ÄKTAdesign Reverse flow Kit - GE Healthcare Life Sciences
Contents
1. Any 0 00 Block Start_Conditions Start_Conditions 0 00 Base SameAsMain 0 00 Wavelength 280 Wavelength_1 nm OFF i Wavelength_2 nm OFF ifWavelength_3 nm 0 00 AveragingTimeUV 2 56 UV_Averaging_Time sec 0 00 Alarm_Pressure Enabled 4 00 Pressure_Limit MPa 0 00 MPa 0 00 OutletValve WasteF1 fH 0 00 Block Initial_Eluent_Conditions 0 00 End_block H 0 00 Block Column_Equilibration 0 00 Block Flowthrough_Fractionation 0 00 Block Sample_Injection 0 00 Block Wash_Out_Unbound_Sample 0 00 Block Eluate_and_Peak_Fractionation 0 00 Block Gradient_Elution 0 00 Block Clean_After_Gradient Click here to define a 0 00 Block Reequilibration 0 00 End method variable name for the instruction parameter Sen so inectcr aive ES Eee Pump E Insert OutletValve Vi mm io oT BuffeMalveA1 va DownFiow C UpFiow Tae ce yi lowpath Pumpalnlet change _ Mes C Alarms amp Mon PumpBInlet e IniectionMark C Watch C Other ao Delete Ready NUN 3 Click the Flowpath radio button 4 From the Instructions list select FlowDirection The valve instruction parameter Position allows you to change position for the flow direction valve using parameters DownFlow and UpFlow 26 Reverse flow Kit User Manual 71 5029 22 Edition AC Operation 7 5 Define a variable name for the instruction parameter e g FlowDirection This variable allows you to turn the position of the valve to the direction you2. 1 nm OFF Wavelength_2 nm OFF Wavelength_3 nm 0 00 AveragingTimeUV 2 56 UY_Averaging Time sec 0 00 Alarm_Pressure Enabled 4 00 Pressure_Limit MPa 0 00 MPa 0 00 OutletValve WasteF1 Mi 0 00 Block Initial_Eluent_Conditions 0 00 End_block 0 00 Block Column_Equilibration Mi 0 00 Block Flowthrough_Fractionation Mi 0 00 Block Sample_Injection W 0 00 Block Wash_Out_Unbound_Sample MM 0 00 Block Eluate_and_Peak_Fractionation W 0 00 Block Gradient_Elution W 0 00 Block Clean_After_Gradient Click here to define a W 0 00 Block Reequilibration j 0 00 End_method variable name for the instruction parameter r Instructions InjectionValve Position Insert CP e Ce ot a vomon C Up Flowpath ufferV aly 7 Pp Pump lnlet TER Mi AlarmstMon PumpBInlet IniectionM ark Frac Replace C Watch C Other a Delete Ready NUM 3 Click the Flowpath radio button 4 From the Instructions list select FlowDirection The valve instruction parameter Position allows you to change position for the flow direction valve using parameters DownFlow and UpFlow Reverse flow Kit User Manual 71 5029 22 Edition AC Operation 7 Define a variable name for the instruction parameter e g FlowDirection This variable allows you to turn the position of the valve to the direction you want Variable name definition Variable name Flow Direction C
3. 1 Switch on the AKTAdesign instrument with the mains switch located at the front left on the system base 2 Check that the computer and printer are switched on 3 Log in see the instrument user manuals 4 In UNICORN Main Menu Manager select Administration System Setup 5 Select System and then click Edit Click Component 6 From the Component list select the optional flow direction valve you have installed by checking the box or by clicking ADD 7 Click OK twice and then Close 8 Cut and mount the required PEEK capillaries Use a capillary with the correct i d as required by your system The principle flow path through the valve is shown below Reversed flow configuration DOWNWARD FLOW position 1 UPWARD FLOW position 2 To UV monitor Reverse flow Kit User Manual 71 5029 22 Edition AC 19 20 6 Installation Reverse flow Kit User Manual 71 5029 22 Edition AC Operation 7 7 Operation 7 1 Preparing for operation 1 A connected valve is automatically recognised by UNICORN at system start up Check that the added valve is indicated in the flow scheme in UNICORN 2 Verify that the valve is functioning properly by issuing manual commands from UNICORN as follows e n System Control select Manual FlowPath e Select instruction Valve7 AKTAreic FlowDirection valve AKTApurifier AKTAbasic e Select and execute a new position for the valve View the valve in the flow scheme panel to confirm tha
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5. B Bjorkgatan 30 SE 751 84 Uppsala Sweden GE Healthcare Europe GmbH Munzinger Strasse 5 D 79111 Freiburg Germany GE Healthcare UK Ltd Amersham Place Little Chalfont Bucki GE Healthcare Bio Sciences Corp 800 Centennial Avenue P O Box 1327 GE Healthcare Bio Sciences KK Nnghamhire HP7 9NA UK Piscataway NJ 08855 1327 USA Sanken Bldg 3 25 1 Huakunincho Shinjuku ku Tokyo 169 0073 Japan Asia Pacific Tel 852 2811 8693 Fax 852 2811 5251 e Australasia Tel 61 2 9899 0999 Fax 61 2 9899 7511 Austria Tel 01 57606 1619 Fax 01 57606 1627 e Belgium Tel 0800 73 888 Fax 03 272 1637 Canada Tel 800 463 5800 Fax 800 567 1008 e Central East amp South East Europe Tel 43 1 982 3826 Fax 43 1 985 8327 e Denmark Tel 45 16 2400 Fax 45 16 2424 e Finland amp Baltics Tel 358 0 9 512 39 40 Fax 358 0 9 512 39 439 e France Tel 01 69 35 67 00 Fax 01 69 41 96 77 e Germany Tel 0761 4903 490 Fax 0761 4903 405 e Italy Tel 02 27322 1 Fax 02 27302 212 Japan Tel 81 3 5331 9336 Fax 81 3 5331 9370 e Latin America Tel 55 11 3933 7300 Fax 55 11 3933 7304 e Middle East amp Africa Tel 30 210 9600 687 Fax 30 210 9600 693 Netherlands Tel 0165 580 410 Fax 0165 580 401 Norway Tel 815 65 555 Fax 815 65 666 Portugal Tel 21 417 7035 Fax 21 417 3184 e Russia amp other C I S amp N I S Tel 7 095 232 0250 956 1137 Fax 7 095 230 6377 South East Asia Tel 60 3 8024 2080 Fax 60 3 8024 20
6. GE Healthcare AKTAdesign Reverse flow Kit User Manual O AKTA Important user information All users must read this entire manual to fully understand the safe use of Reverse flow Kit WARNING The WARNING sign highlights instructions that A N must be followed to avoid personal injury It is important not to proceed until all stated conditions are met and clearly understood CAUTION The Caution sign highlights instructions that must be followed to avoid damage to the product or other equipment It is important not to proceed until all stated conditions are met and clearly understood Note The Note sign is used to indicate information important for trouble free and optimal use of the product CE Certifying This product meets the requirements of applicable CE directives A copy of the corresponding Declaration of Conformity is available on request The CE symbol and corresponding declaration of conformity is valid for the instrument when it is used as a stand alone unit or connected to other CE marked GE Healthcare instruments or connected to other products recommended or described in this manual and used in the same state as it was delivered from GE Healthcare except for alterations described in this manual Recycling This symobl indicates that the waste of electrical and electronic equipment must not be disposed as unsorted municipal waste and must
7. ancel Cear Help Click OK The defined variable is inserted in the method Return to the Variables page by selecting View Run setup to set the variable FlowDirection to the desired position Save the method Repeat step x y above to insert an instruction FlowDirection downflow This instruction should be inserted in instruction block Start_conditions This will ensure the flow returns to the Downflow position for the new purification run 7 6 AKTApurifier AKTAbasic UNICORN 4 12 7 6 1 Using the flow direction function in a run The flow direction valve instruction is designated FlowDirection in the flow scheme In the Run Data pane it is designated V7_FlowDir To use the flow direction valve in a method 1 In the Method Editor select File Method Wizard and start creating a new method In the Elution dialog select Segment Gradient Advanced under Elution Technique Click OK twice Select DownFlow or UpFlow under Flow Direction Save the method when finished Reverse flow Kit User Manual 71 5029 22 Edition AC 29 7 Operation 7 7 AKTApurifier UNICORN 5 10 7 7 1 Using the flow direction function in a run The flow direction valve instruction is designated FlowDirection in the flow scheme In the Run Data pane it is designated V7_FlowDir To use the flow direction valve in a method 1 In the Method Editor select File Method Wizard and start creating a new method 2 Inthe Elution dialog select Se
8. asic UNICORN 4 12 7 6 1 Using the flow direction function in a run 7 7 AKTApurifier UNICORN 5 10 ween 7 7 1 Using the flow direction function in a run Reverse flow Kit User Manual 71 5029 22 Edition AC 5 Contents Reverse flow Kit User Manual 71 5029 22 Edition AC Introduction 1 1 Introduction 1 1 General AKTAdesign standard system configurations can be changed to optional configurations This flexibility in AKTAdesign system strategies allows the user to enhance already used purification methods and also to develop new more complex methods When optional equipment is connected to a standard chromatographic system new sets of instructions to control the optional equipment become accessible for the user Optional configurations are selected installed and put into operation by the user An optional configuration consists of both hardware components and software instructions Optional configurations are monitored and controlled via methods run by the UNICORN control system in the same way as AKTAdesign standardsystem configurations 1 2 Optional flow direction valve V7 Preferentially in affinity based protein purification elution is carried out in the reversed direction to sample application This prevents strongly bound substances accumulated at the inlet side from interacting with the immobilized ligand throughout the column before elution and a dilution of the sample by eluting through the tota
9. be collected separately Please contact an authorized ME epresentative of the manufacturer for information concerning the decommissioning of equipment WARNING This is a Class A product In a domestic environment this product may cause radio interference in which case the user may be required to make adequate measures Contents Contents 1 Introduction Li SG SOR ccs chastised Meenas aaaceussans oo eeuiantereetastnte aed 7 1 2 Optional flow direction valve V7 vecceeccssssseesssecsceessssseeeecseeessssee 7 Safety ar SOEU an EARS 9 Unpacking Pre requisites Mounting of brackets Sb VaVe lD senaia A 16 5 2 UniNet Connection oe eesesseeeessssssssesessssssessceesnsseecessssseessesssneseeeesnnees 17 Installation 6 1 Preparation fOr USC un ssssseesssssscccssssssssseesscccssssssssseesccecsssssssnseeeseee 19 Operation 7 1 Preparing for operation s ssssssssssssssssssisssrsssrsssrsssressrsssrsrsrsssreeses 7 2 AKTArPLC UNICORN 3 24 3 21 ccccccccssssssssssscsssssssssssesetessssssee 7 2 1 Using the flow direction valve in a run i 7 3 AKTAFPLC UNICORN 4 04 4 12 nnii 7 3 1 Using the flow direction valve in a run eeccssssssssssssssssssssssssssssssseseeeeee 7 4 KTApurifier KTAbasic UNICORN 3 2 3 21 7 4 1 Using the flow direction function iN C run visseessssccscsssccsccceessessesees 7 5 AKTApurifier AKTAbasic UNICORN 4 0 4 11 7 5 1 Using the flow direction function in a method 7 6 AkTApurifier AKTAb
10. d via the UniNet 2 chain Note When Pump P 960 is used it is always installed as the last component in the UniNet 2 chain Since the pump has an internal termination no termination plug is needed Reverse flow Kit User Manual 71 5029 22 Edition AC Mounting of brackets 5 5 2 UniNet Connection Both the UniNet 1 and the UniNet 2 chain can be interrupted anywhere between the system pump and the termination plug to interconnect the optional components in the chain The termination plug can be moved to the last component furthest away from the system pump if motivated by cable routing considerations WARNING Only use mains cables delivered or approved by GE Healthcare To PC Controller board o O UniNet 2 Fraction chain collector e o Ts PC running UNICORN Termination plug UniNet 1 chain Use the attached UniNet 2 cables to connect the valve to the UniNet 2 communication link The valve can be connected anywhere between the system pump and the termination plug A N WARNING Do not block the rear panel of the system The mains power switch must always be easy to access Reverse flow Kit User Manual 71 5029 22 Edition AC 17 18 5 Mounting of brackets Reverse flow Kit User Manual 71 5029 22 Edition AC Installation 6 6 Installation 6 1 Preparation for use To start your AKTAdesign optional configuration system
11. eplace any item on the equipment during maintenance WARNING If there is a risk that large volumes of spilt liquid may penetrate the casing of the equipment and come into contact with the electrical components immediately switch off the chromatographic system and contact an authorized service technician WARNING When using hazardous chemicals take all suitable protective measures such as wearing protective glasses and gloves resistant to the chemicals used Follow local regulations and instructions for safe operation and maintenance of the system gt e BD CAUTION Make sure that the KTAdesign instrument is switched off before installing the optional components The mains power to the KTAdesign instrument must be switched OFF before disconnecting or connecting the UniNet 1 and UniNet 2 cables Reverse flow Kit User Manual 71 5029 22 Edition AC 10 2 Safety Reverse flow Kit User Manual 71 5029 22 Edition AC Unpacking 3 3 Unpacking Unpack the optional components and check against the supplied packing ist Inspect the items for obvious damage that may have occurred during transportation CAUTION Read the following information carefully as well as all the additional instructions supplied with the components to ensure that the AKTAdesign optional equipment is installed correctly Reverse flow Kit User Manual 71 5029 22 Edition AC 11 12 3 Unpacking Reverse flow Ki
12. fine a variable name for the instruction parameter e g Valve7_pos This variable allows you to turn the position of the valve to the port you want Variable name definition Variable name Valve _pos Cancel Clear Heb 7 Click OK The defined variable is inserted in the method 8 Return to the Variables page by selecting View Run setup to set the variable Valve7_pos to the desired position 9 Save the method 10 Repeat step x y above to insert an instruction Valve7 position 1 Downflow This instruction should be inserted in instruction block Start_conditions This will ensure the flow returns to the Downflow position for the new purification run Reverse flow Kit User Manual 71 5029 22 Edition AC 25 7 Operation 7 4 AKTApurifier AKTAbasic UNICORN 3 2 3 21 7 4 1 Using the flow direction function in a run The flow direction valve instruction is designated FlowDirection To use the valve in a method do as follows 1 Inthe Main Menu select File New Method to create a new method Select System Technique Template and For column Click OK 2 Select View Text instructions to display the text instruction editor Double click on the instruction block where you want to add the flow direction valve instruction e g Elution Highlight the elution instruction to insert the valve instruction Method Editor UNTITLED Bel File Block View Adviser Help dasa aw E Main 0 00 Base CY 0 10 Column ml
13. flow scheme and Run Data panes To use a valve in a method proceed as follows 1 Inthe Main Menu select File Method Wizard and create a new method 2 Select View Text instructions to display the text instruction editor Double click on the instruction block where you want to add the flow direction valve instruction e g Elution 3 Highlight the elution instruction Valve7 below to insert the valve instruction 4 Click the Flowpath radio button 5 From the Instructions list select valve Valve7 For this valve only two positions should be selected Position 1 Downflow position 2 Upflow A W U UU Block Start_Conditions Start_Conditions 0 00 Base SameAsMain 0 00 AveragingTimeUY 2 60 UV_Averaging_time W 0 00 Block Initial_Eluent_Conditions 0 00 End_block W 0 00 Block Column_Equilibration W 0 00 Block Flowthrough_Fractionation W 0 00 Block Sample_Injection W 0 00 Block Wash_Out_Unbound_Sample W 0 00 Block Eluate_and_Peak_Fractionation W 0 00 Block Gradient_Elution W 0 00 Block Clean_After_Gradient W 0 00 Block Reequilibration 0 00 End_method Click here to define a variable name for the instruction parameter Breakpoint p Instructions InjectionValve Position 1 8 Insert ao Hoy Fume w a I Feed ore a Flowpath a Var 7 AlamstMon InjectionMark Em Replace Delete Ready SS NOM i 24 Reverse flow Kit User Manual 71 5029 22 Edition AC Operation 7 6 De
14. gment Gradient Advanced under Elution Technique 3 Select DownFlow or UpFlow under Flow Direction 30 Reverse flow Kit User Manual 71 5029 22 Edition AC Operation 7 Method Wizard for System Purifier10 Gradient es8esaegaeass op os 10 15 20 25 30 35 40 45 so 4 Save the method when finished Reverse flow Kit User Manual 71 5029 22 Edition AC 31 www gehealthcare com lifescience GE Healthcare Bio Sciences AB Bj rkgatan 30 751 84 Uppsala Sweden Drop Design KTA AKTArpic AKTApuri trademarks of GE Healthcare compani monogram are trademarks of Genera Microsoft and Windows are either trad ier AKTAbasic and UNICORN are es GE imagination at work and GE Electric Company lemarks or registered trademarks of Microsoft Corporation in the United States and or other countries All goods and services are sold subjec to the terms and conditions of sale of the company within GE Healthcare which supplies them GE Healthcare reserves the right subject to any regu required to make changes in specifica latory and contractual approval if tions and features shown herein or discontinue the product described at any time without notice or obligation Contact your local GE Healthcare representative for the most current information 2005 General Electric Company Al rights reserved GE Healthcare Bio Sciences AB a General Electric Company GE Healthcare Bio Sciences A
15. l column volume Thus resulting peaks are sharper the substance elutes more concentrated and reversed elution speeds up the elution process Reverse flow Kit User Manual 71 5029 22 Edition AC 7 1 Introduction Flow Direction Valve V7 Flow restrictor Column On line filter Mixer Injection L Valve V1 Fraction A Pump A Pump B collector optional Outlet Valve V4 AKTApurifier AKTAbasic The optional flow direction valve V7 code no 18 1108 40 is available for e AKTArpic with UNICORN Software 3 2 or higher e AKTApurifier with UNICORN Software 3 21 AA or higher e AKTAbasic with UNICORN Software 3 21 AA or higher Reverse flow Kit User Manual 71 5029 22 Edition AC Safety 2 2 Safety 2 1 Safety IMPORTANT AKTAdesign is intended for laboratory use only not for clinical or in vitro use or for diagnostic purposes e The components are designed for indoor use only e Do not use in a dusty atmosphere or close to spraying water e Operate in accordance with local safety instructions WARNING When using hazardous chemicals all suitable protective measures such as protective glasses must be taken WARNING Ensure that the entire chromatographic system has been flushed thoroughly with distilled water before removing any capillaries or components WARNING Always disconnect the power supply before attempting to r
16. lock Clean_After_Gradient 0 00 Block Reequilibration 0 00 End_method CECA CACA CACA Click here to define a variable name for the instruction parameter Breakpoint i Injection al Position 1 8 a C Pump injectionValve 1 8 Insert Ie z I Feed Flowpath Var Change a C Alams amp Mon InjectionMark Chane Frac Replace C Watch C Other Delete Ready NUM 3 Click the Flowpath radio button 4 From the Instructions list select valve Valve7 For this valve only two positions should be selected Position 1 Downflow position 2 Upflow 5 Define a variable name for the instruction parameter Valve7_pos This variable allows you to turn the position of the valve to the port you want Variable name Valve7_pos Cancel Clear Help 6 Click OK The defined variable is inserted in the method 7 Return to the Variables page by selecting View Run setup to set the variable Valve7_pos to the desired position Reverse flow Kit User Manual 71 5029 22 Edition AC 23 7 Operation 8 Save the method 9 Repeat step x y above to insert an instruction Valve7 position 1 Downflow This instruction should be inserted in instruction block Start_conditions This will ensure the flow returns to the Downflow position for the new purification run 7 3 AKTAreic UNICORN 4 0 4 12 7 3 1 Using the flow direction valve in a run Flow direction valve instruction is designated Valve7 in the
17. quipment it is important to choose a location which minimize the length of the capillaries used to connect the optional components to the rest of the system Many components that are attached to the mounting rails uses a snap in bracket The bracket is supplied separately with the component and needs to be fitted as shown below before the component can be attached 2 Fit the attached mounting bracket with the two screws Inserting components lt N STL 1 Loosen the two screws Removing components Reverse flow Kit User Manual 71 5029 22 Edition AC 15 5 Mounting of brackets 5 1 Valve ID The flow direction valve has a unique ID code which identifies it to the UNICORN control system at system start up This ID code should be checked before installation The ID code for the flow direction valve and the corresponding instructions used in UNICORN to control it are as follows ID code Valve type Instruction in UNICORN 7 INV 907 Valve7 Components are connected to the UNICORN control system using eitherUniNet 1 or UniNet 2 cables Both the UniNet 1 and the UniNet 2 data communication chain in standard configuration is routed from the rear of the system pump via their respective components to the last component in the chain where it is terminated with a plug Frac 920 Frac 950 A 900 and A 905 are connected via the UniNet 1 chain All other optional components are connecte
18. t User Manual 71 5029 22 Edition AC Pre requisites 4 4 Pre requisites The general procedures for creating and editing methods are described in the UNICORN User Manuals Note The system configuration delay volume must be recalculated and reset when the standard configuration after the UV flow cell is changed AKTArpic AKTApurifier and AKTAbasic respectively must be installed and fully tested before the optional components are installed See the Installation Guide for your AKTAdesign chromatography system WARNING Ensure that the entire system has been flushed thoroughly with distilled water before removing any capillaries or components CAUTION Make sure that the AKTAdesign instrument is switched off before installing the optional components The mains power to the AKTAdesign instrument must be switched OFF before disconnecting or connecting the UniNet 1 and UniNet 2 cables Reverse flow Kit User Manual 71 5029 22 Edition AC 13 14 4 Pre requisites Reverse flow Kit User Manual 71 5029 22 Edition AC Mounting of brackets 5 5 Mounting of brackets CAUTION Make sure that the AKTAdesign instrument is switched off before installing the optional components The mains power to the AKTAdesign instrument must be switched OFF before disconnecting or connecting the UniNet 1 and UniNet 2 cables Optional equipment can be mounted on the system When selecting mounting location for the optional e
19. t the position change has occured Reverse flow Kit User Manual 71 5029 22 Edition AC 21 22 7 Operation 7 2 AKTArpic UNICORN 3 2 3 21 7 2 1 Using the flow direction valve in a run Flow direction valve instruction is designated Valve7 in the flow scheme and Run Data panes To use a valve in a method proceed as follows 1 Inthe Main Menu select File New Method to create a new method Select System Technique Template and For columni Click OK New Method Manual Injection FP Isocratic Manual Injection FP Segmented Manual Injection FP Step 2 Select View Text instructions to display the text instruction editor Double click on the instruction block where you want to add the general Reverse flow Kit User Manual 71 5029 22 Edition AC Operation 7 function valve instruction e g Gradient_Elution Highlight the elution instruction below to insert the valve instruction Method Editor UNTITLED Bl E File Block View Adviser Help Dele Sia fs 5 Bl E Main 0 00 Base CY 0 10 Column ml Any E W 0 00 Block Start_Conditions Start_Conditions 0 00 Base SameAsMain 0 00 AveragingTimeUY 2 60 UV_Averaging_time W 0 00 Block Initial_Eluent_Conditions 0 00 End_block W 0 00 Block Column_Equilibration 0 00 Block Flowthrough_Fractionation W 0 00 Block Sample_Injection E 0 00 Block Wash_Out_Unbound_Sample W 0 00 Block Eluate_and_Peak_Fractionation W 0 00 Block Gradient_Elution a 0 00 B
20. want Variable name definition Variable name ae e Clee Heb 6 Click OK The defined variable is inserted in the method 7 Return to the Variables page by selecting View Run setup to set the variable FlowDirection to the desired position 8 Save the method 9 Repeat step x y above to insert an instruction FlowDirection downflow This instruction should be inserted in instruction block Start_conditions This will ensure the flow returns to the Downflow position for the new purification run Reverse flow Kit User Manual 71 5029 22 Edition AC 27 28 7 Operation 7 5 AKTApurifier AKTAbasic UNICORN 4 0 4 11 7 5 1 Using the flow direction function in a method The flow direction valve instruction is designated FlowDirection in the flow scheme In the Run Data pane it is designated V7_FlowDir To use the flow direction valve in a method do as follows 1 Inthe Method Editor select File Method Wizard and create a new method 2 Select View Text instructions to display the text instruction editor Double click on the instruction block where you want to add the flow direction valve instruction e g Elution Highlight the elution instruction to insert the valve instruction amp Method Editor UNTITLED Bee File Block View Adviser Help Diels Sia lS Bl E Main 0 00 Base CV 0 10 Column ml Any 0 00 Block Start_Conditions Start_Conditions 0 00 Base SameAsMain 0 00 Wavelength 280 Wavelength_
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