 144 Figure 2 145 6 Swing the probe upward into the flexible spring clip Figure 2 146 Release the probe assembly and verify the probe remains at the proper insertion depth and angle as the bellows expands to rest freely in the probe clip Figure 2 147 Figure 2 146 Figure 2 147 NOTE The fixed position of the probe clip and the flexible nature of the spring retainer allow for both proper angle gt 10 degrees and compression of the probe bellows A Single Use Bioreactor S U B 94 Section 2 5 Probe Assembly 2 5 Probe Assembly jd Preparation and Select the appropriate probe see Section 1 Verify the presence of Sterilization a Teflon support ring and O ring on the probe and visually inspect them for damage 2 Perform any required probe maintenance and calibrate the pH probe see Probe Calibration within this section 3 Insert the probe into the probe assembly through the threaded adaptor 4 Verify the probe tip is not touching gt 1 4 6 35 mm gap the membrane of the Kleenpak Connector before threading into probe adaptor 5 Hand tighten the adaptor and v
179. parging systems are now included to provide a wider range of operating conditions This document covers S U B systems with a maximum working volume of 2000 L and the new optimized standard S U B s with dual 3 Sparge capability which provides for a wide range of ka values plus better pH control and lower foaming than the original disk sparger sampling The S U B BPC is supplied Sterile and ready to use Drive Shaft inserted into the S U B BPC through the mixing drive motor and locked into the disposable agitator assembly 4 Process Controls agitation control is 1 Outer Support Container with a mixer drive complete with control unit and a water jacketed tank 2 Single Use Bioreactor BPC complete with agitator assembly spargers overlay and exhaust filter ports plus ports for integration of sensor probes and line sets for media and supplement addition and height diameter ratio Impeller Diameter pH amp DO Probe Autoclavable Type Applisens Broadley James Metler Toledo Hold Up Volume Electrical Power Supply Requirement voltage phase amp Overall Width Overall Length Overall Height integrated into the U B but additional temperature pH and dissolved oxygen DO controls must be supplied 5 Condenser System auxiliary system providing protection against vent filter blockage Guidance data regarding when to use as well as further information are found in the Condenser System data
180. pport shelf e Press downward and into the slots closest to the S U B BPC then slide the holder backward slightly to lock the probe holder into the probe support shelf Probe cap side Figure 2 152 Figure 2 153 NOTE Probe ports that are not used prior to media fill must remain clamped Probe ports not used at the initiation of the run can be accessed later for probe replacement or redundancy purposes if required A Probe Calibration Probe calibration is controller specific however the following general rules apply a pH probes must be calibrated prior to steam sterilization and the calibration of the probe can be standardized by comparison of an off line sample once the pH probe has been connected to the S U B b Dissolved oxygen probes are generally calibrated post steam sterilization Once the probe is connected to the S U B and is allowed sufficient time to polarize 6 8 hours of continuous connection to the power supply provided by a controller or polarization module it can be calibrated Thermo Scientific Single Use Bioreactor S U B 97 2 6 Single Use Bioreactor BPC Sampling Section 2 6 single Use Bioreactor BPC Sampling Sampling During operation of the S U B samples may need to be taken for monitoring of various parameters established by the user The following describes two techniques for sampling aseptic sampling via sterile syringe and sampling utilizing a sterile manifold Aseptic
181. probe RTD is not loose and has been fully inserted into the S U B BPC thermo well Verify thermo well has been filled with sufficient glycerol to aid in heat transfer Verify that temperature controller is enabled Indicator light green LED labeled C1 will illuminate when temperature controller is cycling heater to maintain target temperature Verify mixer is operating safety interlock will not allow heaters to warm without the mixer in operation Verify system is filled with sufficient volume of fluid There must be enough volume of media minimum volume in S U B BPC to provide contact with the bag add more media if BPC is not touching heater area Single Use Bioreactor S U B 186 Thermo Scientific Symptom Remedy Symptom Remedy Symptom Remedy Section 4 Troubleshooting Mixer controller does not respond to user inputs Allow speed to stabilize before using key pad interface Rapidly adjusting speed control in an excessive manner may require several seconds for speed stabilization Wait ten seconds and then attempt to adjust speed at key pad interface Users should verify the position of the input select switch of the VFD If the toggle switch is not in the middle position the controller will not be able to receive control inputs from the control keypad on the front panel Dissolved Oxygen DO readings are low or slow to respond Check physical condition of the DO probe calibration of the DO probe
182. ps Pees eT Table 1 6 Operating Conditions for Cell Culture Applications Full and half volume Gas Supply Setup The following graph depicts a dissolved oxygen management strategy Refer to Table 1 6 for actual gas flow rate recommended maximum values Dissolved Oxygen Control Strategy Air open pipe Air porous frit 0 porous frit 40 60 O Demand Graph 1 1 Dissolved Oxygen Control Strategy Thermo Scientific Single Use Bioreactor S U B 32 Thermo Scientific Section 1 9 5 U B Operating Instructions Open Pipe Sparge Line Frit Sparge Line e Air e Air o 0 e CO e N Figure 1 19 Recommended Gas Inflow Setup In developing the gassing strategy for the dual sparge S U B bag it is recommended that a cascade system be used to maintain dO setpoint starting with the open pipe only air ramping from 0 to 100 output then cascading to air to the frit ramping from 0 to 100 output when needed then finally O to the frit This strategy has allowed excellent CO stripping and high O delivery for high cell densities with little foam formation The cell culture medium used may vary these results refer to Graph 1 1 and Figure 1 19 Single Use Bioreactor S U B 33 Media Fill Agitation Temperature Control Electric Resistive Heating Systems Thermo Scientific Section 1 9 5 U B Operating Instructions For more information on growth of specific
183. r ET pall ACD Connector greater probe membrane longevity os Elevated Filter x e Will restrain probe bellows from ia Pracket a collapsing during sterilization a e Probe holder accommodates two probes Part Number O Auxiliary Components Supporting The Single Use Bioreactor Description Purpose Ouantity Hardware Features 50 L with Resistive Heater The stainless steel outer support container contains the following features 1 Mixer motor 6 Drive shaft 12 Probe shelf 2 Exhaust vent filter holder 7 Liquid sight window 13 Platform with swivel casters and 3 Stainless steel 304 outer support 8 Standard tool set leveling pads container 9 Probe access window 14 Resistive heater w heat shield 4 Bearing port receiver with clamp 10 Control panel 15 Bottom cut outs for BPC alignment 5 Mixing assembly with shield 11 Probe holder 16 Stretch hooks 2 4 a J 3 EN E A T FN 15 ats 16 Hardware Features 50 L with Water Jacket The stainless steel outer support container contains the following features 1 Mixer motor 7 Liquid sight window 13 Platform with swivel casters and 2 Exhaust vent filter holder 8 Standard tool set leveling pads 3 Stainless steel 304 outer support 9 Probe access window 14 Laser dimple water jacket container 10 Control panel 15 Bottom cut outs for BPC alignment 4 Bearing port receiver with clamp 11 Probe holder 16 Stretch hooks 0 Mixing assembly with shield 12
184. r to Section 2 2 a Thermo Scientific Single Use Bioreactor S U B 203 Appendix C Mettler Toledo Load Cell Calibration Instructions Appendix C Mettler Toledo Load Cell Calibration Instructions Load Cell Calibration NOTE Please refer to the instructions and reference material found in the Instructions for S U B Panther Terminal Technical Manual for specific procedures and trouble shooting methods Hardware Systems using Mettler Toledo Prior to reading these instructions confirm the following Panther Display e The Panther Display Load Cell summing block and Load Cell transducers have been specified installed and configured properly e The load cells transducer do not have the transport lock out nuts in place the load cells must be ready for use prior to calibration NOTE The calibration accuracy achieved cannot exceed the precision of the reference used for calibration e Field calibration is most often performed using calibrated reference weights or flow meter for volumetric mass reference e Factory trained technicians have the experience and tools necessary to provide the best system performance and reliability e IFIN DOUBT CONTACT THE FACTORY SERVICE REPRESENTATIVE Introduction e Setup mode is accessed by pressing the select and print keys at the same time these are the two outermost keys See section 3 of the technical manual for further detail e Pressing print is equivalent to ENTER Use th
185. raditional sensors probes that can be integrated into the S U B These requirements are based on the necessary insertion depth of the probe when used with the probe ports Note The presence of a properly positioned O ring on the probe is critical for use with the S U B Recommended pH DO Probes for use with the S U B O ring to probe tip Print Lit Actual Diameter Thread Length Length mm Type mm mm 3 13 5 PG 13 5 PG Probe Lengths from O ring to tip Must Not Exceed 235 mm Probe Part Number Mettler Toledo pH D 2 2 2 19 cO 2 Broadley James DO 2 Broadley James pH Finesse DO Finesse pH 2 22 NOTE Consult probe manufacturer s Web site for appropriate probe cable connection and part number A Table 1 1 Manufacturers and models of U B compatible pH DO probes D de 2 22 239 239 215 133 215 225 225 25 ajs Controllers Many bioreactor controllers can be adapted for use with the S U B Table 1 2 lists several companies offering control systems or custom solutions that will work well when controlling S U B parameters such as dissolved oxygen DO and pH Alternatively users seeking advanced control functionality and data logging capabilities for parameters of pH DO temperature agitation and others can determine their own preferred approach in order to interface these parameters with a single controller Please refer to the Equipment Turnover Package ET
186. rained personnel only Consult manual Electrical components are designed into and required for the proper function of the Single Use Bioreactor S U B The mixer motor motor controller resistive heater jacket and control panel all have electrical components A e Risk of electrical shock and injury e There is potential for static charge build up during mixing and set up of BPCs Warning Entanglement hazard Rotating parts can cause injury Keep hands away from moving parts A Warning Hot surface Do not touch The heating jacket is designed to heat the outer vessel wall Normal operating conditions generate heat and could create hot surfaces A e Hot surface inside e Contact with surfaces may cause burns e Do not touch while in operation Warning Burst Hazard Under normal operating conditions the S U B BPC chamber is under slight pressure Normal passive venting prevents any excess of pressure building up within the chamber Chamber pressure and inlet line pressure should be monitored for proper settings A e Contents under pressure e Do not exceed 0 5 psi 0 03 bar BPC pressure e Do not exceed 5 psi 0 34 bar inlet pressure e Assure vent filter is properly positioned and working properly Thermo Scientific Single Use Bioreactor S U B 1 Protective Earthing Environmental Conditions Water Jacket Vessel Information Thermo Scientific Warnings and Safety Protective earthing must be verified prior
187. robe Kits S U B Temperature Port used to verify RTD calibration Sterile Sampling Manifold with Luer Locks 10 pack 1000 L with Electric Resistive Heater The stainless steel outer support container contains the following features 1 Mixer motor 8 Standard tool set 2 Exhaust vent filter holder g Drive shaft storage holder d Stainless steel outer support 10 Probe access window container 11 Control panel 4 Bearing port receiver with clamp 12 Probe holder 9 Mixing assembly with shield 13 Probe shelf 6 Drive shaft 14 Platform with swivel casters and l Doors 2 leveling pads Top View 15 16 17 18 19 21 Resistive heater with shield 5 Bottom cut outs for BPC alignment Sparge access window Stretch hooks Load cell calibration mount Load cells 3 Load cell summing block 1000 L with Water Jacket The stainless steel outer support container contains the following features Mixer motor 8 Drive shaft storage holder 2 Exhaust vent filter holder g Probe access window 3 Stainless steel outer support 10 Control panel container 11 Probe holder 4 Bearing port receiver with clamp 12 Probe shelf 9 Mixing assembly with shield 13 Platform with swivel casters and 6 Drive shaft not shown leveling pads 7 standard tool set 14 Water jacket Top View 15 16 17 18 19 20 21 Bottom cut outs for BPC alignment Sparge access window Stretch hooks Load cell calibration
188. robe calibration post autoclave can be verified by pulling a sample and analyzing the pH on another calibrated pH meter DO Probe Calibration After polarizing for 6 to 8 hours the DO probe can be calibrated in the S U B using standard protocols In general turn on full air sparging do not exceed 1 lpm in 50 L 1 Ipm in 100 L 2 lpm in 250 L lt 4 lpm in 500 L lt 6 lpm in 1000 L or lt 10 lpm in the 2000 L until the DO reading stabilizes this is 100 DO setpoint for the controller A zero percent dissolved oxygen setpoint can be achieved by sparging with nitrogen or by unplugging the DO probe momentarily Checkpoints Prior to Verify the following before inoculation inoculation V Once S U B is filled with media disconnect BPC tabs from the S U B hardware Verify temperature and agitation are at the desired setpoints pH probe is calibrated autoclaved and connected via a Kleenpak Connector port DO probe is autoclaved connected via a Kleenpak Connector port polarized 6 to 8 hours and calibrated Temperature RTD is completely seated in the thermo well and secured A method for making aseptic tubing connections should be available Ee aa a Sa Thermo Scientific Single Use Bioreactor S U B 35 Cell Inoculation Checkpoints In Process Dispense and Harvest Thermo Scientific Section 1 9 5 U B Operating Instructions Once the S U B is operating at the targeted steady equilibrated state and has achieved te
189. rol unit and an electrical heater The intended use of the Single Use Bioreactor is as of single use bioprocessing without having to buy a or water jacket an animal cell culture bioreactor The HyClone S U B is complete new bioreactor system The critical design 2 Single Use Bioreactor BPC complete with agitator not suitable for use as a microbial fermenter parameters of the S U B such as height to diameter assembly sparger vent filter inlet and outlet ports This document covers the 50 100 250 500 and ratios mixer design and location and typical control plus ports for integration of sensor probes and line 1000 L original standard S U B BPCs with disk sparge system interfaces have been maintained sets The S U B BPC is supplied sterile and ready system In addition to the change to the dual sparge The S U B BioProcess Container BPC is to use system the new standard U B BPC line sets have supplied sterilized by irradiation and therefore does 3 Drive Shaft inserted into the S U B BPC through been optimized to improve functionality Details of not require any facility hook ups for sterilization or the mixing drive motor and locked into the the new standard S U B BPCs along with full details cleaning A key element to the single use design is disposable agitator assembly of the other components of U B systems can be the plastic polyethylene impeller with a bearing seal 4 Process Controls temperature not on water found in dat
190. s Figure 2 105 2 106 Verify that the sparge filter and sparges remain in position while attaching the tabs It is recommended that users secure the tabs on the front BPC panel first This way the larger rear door will allow access when connecting the last set of tab pins Thermo Scientific Single Use Bioreactor S U B 76 Section 2 4 single Use Bioreactor BPC Loading 2000 L Water Jacket Figure 2 105 Figure 2 106 16 While the BPC begins to fill with air remove the protective packaging from the exhaust vent filters If the bag is filled sufficiently with air the filters can be loaded into the holder e If using the exhaust condenser system follow the setup instructions in Section 2 4 e If using elevated exhaust vent filters use the corresponding extended dual vent filter bracket and filter heaters Follow the setup instructions in Section 2 4 Thermo Scientific Single Use Bioreactor S U B 71 Section 2 4 single Use Bioreactor BPC Loading 2000 L Water Jacket Disposable Exhaust Condenser Condenser System The condenser system is intended to be used as an accessory to the Functional Overview Single Use Bioreactor S U B as an alternative to vent filter heaters The condenser s purpose is to prevent liquids and solids from condensing and collecting inside of the vent filters of the S U B The condenser system cools the exhaust gasses leaving the S U B chamber condensing the moisture out of the saturated gasses comi
191. s for cell culture vary greatly between cell lines and media formulations Table 1 5 is provided as a reference for establishing initial control settings for the S U B Proper management of gas flow rates is a critical parameter when performing cell culture in a BPC It is therefore recommended the end user carefully consider the gas flow rate introduced into the S U B The gas flow rates listed here are recommended maximum rates If the oxygen demand of the culture exceeds that delivered by these rates the gas ratios can be adjusted accordingly to deliver more oxygen e It is strongly recommended not to exceed the total maximum gas flow rate into the bag Otherwise these high sparge flow rates will result in excessive foaming and increased load on exhaust vent filters e A sparge strategy that uses an initial flow rate of air and then cascades supplemental oxygen has been shown to be a highly effective approach in maintaining the desired oxygen saturation setpoint This is typically the most efficient strategy for meeting the respiration demands of high density cell culture while reducing the risk of creating excessive pressure in the S U B Lower flow rates provide two important benefits 1 reduced generation of foam in the headspace area and 2 reduced pressure loading on the S U B BPC and exhaust vent filter When using oxygen cascade flow rates can typically be reduced four to five times as compared to air because air typically cont
192. sher Scientific Inc All rights reserved SmartSite is a trademark of ALARIS Medical Systems Inc C Flex and PharMed are registered trademarks of Saint Gobain PharmaPure is a registered trademark of PharmaPure Tubing Clave is a registered trademark of ICU Medical Pall Kleenpak and HyClone Laboratories Inc Logan UT USA is ISO Certified Emflon are registered trademarks of Pall Corporation Lynx is a registered trademark of Millipore SterilEnz is a registered trademark of PAW Bioscience Products INC All other trademarks are the property of Thermo Fisher Scientific Inc and its subsidiaries Specifications terms and pricing are subject to change Not all products are available in all countries Please consult your local sales representative for details Cell Culture amp BioProcessing 925 West 1800 South In Americas Asia In Europe www thermoscientific com hyclone Logan UT 435 792 8000 32 53 85 71 80 84321 435 792 8001 fax 32 53 85 74 31 fax SCIENTIFIC Section 3 8 Condenser System Data Sheet 3 8 Condenser 3 8 1 Condenser System Data Sheet System Data Sheet Thermo Scientific Single Use Bioreactor S U B 174 The Thermo Scientific HyClone Single Use Bioreactor S U B Condenser System specifically supports the effective use of the HyClone 2000 L S U B and is also available as an auxiliary product for all other S U B systems It efficiently condenses exhaust gases and transfers condensate back into the
193. still in polybags over the side of the outer support container This will help support the weight of the BPC and also keep the BPC from being restricted during the air inflation step Load the BPC bearing port into the receiver Figure 2 67 and close the door and clamp Figure 2 68 Figure 2 67 Figure 2 68 Thermo Scientific Single Use Bioreactor S U B 63 Section 2 3 single Use Bioreactor BPC Loading 1000 L Water Jacket 6 Connect pressure transducer to monitor Once the display has stabilized tare the monitor the monitor should be allowed to warm up for 30 minutes before taring Verify that the monitor reads zero Gas Supply The BPC should be partially inflated in order to aid in proper alignment Connections and of the BPC in the outer support container and will be necessary to allow for proper insertion of the drive shaft Air Inflation 7 Attach air supply to overlay gas inlet line at the top of the BPC NOTE Air pressure to the overlay gas line on the S U B BPC should not exceed 3 psi less than 25 lpm 4 WARNING DO NOT EXCEED 0 5 psi 0 34 bar within the BPC or the system could fail causing personal injury or damage to equipment DO NOT leave the BPC unattended while inflating a NOTE Air pressure to the sparger on the S U B BPC should not exceed 8 psi Also while a sparge line check valve is provided for each sparge line it is not uncommon for some fluid to bypass check valves during typical use
194. t or swapping overlay inlet line with supplement line Dual sparger open pipe plus porous frit standard Can do an engineer to order for one or the other alone Make to order bags will be built around standard dual sparger chamber Limited customization possible Suggested use through 1 port so Length cannot interfere with impeller and shaft this is engineer to order Otherwise must use ferrule approach Typical is 10 or shorter Can use disposable capsule filter other than standard hydrophobic vent filter with Emflon Il standard is Pall Kleenpak 0 2 micron exhaust vent filter Note Vent filter heater configuration restricts options Extended filter height above the U B bag is make to order Must be compatible with a vent filter bracket option Choice of filters for inlets used to sterile filter incoming media or supplements Please Note Not all options are available for all ports No customization of port type and location chamber dimensions or mixing assembly is possible For additional information please see the Selection Guides in the U B BPC Catalog Dependent on location in bag and fit with hardware Presentation as dry BPC systems Description Product code Lot number Expiry date on outer and shipping container Irradiation 25 to 38 kGy inside outer packaging Durable cardboard carton Certificate of Analysis provided with each lot for each deliver SV 50197 01 100 L S U B US version 120 VAC Singl
195. t sampling or attachment of various sampling manifolds by use of standard luer lock connection Alternatively manifolds can be welded onto the C Flex sample line by tubing welder BPC drawings and standard configuration tables specific to volume can be found in Section 3 Single Use Bioreactor S U B 10 Section 1 4 BioProcess Container Characteristics BPC Features SQ Figure 1 2 Typical 50 L 100 L Figure 1 3 Typical 500 L and 1000 L BPC Figure 1 4 Typical 2000 L BPC and 250 L BPC Schematic schematic schematic Part Description Exhaust Vent Filter single use capsule filter for exhaust gas exchange Gas Overlay Port Protected by gas filter For addition of media and other liquids 1 A E E Links with mixer motor and allows impeller to turn while J y retaining integrity of the S U B BPC facie Injection molded plastic Linked to seal bearing assembly by P C Flex tubing contact material of the shaft Ports with Kleenpak For integration of standard 12 mm monitoring pH and DO Connectors probes For integration of temperature probe while retaining e f Operatu E ean orie SUB BPC Sampling Port For needleless sampling or connection to sampling manifold Used when draining the S U B 0 Gas Sparge Lines Sparge integrated into the chamber Protected by gas filters Integrates with chill plate to remove condensate from exhaust Table 1 3 Key to Figures 1 2 1 3 and 1 4 Condenser System Bag o
196. t tension on the latch is less than seen when the latch was first closed and adjust the threaded latch pin sufficient to allow the latch handle to begin to tension Release latch 3 and adjust the tension as in the previous step Release latch 1 and adjust the tension and close Release latch 4 and adjust the tension and close For maximum security insert safety pins not included into the respective latches Once the drive shaft is inserted and side access doors closed turn off the air supply to the overlay line If the optional exhaust vent filter heaters are being used secure heaters to the filter at this time Single Use Bioreactor S U B 60 Section 2 3 single Use Bioreactor BPC Loading 1000 L Electric Resistive Heater Final Installation Ste ps Secure exhaust vent filters on top mounted holders by use of the filter side vents Figure 2 61A or if using elevated dual exhaust filters use the adapter piece and extended filter bracket Figure 2 61B Figure 2 61B Extended Dual Filter Bracket 37 Fully extend the drain line set through the front cutout and attach the probe shelf 38 Remove the poly bag from the drain line set and position the line clamp as close as possible to the BPC port and close Use a cable tie around the clamp to ensure the clamp cannot be accidentally opened 39 Align the five Kleenpak ports through the front access window Figure 2 62 _ Figure 2 61A Figure 2 62 Thermo Scientifi
197. tainer with swivel caster anaye COO U B URNES IONAN AVAG Ingle RARR platform variable speed agitation controller motor drive assembly with shaft PID temperature controller RID sensor integrated resistive heating 8V50172 02 250 L S U B EU version 240 VAC Single Phase element probe shelf and standard tool set Water Jacket Includes 304 stainless steel outer support container with swivel caster platform variable speed agitation controller motor drive assembly with 9V50172 03 250 L S U B US version 120 VAC Single Phase SV50172 04 250 L S U B EU version 240 VAC Single Phase ou RTD sensor integrated water jacket probe shelf and standard tool Custom Hardware Options For stainless steel capabilities please contact your sales representative for HyClone products for more information ee Used to package the pH and DO probes not supplied for sterilization SMAAD Piaroa oton POOR AS SEMON NOn SEE and to aseptically connect them to the S U B BPC See Figure 1 SV20664 01 Heavy Duty Tubing Clamp ad l er f SV20664 02 Heavy Duty Tubing Clamp 4 pack of SV20664 01 1949891 tubing camp used on Probe poris when connecting Prove assemblies SV20664 03 Heavy Duty Tubing Clamp 10 pack of SV20664 01 SV20750 01 S U B Temperature Sample Port Used for RTD calibration validation SV50177 01 Autoclave Tray for Probe Kits stainless steel sae Ue i AGNES TICE DIDR eee Desi autoclave 0ee 5H30845 01 Sterile Samp
198. tems Section 1 8 5 U B Operation Information The agitator should not be operated at volumes less than the stated minimum volume Stirring speed is adjusted by using the keypad interface on the control panel The agitation control interface utilizes a red LED digital display to indicate stirring speed in units of revolution per minute RPM Power is supplied to the motor by a two position power switch that is illuminated in green when turned to the ON position right position The up and down arrows on the agitation keypad adjust the stirring speed Due to the auto restart capabilities of the S U B the green start button on the keypad has been disabled however the red stop button on the keypad is active If the red stop button has been used to stop the motor the controller can be reset and agitation restarted by using the main motor toggle switch on the left side of the control panel The S U B is equipped with a bottom drain line that allows for liquid harvest by means of peristaltic pump Connection of the bottom drain line can be accomplished by use of a tubing welder or the aseptic connection of 1 4 3 8 6 35 9 53 mm quick connect that is provided For S U B s 50 L to 1000 L the bottom drain exits the U B BPC at the lowest vertical position on the side of the S U B This allows for easy access for the user and minimizes the accumulation of cells in the area of the drain during the cell culture run Manipulation of the
199. tents Troubleshooting eooooeeeeeeeeeesseeeeeerrrnnssessssssssssssseeeerrrreeesssessss 186 Ordering Information 5 1 Ordering Information Disposables BPC seeeeseeeeereseseseresssress 193 5 2 Ordering Information Hardware nessssseicsreseisviciesesiiesi 194 5 3 Ordering Information Accessories ssssssesesesessseressressressresssrsses 197 5 4 Ordering Information Partial Spare Parts List eeeeeeeeeeeeees 198 39 Ordering nist ri ctionis sxass Ge essesatepvedetectacesatsieagOeaesienadeusee eames 200 5 6 Ordering Support Contact Information eeeeeeseeseeeeeneeeeneeeees 200 Electrical Receptacle Installation Instructions eeeeeeeseeeeeeneeeeeees 201 Calibration of the AC Tech variable speed drive ceeeeeeesseeeesteeeees 203 Mettler Toledo Load Cell Calibration Instructions ccccescceeeeee 204 Single Use Bioreactor S U B iV Warnings and Safety Warning Read and understand operator s manual before using this equipment Thermo Scientific HyClone Single Use Bioreactor is designed to be operated under traditional mammalian cell culture conditions A general before using this understanding of bioreactor systems and their operation is important prior machin to using the system for the first time A e Read and understand users manual before operating e Failure to do so could result in injury Warning Hazardous voltage inside Disconnect power before opening Service by t
200. ter ratios mixer design and location and typical control system of ka values better pH control and lower ready to use interfaces have been maintained foaming than the original disk sparger 3 Drive Shaft inserted into the U B BPC The S U B BioProcess Container BPC is supplied sterilized through the mixing drive motor and locked into the disposable agitator assembly by irradiation and therefore does not require any facility hook ups for 4 Process Controls temperature not on sterilization or cleaning A key element to the single use design Is the plastic polyethylene impeller with a bearing seal assembly linked to 1 Outer Support Container with a mixer water jacket and agitation control are arotenalmkerdie drive complete with control unit and an integrated into the S U B but additional electrical heater or water jacket pH or dissolved oxygen DO controls must 2 Single Use Bioreactor BPC complete be supplied by the user Rated Liquid Working Volume Minimum Liquid Working Volume Total Reactor Volume Liquid amp Gas BPC Chamber Diameter BPC Chamber Shoulder Height Liquid Height Rated Working Volume Overall Reactor Geometry height diameter ratio Fluid Geometry Working Volume 151 height diameter ratio mpeller quantity x blade count Impeller Diameter 7 88 20 cm 7 88 20 cm pH amp DO Probe Autoclavable Type 12 mm diameter x 215 235 mm Applisens Broadley James Metler Toledo insertion
201. the outside edge of the latch base Figure 2 86 NOTE When the latch is under tensioned the safety pin pass through of the latch handle will be covered within the latch base and the handle will close very easily If the latch is over tensioned the handle will be excessively difficult to close Outside Edge of Latch Base safety Pin Pass Through of Latch set Latch Retainer safety Pin Pass Through of Latch Handle Figure 2 86 Thermo Scientific Single Use Bioreactor S U B 70 Section 2 3 single Use Bioreactor BPC Loading 1000 L Water Jacket 27 For maximum security insert safety pins not included into the respective latches 28 Once the drive shaft is inserted and side access doors closed turn off the air supply to the overlay line If the optional exhaust vent filter heaters are being used secure heaters to the filter at this time Final Installation Ste ps 29 Verify position of the exhaust vent filters on top mounted holders by use of the velcro fasteners Figure 2 87 Figure 2 87 30 Fully extend the drain line set through the front cutout and attach the probe shelf 31 Remove the poly bag from the drain line set and position the line clamp as close as possible to the BPC port and close Use a cable tie around the clamp to ensure the clamp cannot be accidentally opened 32 Align the five Kleenpak ports through the front access window Figure 2 88 This will be the lower cutout if hardware is supplied
202. tial to provide continuous and nearly indefinite protection against performance degradation of the exhaust vent filters rated for continuous flow rates up to a total off gassing rate of 50 lpm and intermittent flows up to 100 lpm See Single Use Bioreactor System Validation Guide Thermo Scientific Single Use Bioreactor S U B 79 Section 2 4 single Use Bioreactor BPC Loading 2000 L Water Jacket Table 2 1 can help end users specify the BPC configuration and operating parameter setup for custom 2000 L S U B applications when not utilizing the disposable exhaust condenser Because the operating parameters of different cell cultures vary widely a safety factor should be used to temper the data As such the data we used to generate a control base line are for reference only filter fouling will vary and must be considered to ensure reliable performance It is assumed no foam is present in the exhaust stream S U B System Max Combined Flow Resulting Safety Run Duration 2 ea 10 vents Rate Recommended Factor 2000 L S U B 40 Ipm 2X 2000 L S U B 14 days 27 Ipm 3X 2000 L U B 21 days strongly Recommend Single Use Condenser Table 2 1 Condenser System Overview 2000 L S U B 10 days 32 Ipm 2 5X The above recommendations were generated using the test conditions shown in Graph 2 1 In this case a 2000 L S U B was filled with 2000 L of DI water with a batch temperature of 40 C using a MKS vent filter heater at 60 C Safe
203. ting parameters yet it has the unique advantage of being a completely disposable device Ease of setup with respect to system operation and integration into existing facilities makes the S U B an attractive alternative to its stainless steel counterpart Critical design parameters such as height to diameter ratios mixer design and location and typical control system interfaces have been maintained The S U B BPC is supplied sterilized by irradiation and therefore does not require any facility hook ups for sterilization or cleaning A key element to the single use design is the plastic polyethylene impeller with a bearing seal assembly linking to an external mixer drive Quick setup and change over allows for faster turnover in cell culture runs over traditional reusable systems The S U B systems consist of three primary components 1 Outer support container with either Electric Resistive or Water Jacket heating systems 2 Single Use Bioreactor BioProcess Container BPC and 3 Control System for agitation Electric Resistive and Water Jacket and temperature resistive only The outer support container is engineered and fabricated to fully support each S U B BPC and allow easy access for operation It is a stainless steel vessel which holds and supports the BPC The outer support container contains the mixing drive silicone heating blanket Electric Resistive or Water Jacket controllers for mixing and heating Electric Resistive
204. ting volume Serious harm to the operator or damage to the S U B BPC will result a General temperature mapping has been performed for the S U B systems by tracking thermal profiles within the liquid confines of the bioreactor Testing conditions were analyzed using chilled media and low agitation rates Gradients within the liquid did not exceed 0 5 C during heat up Testing has also shown that temperature measurements in the S U B when using the standard silicone thermo well with 1 8 3 18 mm diameter temperature probes property represent bioreactor content temperatures Users desiring exact temperature calibrations can order S U B temperature sample port See Section 3 9 Using this port will allow users to simulate the temperatures seen by the RTD when used with the S U B BPC Single Use Bioreactor S U B 28 Protective Earthing Replaceable Fuses Thermo Scientific Section 1 8 5 U B Operation Information Heating Times for S U B systems ambient temperature of 25 C values for jacketed systems assume a TCU heater size of 10W per batch liter Liquid Batch Volume Initial Liquid Time half full Liquid Target half full 5 hrs 7 5 hrs 3 hrs 2 5 hrs 5 hrs 7 5 hrs System Electric 50 L 25 L minimum 50 L Jacketed 50 L 25 L minimum 50 L Electric 100 L 50 L minimum 100 L Jacketed 100 L 50L minimum 100 L Electric 250 L 125 L minimum 250 L Jacketed 250 L Jacketed 500 L Ja
205. tion Single Use Bioreactor S U B 192 Section 5 1 Ordering Information Disposables 5 1 Ordering Information Disposables Standard BPC Products PartNumber Description Additional Information 9H30774 01 50 L S U B BPC 9H30774 02 100 L S U B BPC 5H30774 03 250 L S U B BPC and a porous frit sparge All SE O a gaming INE Custom BPC Products Thermo Scientific Category Options Capability Remarks Tubing Type C Flex platinum cured silicone PVC Pharmed PharmaPure More information in selection guide of BPC catalog Tubing Size Specific lengths of 1 8 ID to 1 ID ID limitations due to port size Luers CPC Quick Connects SIP connectors Triclamp Kleenpak Lynx Refer to selection guide in BPC catalog for further Connectors SmartSite Clave Lynx steam thru CPC Steam Thru Gore steam information by connection type Note Only option valve Gore Mini TC BioQuate GE PAW SterilEnz end plug etc for probe port connections is Kleenpak MET Ea feet Additional ports second row of 4 for 50 L to 250 L S U Bs second ports row of 5 for 500 L and 1000 L U B s Pressure sensor Pendotech for 50 L to 250 L S U B comes standard on 500 L and 1000 L Disposable sensors DO Finesse DO and pH PreSens Availability of various other disposable sensors currently pending Addition of ports lines other than 2nd row of probe ports a Da TEET E ET Dependent on location in bag and f
206. tion step 8 Remove the black protective cap from the bearing port Figure 2 96 load the BPC bearing port into the receiver Figure 2 97 and close the bearing assembly door and clamp Figure 2 98 a i 3 Figure 2 98 9 Open the tubing set polybag and connect pressure transducer to monitor Once the display has stabilized tare the monitor the monitor should be allowed to warm up for 30 minutes and the sensor connected for 10 minutes before taring Verify that the monitor Thermo Scientific Single Use Bioreactor S U B 74 Section 2 4 single Use Bioreactor BPC Loading 2000 L Water Jacket Gas Supply reads zero Connections and Air The BPC must be partially inflated in order to aid in proper alignment Inflation of the BPC in the outer support container and is necessary to allow for proper insertion of the drive shaft 10 Attach air supply to overlay gas inlet line at the top of the BPC Operators should allow the bag to fill to greater than half volume typically gt 20 minutes NOTE Air pressure to the overlay gas line on the S U B BPC should not exceed 5 psi less than 100 lpm A WARNING DO NOT EXCEED 0 5 psi 0 34 bar within the BPC or the system could fail causing personal injury or damage to equipment DO NOT leave the BPC unattended while inflating A 11 Feed the probe belt sample line and the subsurface addition lines through the front access door Figure 2 99 12 Remove sparge lines from the
207. tional this configuration 2 Clip each filter one at a time into the elevated vent filter holder system carefully center the filter housing allowing the clip to secure it near the hose barb connections Figure 2 119 3 Verify the routing of the exhaust tubing ensuring that it is not likely to cause kinks in the tubing 4 Place the vent heaters around each filter ensuring the snap retainers are secured Position the power leads to avoid interfering with the vent holder brackets Figure 2 120 5 Raise and rotate the vent holder bracket as needed Make final inspection to ensure no kinks or low spots will occur in the tubing between the BPC and the filter even if the BPC becomes pressurized Figure 2 121 6 Connect the power to the vent heaters and verify operation of controllers 7 Inspect the controllers set points recommended 60 C After 2 5 minutes of operation verify the vent heaters are warm and near the desired temperature set points and that no alarm indicators are active F Figure 2 119 Figure 2 120 Figure 2 121 Thermo Scientific Single Use Bioreactor S U B 86 Drive Shaft Insertion 1 The drive shaft is constructed by assembling four pieces these must be assembled and inserted piecewise in areas having low ceilings 2 Reminder The BPC must be filled with enough air to allow for unrestricted loading of the angled drive shaft Operators should be elevated i e ladder to effect
208. tive for HyClone products for more information aes Used to package the pH and DO probes not supplied for sterilization prune BIO A OL RODS WEN ets ele and to aseptically connect them to the S U B BPC See Figure 1 SV20664 01 Heavy Duty Tubing Clamp SV20664 02 Heavy Duty Tubing Clamp 4 pack of SV20664 01 ooe clamp used on probe ports when connecting probe SV20664 03 Heavy Duty Tubing Clamp 10 pack of SV20664 01 l SV20750 01 S U B Temperature Sample Port Used for RTD calibration validation SV50177 01 Autoclave Tray for Probe Kits stainless steel n Lry ANNICA AOAN MIC DES NA De alan aller average SH30845 01 Sterile Sampling Manifold with luer lock individual Available for aseptic luer connection or weld onto the sample line to SH30845 02 Sterile Sampling Manifold with luer lock 10 pack take sample sets SV50177 65 Frit Sparge Line Support Recommended to maintain the frit sparge in a vertical position SV50177 16 Elevated Filter Bracket Needed to support an elevated vent filter Ny Probe Bioreactor Probe Autoclave Tray a Frit Sparge Line Assembly for Probe Kits d l PO die Support f a Threaded Probe Attachment W C cable Tie Bioreactor Probe SA h 2 Molded Bellows Autoclave Tray t sE Cover _ Additional information on autoclave tray n e Fabricated from stainless steel be Cable Tie e Plastic carry handle for easy transport S gt right out of the autoclave ach e Positions probes on 15 incline fo
209. ty factor estimates are based on a maximum continuous internal S U B BPC pressure not to exceed 0 1 psi which corresponds to 40 lpm with a single 10 vent 2000 L S U B BPC Pressure Trending Direct Sparge Gas Loading 2000 L DI Water 40 C Vent Heater 60 C Overlay Pressure psi 40 50 Air Sparge Ipm e Single Meissner 10 Vent 0 2 micron PVDF Graph 2 1 NOTE These results do not take into consideration a fouling safety factor A Thermo Scientific Single Use Bioreactor S U B 80 Section 2 4 single Use Bioreactor BPC Loading 2000 L Water Jacket Also consider the size and type of tubing used to connect the exhaust vents to the S U B BPC when not using an exhaust condenser Braid reinforced tubing provides the best protection against kinking or accidental pinching of the exhaust line The 10 vents are supplied with 34 hose barbs and this tubing diameter will allow condensate to return to the S U B at total off gas flow rates up to 30 lpm assuming the tubing is near a vertical orientation Testing has shown that large diameter tubing will allow for lower exhaust gas velocities and if the vapor velocity is below 0 6 m s gravity will allow the condensate formed in the tubing to return to the batch process NOTE Restrictive tubing connectors can create flow bottle necks and Y ID tubing is typically deemed too small for the 2000 LS U B A Tubing I D Recommendation inches Fl
210. ual Available for aseptic luer connection or weld onto the sample line to SH30845 02 Sterile Sampling Manifold with luer lock 10 pack take sample sets 9V50177 65 Frit Sparge Line Support Recommended to maintain the frit sparge in a vertical position SV50177 16 Elevated Filter Bracket Needed to support an elevated vent filter My Probe Bioreactor Probe Autoclave Tray Frit Sparge Line Assembly for Probe Kits p Se Handie Support En Threaded Probe NX Attachment kA C cable Tie Bioreactor Probe S b Autoclave Tray Additional information on autoclave tray e Fabricated from stainless steel e Plastic carry handle for easy transport 7 right out of the autoclave A e Positions probes on 15 incline for greater probe membrane longevity p e Will restrain probe bellows from t collapsing during sterilization e Probe holder accommodates two probes Elevated Filter s Bracket i Pall ACD Connector SV50197 100 L SH30774 02 SH30720 01 SV20664 03 SV50177 01 S U B Hardware System standard S U B BPC standard Bioreactor Probe Assembly non sterile for use in autoclave Heavy Duty Tubing Clamp 10 pack Autoclave Tray for Autoclaving Probe Kits SV20750 01 SH30845 02 S U B Temperature Port used to verify RTD calibration Sterile Sampling Manifold with Luer Locks 10 pack Necessary for regulation of gas pressure and flow rate control for DO and pH set points Autoclavable Probe
211. uer Locks 10 pack Necessary for regulation of gas pressure and flow rate control for DO and pH set points Autoclavable Probe 13 mm X 13 5PG thread w 195 235 mm insertion length Autoclavable Probe 13 mm X 13 5PG thread w 195 235 mm insertion length Tubing Welder Terumo or Wave Steam In Place SIP Sterilizer or Laminar Flow Hood Used for fluid transfer between line sets and containers Necessary for water jacket temperature control 2000 L with Water Jacket The stainless steel outer support container contains the following features Mixer motor Mixing assembly with shield External E Stop assembly Stainless steel outer support container D Bag lift assembly a Le Standard tool set Load cell display Control panel Load cell summing block Load cell 3 Probe access window 6 7 8 s1 1 1 ODO 12 13 14 15 16 17 Probe clips Water jacket Bottom cut outs for BPC alignment Sparge access plate Jacket quick connect couplings Pneumatic bag lift control Condenser System The Condenser System contains the following features Chiller Plate Pump assembly Electrical power strip Filter brackets Condenser cart assembly Condenser post filter bracket assembly Chiller temperature control unit TCU Chiller line assembly PO N S o N o gi ae 75 12 71 55 1817 3 1908 2 47 05 41 05 1195 0 40 17 10
212. ulture amp BioProcessing 925 West 1800 South In Americas Asia In Europe www thermoscientific com hyclone Logan UT 435 792 8000 32 53 85 71 80 84321 435 792 8001 fax 32 53 85 74 31 fax ha Gan a a a a a O 3 6 2 2000 L Hardware Specifications Thermo Scientific Water Jacket Section 3 6 2 2000 L Hardware Specifications Water Jacket fakes E npr Scainginpelerdanetefinkdaneted IB Space Quick Connect Check Valves Ambient to 40 0 1 C 104 0 2 F Asyawoay 10373983Y ETEL siajaweleg U018 Y joujuo0s dwaj sauiejuoy Woddns TENER Single Use Bioreactor S U B 166 Section 3 6 3 2000 L Control Panel Layout 3 6 3 2000 L Control Panel Layout Front View Side View Power On Button Light a oOo ha i Main Power Source 16 pin Communication Quick Connect speed Control Key Pad Motor Controller On Off Switch External E Stop Connection Emergency Stop RTD Input Signal M i h ji i Ta Il Motor Cable Bottom View Buzzer Spare 14 Liquid Tight RTD Input Signal Emergency Shut off Power This outlet is used for devices that should be turned off when an operator activates an E stop Examples are pumps motors or any device that could potentially harm an operator or ruin a batch during an E stop The max for each receptacle is 2 amps Continuous Power This outlet is used for devices that should not be turne
213. use condenser system This allows the S U B to utilize a powerful phase change type system which provides improved exhaust vent protection and reliability due to the ability to strip condensate and atomized materials that may be present from the off gas stream of the S U B This system has been shown to significantly reduce the fouling load on the vent filters that inherently increases operating back pressure as the cell culture run batch progresses See Single Use Bioreactor Validation Guide for specific details 2000 L S U B BPC with Some end users may prefer to omit the Condenser System on the 2000 Vent and Heaters Only L S U B under the expectation that this will allow for a more uniform installation similar to smaller S U B s used in the upstream seed train or will perhaps reduce system complexity and cost The use of exhaust vent heaters and 10 filters will provide some impressive flow capacity over short periods lt 5 days However the high sparge rates required during the scale up of the S U B to the 2000 L working volume may eventually create conditions of increased operating back pressure usually due in part to blocking of the filter media Depending upon the application the user has the option of using both filters in parallel or initiating the run with a single filter temporarily clamping off the line to the other filter it being reserved as redundant back up NOTE The disposable Condenser System has the poten
214. volume A Heating times for the Single Use Bioreactor systems vary based upon liquid volume and temperature ambient or heating liquid temperature sparge rate and mixing rate Users should adjust process liquid staging and seeding strategies to the unique aspects of the S U B Process controllers and heaters are designed to provide optimum heat transfer and to minimize heat up times while maintaining the material integrity of the polymer film construction of the S U B BPC see Table 1 5 Do not operate heater if S U B BPC is not at minimum liquid volume or greater Care must be taken not to melt or damage the bag or other disposable components of the S U B BPC Heaters should not be used to warm liquid above 40 C In order to reduce the possibility of the tank becoming too hot the heaters are supplied with a thermostat that will limit temperature to 65 5 C The main power disconnect should be in the off position unless the S U B hardware system is in active use This will preserve the long term reliability of the heating system Note Users can reduce the heating time required to warm media by allowing media to warm to ambient temperatures before transferring into the S U B BPC WARNING The heating element is provided with an over temperature safety that will prevent direct damage of the heater element The heater must not be operated when the outer support container is empty or when the S U B is below minimum liquid opera
215. with two horizontal slots Figure 2 88 Thermo Scientific Single Use Bioreactor S U B 71 Thermo Scientific Section 2 3 single Use Bioreactor BPC Loading 1000 L Water Jacket 33 Insert RTD or selected temperature sensor into the thermo well Figure 2 89 a Place a small amount of glycerol 0 5 mL in well to aid in heat transfer Glycerol Sigma G6279 b Sensor should be inserted until base of RTD meets the mouth of the thermo well c Secure by twisting the luer lock collar if provided The thermo well will stretch slightly when RTD is seated Figure 2 90 Figure 2 89 Figure 2 90 NOTE Verify all port clamps are closed and located as near as possible to the body of the S U B BPC a Single Use Bioreactor S U B 12 2 4 Loading 2000 L Water Jacket Figure 2 91 Thermo Scientific l Section 2 4 Single Use Bioreactor BPC Loading 2000 L Water Jacket Main power to the control panel should be switched on Ensure that the drive motor is not running Open both the front and rear doors on outer support container and use an elevated platform to open clamp on bearing port receiver located below the motor Figure 2 91 Lower the hoist lifting frame to a position just above the top of the rear door by using the pneumatic control lever near the rear door Two operators should carefully remove irradiated S U B BPC from protective double polybags Do not remove the polybags from the line sets at this stag
216. ypical use Elevating the filter to ensure that it is not at the low point of the sparge line will reduce the chance that the filter is exposed to liquid a Lii A Figure 2 11A Membrane Sparge Figure 2 11B Aligning Dual Positioning Sparge in Base Slot Thermo Scientific Single Use Bioreactor S U B 41 Section 2 2 single Use Bioreactor BPC Loading 50 500 L Volumes 10 Use the four bottom cut outs located at the base of the support container as a reference to align hanging tabs on S U B BPC Figure 2 12 Figure 2 12 11 Attach platform stretch hooks to each of the four bottom corners of the BPC Figure 2 13 Figure 2 13 12 Position bottom side drain pulling out downward to position port towards bottom edge of tank 13 Align row of Kleenpak Connector probe ports within access window Figure 2 14 Figure 2 1 NOTE Verify all port clamps are closed and located as CLOSE as possible to the body of the S U B BPC a Thermo Scientific Single Use Bioreactor S U B 42 Thermo Scientific Section 2 2 single Use Bioreactor BPC Loading 50 500 L Volumes NOTE Air fill of the S U B BPC takes approximately 10 20 minutes to allow for drive shaft insertion times will vary based on flow rate inlet pressure and bag volume The BPC must be at least partially inflated in order to insert drive shaft A 14 Remove the latch pin from the safety cover over the mixing assembly and open the cover Unscr

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