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Akta FPLC instructions ENG

1. 5 Maintenance 5 4 Component maintenance The system documentation also includes a spare part list to be used to find common spare parts and their code numbers for ordering This list can also be found online at www gelifesciences com AKTA 5 5 Disassembly and assembly of components and consumables The operator must carefully read and understand the instructions supplied for each component before disassembly and assembly of the component When replacing consumables such as tubing and tubing connectors all neccessary safety precautions must be taken Contact your local GE Healthcare representative if further information or help is needed WARNING Disconnect power Always disconnect power from the instrument before replacing any component on the instrument unless stated otherwise in the user documentation WARNING Before disassembly check that there is no pressure in the piping system WARNING After assembly the piping system must be tested for leakage at maximum pressure for continued protection against injury risks due to fluid jets burst pipes or explosive atmosphere gt gt gt 5 6 Replacement of fuses WARNING Disconnect power Always disconnect power from the instrument before replacing fuses gt Refer to Section 7 1 Specifications on page 59 for information about the fuse type and rating If a fuse repeatedly blows switch off the system mains switch and contact yo
2. Figure 1 2 Basic flow path AKTAexplorer is shown as an example AKTAexplorer AKTApurifier and AKTAmicro Operating Instructions 28 9597 57 AA 11 Description Buffers from container pass through Buffer valve V6 and or the switch valve Sample pass through sample valve V5 which selects a sample depending on the setting in the control software Pump A pumps buffer through the system The buffers pass through Mixer M where they are mixed This is where the sample is added to the flow path The sample can be added manually with a suitable syringe or pumped in via the sample pump p1 from a sample chosen via sample valve V5 or an autosampler The flow direction valve V7 is optional in some systems and used to select the direction of the flow through column Column selection valves V2 and V3 direct flow through a specified column Liquid returns to flow direction valve V7 and redirects flow to outlet valve via pH UV and Condictivity monitors The outlet valve directs the flow either to waste to fraction col lection containers or to a fraction collector device such as Frac 950 Part pA V1 pl V7 V2 V3 V7 UV C pH V4 Step The flow path is likely to include air sensors flow restrictors online filters sample loops and so on which vary in number and application depending on the system and its strategy Fore more detai
3. OD xxxxx Sample Code No XXXXXXXX Voltage Serial No XXXXXXX Frequency Mfg Year 2009 Power max Fuse Qoy C C ULL LLLI Made in Sweden GE Healthcare Bio Sciences AB Or 9 751 84 Uppsala Sweden Symbols used in safety labels Label Meaning Warning Read the user documentation before using the system Do not N open any covers or replace parts unless specifically stated in the user documentation The system complies with the requirements for electromagnetic compliance EMC in Australia and New Zealand The system complies with applicable European directives 22 KTAexplorer AKTApurifier and AKTAmicro Operating Instructions 28 9597 57 AA 2 Safety instructions 2 2 Labels Labels concerning hazardous substances Label Meaning This symbol indicates that the waste of electrical and electronic equipment must not be disposed as unsorted municipal waste and must be collected separately Please contact an authorized representative of the manufac mummm turer for information concerning the decommissioning of equipment This symbol indicates that the product contains hazardous materials in o excess of the limits established by the Chinese standard SJ T11363 2006 Requirements for Concentration Limits for Certain Hazardous Substances in Electronics 2 5 Emergency procedures This section describes how to do an emergency shutdown of the AKTAexplorer AKTApurifier and A
4. 4 Operation 4 5 Setting up a run 10 In the Save As dialog e Enter method name in the Method name field and select folder to save the method in e Ifyou have more than one system connected to the computer select system in the For System area for which the method is intended Select technique in the Technique area for which the method was written 11 Click OK The method is saved It can now be started from the System Control module 4 6 Performing a run 1 Select method a In System Control select File Run b Select the required Method from the list 2 Specify variables Enter identification names for the samples via the keyboard 3 Editresult file location and names If required edit the folder path and file names of the result files to be created 4 Preparations completed Ensure that the preparations according to Section 4 4 Preparations before start on page 32 has been performed 5 Check the flow path Make sure that e there is enough buffer available e the correct inlet is placed in each buffer e the outlets are placed in correct bottles e the columns are placed in correct positions e the chosen fraction collector rack is filled and is in correct place 6 Prepare the samples The samples should have been prepared and clarified using centrifugation and or filtration through a 0 45 um filtert 1 fusing HisTrap FF crude clarification is not needed AKTAexplorer AKTApurifier and AKTAmicro
5. 2 Safety instructions 2 1 Safety precautions WARNING Spread of biological agents The operator has to take all necessary actions to avoid spreading hazardous biological agents in the vicinity of the instrument The facility should comply with the national code of practice for biosafety WARNING High pressure The AKTAexplorer AKTApurifier and AKTAmicro instrument operates under high pressure Wear protective glasses at all times Installing and moving the instrument WARNING Supply voltage Make sure that the supply voltage at the wall outlet corresponds to the marking on the instrument before connecting the power cord WARNING Protective ground The AKTAexplorer AKTApurifier and AKTAmicro instrument must always be connected to a grounded power outlet WARNING Power cord Only use power cords delivered or approved by GE Healthcare AKTAexplorer AKTApurifier and AKTAmicro Operating Instructions 28 9597 57 AA 17 WARNING Access to power switch and power cord Do not block the rear and side panel of the instrument The Power switch must always be easy to access The power cord must always be easy to disconnect WARNING Installing the computer The computer should be installed and used according to the instructions provided by the manufacturer of the computer CAUTION Heavy object Use suitable lifting equipment when moving the systems Three people are requ
6. Using flammable liquids e Personal protection Installing and moving the instrument e System operation e Maintenance General precautions Always follow these General precautions to avoid injury when using the AKTAexplorer AKTApurifier and AKTAmicro instrument WARNING Do not operate the AKTAexplorer AKTApurifier and AKTAmicro instrument in any other way than described in the AKTAexplorer AKTApurifier and AKTAmicro and UNICORN manuals 14 AKTAexplorer AKTApurifier and AKTAmicro Operating Instructions 28 9597 57 AA 2 Safety instructions 2 1 Safety precautions WARNING Operation and user maintenance of the AKTAexplorer AKTApurifier and AKTAmicro instrument should be performed by properly trained personnel only WARNING Before connecting a column to the AKTAexplorer AKTApurifier and AKTAmicro instrument read the instructions for use of the column To avoid exposing the column to excessive pressure make sure that the pressure limit is set to the specified maximum pressure of the column WARNING Do not use any accessories not supplied or recommended by GE Healthcare WARNING Do not use the AKTAexplorer AKTApurifier and AKTAmicro instrument if it is not working properly nor if it has suffered any damage for example PP gt gt e damage to the power cord or its plug e damage caused by dropping the equipment e damage caused by splashing liquid onto it CAUTION Waste tubes
7. The AKTAexplorer AKTApurifier and AKTAmicro systems are intended for research use only and shall not be used in any clinical procedures or for diagnostic purposes Safety notices This user documentation contains WARNINGS CAUTIONS and NOTICES concerning the safe use of the product See definitions below Warnings WARNING WARNING indicates a hazardous situation which if not avoided could result in death or serious injury It is important not to proceed until all stated conditions are met and clearly understood 6 AKTAexplorer AKTApurifier and AKTAmicro Operating Instructions 28 9597 57 AA 1 Introduction 1 1 Important user information Cautions CAUTION CAUTION indicates a hazardous situation which if not avoided could result in minor or moderate injury It is important not to proceed until all stated conditions are met and clearly understood Notices NOTICE 9 NOTICE indicates instructions that must be followed to avoid damage to the product or other equipment Notes and tips Note A Note is used to indicate information that is important for trouble free and optimal use of the product TIP A tip contains useful information that can improve or optimize your procedures Typographical conventions Software items are identified in the text by bold italic text A colon separates menu levels thus File Open refers to the Open command in the File menu Hardware items are identified in the text
8. 2 Safety instructions 2 1 Safety precautions 20 2 Safety instructions 2 1 Safety precautions WARNING After assembly the piping system must be tested for leakage at maximum pressure for continued protection against injury risks due to fluid jets burst pipes or explosive atmosphere WARNING Before disassembly check that there is no pressure in the piping system WARNING Disconnect power Always disconnect power from the instrument before replacing fuses WARNING Decontaminate the equipment before decommissioning to ensure that hazardous residues are removed NOTICE Cleaning Keep the instrument dry and clean Wipe regularly with a soft damp tissue and if necessary a mild cleaning agent Let the instrument dry completely before use o gt lt gt gt 2 2 Labels This section describes the safety labels and labels concerning hazardous substances that are attached to the AKTAexplorer AKTApurifier and AKTAmicro instruments For information about marking of the computer equipment refer to the manufacturer s instructions AKTAexplorer AKTApurifier and AKTAmicro Operating Instructions 28 9597 57 AA 21 2 Safety instructions 2 2 Labels Labels on the instrument The illustration below shows an example of the identification label that is attached to the AKTAexplorer AKTApurifier and AKTAmicro instruments S
9. AKTAmicro User Manuals 53 Possible cause Aging UV lamp UV lamp in wrong po sition The theoretical extinc tion coefficient too low 6 2 Conductivity curve problems Possible cause Air in the pump or the flow cell Leaking tube connec tions Incorrect mixer func tion Dirty conductivity cell Column not equilibrat ed KTAexplorer KTApurifier and AKTAmicro Operating Instructions 28 9597 57 AA Error symptom Low sensitivity Error symptom Baseline drift or noisy signal Corrective action Clean the flow cell according to procedure in Monitor User Manual Use a temperature compensation factor See Monitor User Manual Check that the pump is operating and is programmed correctly Check that the mixing chamber is free from dirt or particles Change to a larger mixing cham ber volume if necessary Check the motor operation Place a hand on the mixer and start it by starting the pump at a low flow rate You should both hear and feel the mixer motor and stirrer when they are spinning Check for loose tubing connec tions Use the flow restrictor Wash the tubing and check pump is operating properly Change to smaller mixer volume AKTAexplorer AKTApurifier and AKTAmicro Operating Instructions 28 9597 57 AA Possible cause Dirty flow cell Decrease in ambient temperature Incorrect pump func tion Dirty mixing chambe
10. three people to lift and move it unless a suitable lifting device is used AKTAexplorer 66 kg AKTApurifier 41 kg AKTAmicro 55 kg The instrument can be transported on a trolley or a suitable lifting device capable of supporting the weight of the instrument AKTAexplorer AKTApurifier and AKTAmicro Operating Instructions 28 9597 57 AA 26 3 Installation 3 2 Transport NOTICE o Lift the system in the upright position Do not use the front panel bar as a lifting handle Before moving the system e disconnect all cables and tubing connected to peripheral components and liquid containers remove all items from the top of the system e close the valve door completely only for AKTAexplorer e grasp the system firmly by placing the fingers in the gap between the swivel platform and the base of the main unit and lift For more information on Transport see AKTAexplorer AKTApurifier and AKTAmicro Installation Guides 5 5 Unpacking e Check the equipment for damage before starting assembly and installation e Document any damage and contact your local GE Healthcare representative Remove straps and packing material and stand equipment upright on swivel foot before starting installation 5 4 Assembly The following parts must be added to the AKTAexplorer AKTApurifier and AKTAmicro instruments before they can be used pH electrode optional Waste tube e CU 950 Cont
11. B2 Teflon 1700 16 3 4 x 10 Switch valve pump A3 B3 Teflon 150 1 6 302 Pump Mixer G1 G2 PEEK 330 0 50 65 Mixer On line filter G3 PEEK 200 0 50 39 Filter V1 Inj Valve G4 PEEK 180 0 50 35 V1 UV bypass G5 PEEK 400 0 50 79 UV Cond Cell G6 PEEK 160 0 25 8 Cond Cell Flow restrictor G7 PEEK 80 0 25 4 Flow restrictor V4 G8 PEEK 140 0 25 7 V4 Fraction collector G9 PEEK 600 0 25 29 V4 Flow through F3 PEEK 1000 0 50 196 V1 V4 Waste W1 W2 W3 Tefzel 100 0 75 442 AKTAexplorer AKTApurifier and AKTAmicro Operating Instructions 28 9597 57 AA 1 6 Tubing Table 6 4 Tubing specifications for AKTApurifier 100 Use Label Material Length I D Volume mm mm ul Inlet A1 Al Teflon 1700 29 112 x 10 Inlet A2 A2 Teflon 1900 9 12 5 x 103 Inlet B1 B1 Teflon 1500 29 9 9 x 10 Inlet B2 B2 Teflon 1700 29 112 x 10 Switch valve pump A3 B3 Teflon 150 2 9 991 Pump Mixer G1 G2 PEEK 330 0 75 146 Mixer On line filter G3 PEEK 200 0 75 88 Filter V1 Inj Valve G4 PEEK 180 0 75 80 V1 UV bypass G5 PEEK 400 0 75 177 UV Cond Cell G6 Tefzel 160 0 75 71 Cond Cell Flow restrictor G7 Tefzel 80 0 75 35 Flow restrictor V4 G8 Tefzel 140 0 75 62 V4 Fraction collector G9 Tefzel 600 0 75 265 V4 Flow through F3 Tefzel 1000 1 0 785 V1 V4 Waste W1 W2 W3 Tefzel 100 0 75 442 Tubing specifications for AKTAmicro Names in the Tubing i
12. Replace the membranes and piston seals according to the User manual AKTAexplorer AKTApurifier and AKTAmicro Operating Instructions 28 9597 57 AA 45 Instructions reference Disconnect the flow restrictor Connect a capillary approx 1 m i d 1 mm to a free port in the injection valve Set the valve manually to this port Put the open end in a waste container Run the pump at 10 ml min with water Note the back pressure Bp1 on the pump display or in the Run Data window Connect the flow restrictor to the open end of the capillary observe the IN marking Putthe flow restrictor in the waste contain er Run the pump at 10 ml min with water Note the back pressure Bp2 on the pump display or in the Run Data window Calculate the back pressure generated by the flow restrictor Replace it if it is not within limit Check that flow restrictor generates the follow ing back pressure FR 904 0 4 0 05 MPa FR 902 0 2 x0 05 MPa Check the back pressure as follows 1 2 Action Flow restrictor AKTAexplorer AKTApurifier and AKTAmicro Operating Instructions 28 9597 57 AA 5 Maintenance 5 2 User maintenance schedule Interval Monthly 46 5 Maintenance 5 2 User maintenance schedule Interval Action Instructions reference 1 Disconnect the flow restrictor 2 Connecta capillary approx 1 m i d 1 mm to the waste port port 5 on the injection valve Set the injec
13. User Manual Ifthe problem remains replace the pH electrode 55 Possible cause Loose connection of conductivity flow ca ble Incorrect pump and valves function Incorrect temperature compensation factor Dirty or incorrectly equilibrated column Incorrect mixer func tion Possible cause Faulty electrode con nection Damaged electrode Incorrectly connected pH monitor Dirty pH electrode KTAexplorer KTApurifier and AKTAmicro Operating Instructions 28 9597 57 AA Error symptom Incorrect or unstable reading 6 3 pH curve problems Error symptom No response to pH changes Small response to pH changes 6 Troubleshooting 6 3 pH curve problems Error symptom Possible cause Corrective action Slow pH responseor Contaminated elec Check the electrode glass mem Calibration impossible trode glass mem brane If it is contaminated clean brane the electrode following the instruc tions in Monitor pH C 900 User Manudl or UPC 900 User Manudl Membrane has dried Ifthe membrane has dried out the out electrode may be restored by soaking it in buffer overnight 56 AKTAexplorer AKTApurifier and AKTAmicro Operating Instructions 28 9597 57 AA 6 Troubleshooting 6 5 pH curve problems Error symptom Possible cause Corrective action Incorrect or unstable Problem with elec Check that the electrode cable is pH reading trode conn
14. by bold text e g Power switch 1 2 Regulatory information This section describes the directives and standards that are fulfilled by the AKTAexplorer AKTApurifier and AKTAmicro systems AKTAexplorer AKTApurifier and AKTAmicro Operating Instructions 28 9597 57 AA 7 1 Introduction 1 2 Regulatory information Manufacturing information The Declaration of Conformity includes the following information Requirement Content Name and address of manufacturer GE Healthcare Bio Sciences AB Bj rkgatan 30 SE 751 84 Uppsala Swe den Name and address of person responsible Peter L wendahl Bj rkgatan 30 for Technical File SE 751 84 Uppsala Sweden Name and ID of notified body INTERTEK SEMKO AB NB 0413 Place and date of declaration Uppsala Sweden May 2009 Identity of person authorizedto sign DoC Peter L wendahl See EC Declaration of Conformity CE Conformity This product complies with the European directives listed in the table by fulfilling the corresponding harmonized standards A copy of the Declaration of Conformity is available on request 2006 42 EC Machinery Directive MD 2006 95 EC Low Voltage Directive LVD 2004 108 EC ElectroMagnetic Compatibility EMC Directive International standards This product fulfills the requirements of the following standards Standard Description EN 61010 1 IEC Safety requirements for electrical 61010 1 UL 61010 1 equipment f
15. d column in Table refer to tubing labels in the liquid flow path connection diagram see Liquid flow path for AKTAmicro on page 68 Tubing Material Color Max Volume Connection points o d pressure of 10cm 0 35 mm 1 6 Titanium Grey 35MPa 9 6 ul From Pump P 905 to Mixer G1 G2 mm 72 AKTAexplorer AKTApurifier and AKTAmicro Operating Instructions 28 9597 57 AA 6 Tubing Tubing Material Color Max Volume Connection points o d pressure of 10 cm Union PEEK Black gt 35 MPa Between Mixer and On line m m filter Between UV cell and Conduc tivity cell 0 15 mm 1 16 PEEK Violet gt 35MPa 18yl From On line filter to outlet G3 to or to fraction collector MS G5 optional PEEK tubing i d 0 15 mm is installed at delivery 0 15 mm 375 Fused sili Brown 35 MPa 18yl From On line filter to outlet Di 0 15 um ca or to fraction collector MS optional Tubing kit 0 15 is supplied with the system 0 10 mm 200 Fused sili Brown 35 MPa 0 8 ul From On line filter to outlet Di 0 10 um ca or to fraction collector MS optional Tubing kit 0 10 is supplied with the system 0 75 mm 1 16 Tefzel Clear 7 MPa 44 2 ul Waste tubing w1 W2 1 6 mm 1 8 Teflon Clear 3 4 MPa 201 1 ul Inlet tubing to Pump P 905 A1 to A3 B1 to B3 1l 025and0 5 mm i d tubing is supplied for non analytical applications AKTAexplorer A
16. degree of contamination depends on the sample and the chromatographic conditions employed These should be considered when designing a cleaning protocol Routine cleaning should be performed at intervals aimed at prevention rather than cleaning the system from growth or contamination WARNING Make sure that the piping system is completely leakage free before performing any CIP on the system Make sure that the process control method for cleaning flushes all possible flow paths in the system After cleaning rinse the entire system with water or suitable liquid until the piping tubing system is completely free from the CIP solution monitors in the system can be used as detectors Do not leave NaOH or other cleaning agents in the system for long periods WARNING Hazardous chemicals during maintenance When using hazardous chemicals for system or column cleaning wash the system or columns with a neutral solution in the last phase or step WARNING NaOH is corrosive and therefore dangerous to health When using hazardous chemicals avoid spillage and wear protective glasses and other suitable personal protective equipment See also Section 5 8 Storage on page 50 5 4 Component maintenance Maintenance and preventive replacement of parts of the major components are described in the respective manuals included in the system documentation 48 AKTAexplorer AKTApurifier and AKTAmicro Operating Instructions 28 9597 57 AA
17. of operations and length of intervals necessary to maintain system function and safety Table 5 1 User maintenance schedule Instructions reference Visually inspect the system for leaks 1 Forcleaning the flow path see Cleaning In Place on page 48 2 For leaving the system for a few days see Section 5 8 Storage on page 50 Calibrate the pH electrode if applicable accord ing to Monitor pH C 900 User Manual Action Leak inspection Wash the system flow path Calibrate pH electrode optional AKTAexplorer AKTApurifier and AKTAmicro Operating Instructions 28 9597 57 AA Interval Daily 44 5 Maintenance 5 2 User maintenance schedule Interval Action Instructions reference Weekly Check inlet filters Check the inlet filters visually and replace them if necessary Replace on line filter Replace the on line filter if applicable Change pump rinsing Change rinsing solution Always use 20 solution ethanol with 10 mM NaOH as rinsing solution If the volume of rinsing solution in the storage bottle has increased it can be an indication of internal pump leakage Replace the piston seals according to the User manual If the volume of rinsing solution in the storage bottle has decreased significantly check if the rinsing system connectors are mounted proper ly If the rinsing system connectors are not leaking the rinsing membranes or piston seals may be leaking
18. 0 on page 65 and Liquid flow path for AKTApurifier 10 and 100 on page 67 respectively Table C 1 Tubing specifications for AKTAexplorer 10 Use Label Material Length Volume mm mm ul Inlets A11 to A18 A11 18 Teflon 1250 1 6 2 5 x 10 Inlet A1 1 Teflon 750 1 6 1 5 x 10 Inlet A2 A2 Teflon 2000 1 6 4 0 x 10 Inlet 81 B1 Teflon 1800 16 3 6 x 10 Inlet B2 B2 Teflon 1800 16 3 6 x 10 Switch valve pump A3 B3 Teflon 150 1 6 302 Pump Mixer 61 G2 PEEK 300 0 50 59 Mixer On line filter G3 PEEK 150 0 50 29 Filter V1 Inj Valve G4 PEEK 460 0 50 90 V1 V2 G5 PEEK 270 0 25 13 V2 V3 bypass G6 PEEK 620 0 50 122 V3 UV G7 PEEK 550 0 25 27 UV Cond Cell G8 PEEK 160 0 25 8 Cond Cell Flow restrictor G9 PEEK 450 0 25 22 Flow restrictor V4 G10 PEEK 120 0 25 6 V4 Fraction collector G11 PEEK 600 0 25 29 V4 Flow through F3 PEEK 1000 0 50 196 V1 Waste W1 W2 Tefzel 1300 0 75 574 AkTAexplorer AKTApurifier and AKTAmicro Operating Instructions 28 9597 57 AA 69 6 Tubing Material Length I D mm mm V4 Waste W3 Tefzel 1000 0 75 442 Table C 2 Tubing specifications for AKTAexplorer 100 Material Length 1 D mm mm Inlets A11 to A18 A11 18 Teflon 1250 2 9 8 3 x 10 Inlet A1 Al Teflon 750 29 4 9 x 103 Inlet A2 A2 Teflon 2000 9 13 2 x 103 Inlet B1 B1 Teflon 1800 29 11 9 x 103 Inlet B2 B2 Tefl
19. 2125 02 9 7783 20 2 29340 81 6 67 68 5 75 08 1 Exposure up to 2 months OK OK Avoid OK OK OK OK OK OK OK OK Avoid OK OK Avoid Avoid OK AKTAexplorer AKTApurifier and AKTAmicro Operating Instructions 28 9597 57 AA 7 Reference information Exposure gt 1day OK OK OK OK OK OK OK OK OK OK OK OK OK OK Avoid Avoid OK 7 2 Chemical resistance Chemical Acetic acid 70 Acetonitrile Acetone 10 Ammonia 30 Ammonium chlo ride Ammonium bicar bonate Ammonium nitrate Ammonium sul phate 1 Butanol 2 Butanol Citric acid Chloroform Cyclohexane Detergents Dimethyl sulphox ide 1 4 Dioxane Ethanol 100 60 7 Reference information 7 2 Chemical resistance Comments Silicone not resis tant Pressure limit for PEEK decreases Silicone not resis tant Silicone not resis tant Pressure limit for PEEK decreases Silicone not resis tant Silicone not resis tant Titanium is af fected by long term use Silicone not resis tant Elgiloy is affected by long term use Titanium alumini um oxide and glass are affected by long term use 61 27 35 2 7 1 75 7 777 2 7 6 2 2 7 53 228 Chemical Exposure Exposure 1 day up to 2 months Ethyl acetate OK Avoid Ethylene glycol
20. Electrical shock hazard All repairs should be done by service personnel authorized by GE Healthcare Do not open any covers or replace parts unless specifically stated in the user documentation System operation o gt b b Maintenance AKTAexplorer AKTApurifier and AKTAmicro Operating Instructions 28 9597 57 AA WARNING Disconnect power Always disconnect power from the instrument before replacing any component on the instrument unless stated otherwise in the user documentation WARNING Hazardous chemicals during maintenance When using hazardous chemicals for system or column cleaning wash the system or columns with a neutral solution in the last phase or step WARNING Do not perform any type of maintenance work while the system is powered electrically or when the piping system is pressurized Note that the piping system can be pressurized even when the system is closed down WARNING Only spare parts that are approved or supplied by GE Healthcare may be used for maintaining or servicing the system WARNING Make sure that the piping system is completely leakage free before performing any CIP on the system WARNING NaOH is corrosive and therefore dangerous to health When using hazardous chemicals avoid spillage and wear protective glasses and other suitable personal protective equipment AKTAexplorer AKTApurifier and AKTAmicro Operating Instructions 28 9597 57 AA
21. GE Healthcare AKTAexplorer AKTApurifier and AKTAmicro Operating Instructions Original instructions 0 KTA Table of Contents Table of Contents 1 5 11 Important user information m 6 1 2 Regulatory information 7 13 Instrument a 10 T4 Control SOFtWOFe aceite ette eec d b tees tr p o OR HR ER 12 2 Safety WAST DONE auo adstat ioo ticc tortas ka lae tael cha aho iuda latius ctos TERETE Sacs 14 2 1 Safety precautions a 14 22 us 21 2 3 Emergency procedutes sui e baee ti e better oor cU cH 23 24 24 LE LC S ce ene ene nee sone Mn nan E er See ee re 26 3 1 SILETEGUIMEMMEMIUS ennha ect tu 00 26 3 2 E AE TES 26 AE 27 3 4 Assembly 27 3 5 Connections sss m 28 3 6 Spare parts and aecessories us eie ie I Hei hei po 29 4 Operation en 30 4 1 Operation overview 30 4 2 Starting the instrument 30 4 5 Starting the control system 31 A Preparations before SEGFt siae ten tetris retreat eret retia erue 32 4 5 7 35 4 6 Performing 39 4 7 Procedures after a run ttti ttt ttt 42 5 6 0 16 43 51 43 5 2 User maintenance schedule 44 5 anes 47 5 4 Component maintenance m 48 5 5 Disassembly and assembly of components and consumables ss 49 56 Replacement OF fusis enpuer t et RR bu asia N RC e
22. KTAmicro systems The section also describes the result in the event of power failure Emergency shutdown In an emergency situation do as follows to stop the run Step Action 1 To pause the run from UNICORN click the Pause button in System Control 2 If required switch off power to the instrument by pressing the Main power Switch to the 0 position AKTAexplorer AKTApurifier and AKTAmicro Operating Instructions 28 9597 57 AA 23 2 Safety instructions 2 5 Emergency procedures Power failure The result of a power failure depends on which unit that is affected Power failure to will result in AKTAexplorer AKTApuri The runis interrupted immediately in an undefined fier and AKTAmicro sys state tem gt The data collected up to the time of the power failure is available in UNICORN Computer e The UNICORN computer shuts down in an undefined state e The run continues but data cannot be saved in UNI CORN 2 4 Recycling information The equipment shall be decontaminated before decommissioning and all local regulations shall be followed with regard to scrapping of the equipment Disposal general instructions When taking AKTAexplorer AKTApurifier and AKTAmicro systems out of service the different materials must be separated and recycled according to national and local environmental regulations Recycling of hazardous substances AKTAexplorer AKTApurifier and AKTAmicro instrument
23. KTApurifier and AKTAmicro Operating Instructions 28 9597 57 AA 73 GE imagination at work and GE monogram are trademarks of General Electric Company Drop Design HisTrap Superloop UNICORN AKTA AKTAexplorer AKTAmicro and AKTApurifier are trademarks of GE Healthcare companies All third party trademarks are the property of their respective owners 2009 General Electric Company All rights reserved First published Jul 2009 All goods and services are sold subject to the terms and conditions of sale of the company within GE Healthcare which supplies them A copy of these terms and conditions is available on request Contact your local GE Healthcare representative for the most current information UNICORN Any use of this software is subject to GE Healthcare Standard Software End User License Agreement for Life Sciences Software Products GE Healthcare Europe GmbH Munzinger Strasse 5 D 79111 Freiburg Germany GE Healthcare UK Limited Amersham Place Little Chalfont Buckinghamshire HP7 9NA UK GE Healthcare Bio Sciences Corp 800 Centennial Avenue PO Box 1327 Piscataway NJ 08855 1327 USA GE Healthcare Japan Corporation Sanken Bldg 3 25 1 Hyakunincho Shinjuku ku Tokyo 169 0073 Japan 28 9597 57 AA 11 2009 For local office contact information visit www gelifesciences com contact GE Healthcare Bio Sciences AB Bj rkgatan 30 751 84 Uppsala Sweden www gelifesciences com AKTA imagi
24. M NaOH or 2096 ethanol solution Columns and media shall be stored according to their respective instructions AKTAexplorer AKTApurifier and AKTAmicro Operating Instructions 28 9597 57 AA 50 5 Maintenance 5 8 Storage Storage conditions The following conditions shall be maintained while the system is in storage Temperature 2 to 30 C preferably room temperature Relative humidity 0 to 95 non condensing preferably low humidity After storage clean the system calibrate all monitors and perform a leakage test before using the system AKTAexplorer AKTApurifier and AKTAmicro Operating Instructions 28 9597 57 AA 51 Corrective action Clean the system Make sure air is removed Clean the column according to the column instructions Check the mixer function by plac ing a stirrer bar on top of the mixer housing The stirrer bar should ro tate when the system is in Run mode The mixer function can also be checked by running the installa tion test Check the connections of the UV cell optical fiber Replace if neces sary Clean the UV cell by flushing De cone 90 Deconex 11 or equivalent Check if the signal is still noisy with water Purge the pump according to Pump User Manual Run a system wash with buffer AKTAexplorer AKTApurifier and AKTAmicro Operating Instructions 28 9597 57 AA 6 Troubleshooting 6 Troubleshooting Possible cause Dirt or resi
25. OK OK 107 21 1 100 Formic acid 100 OK OK 64 18 6 Glycerol 100 OK OK 56 81 5 Guanidinium hy OK OK drochloride Hexane OK Avoid Hydrochloric acid OK OK 7647 01 0 0 1 M Hydrochloric acid OK Avoid gt 0 1M Isopropanol 100 OK OK 67 63 0 Methanol 100 OK OK 74 93 1 Nitric acid diluted OK Avoid Nitric acid 30 Avoid Avoid Phosphoric acid OK Avoid 7664 38 2 10 Potassium carbon OK OK 584 08 7 ate Potassium chloride OK OK 7447 40 7 KTAexplorer KTApurifier and AKTAmicro Operating Instructions 28 9597 57 AA Exposure Comments up to 2 months Avoid ETFE PP and PE not resistant OK OK OK OK OK OK 7647 14 5 231 598 3 Avoid 11 2 2 5 PVDF and borosili cate glass are af fected by long term use OK 7757 82 6 231 820 9 Avoid PEEK and titanium are affected by long term use Avoid Avoid Silicone PP and PE are not resistant Avoid ETFE CTFE PP and PE are not resistant Avoid Pressure limit for PEEK decreases OK 76 03 9 200 927 2 OK 176 05 1 200 929 3 OK 57 13 6 200 315 5 AKTAexplorer AKTApurifier and AKTAmicro Operating Instructions 28 9597 57 AA 7 Reference information Exposure gt 1day Avoid OK OK OK OK OK OK OK OK OK Avoid Avoid Avoid OK OK OK OK 7 2 Chemical resistance Chemical Pyridine Sodium acetate Sodium bicarbon ate Sodium bisulph
26. Operating Instructions 28 9597 57 AA 39 4 Operation 4 6 Performing a run a Place the sample in chosen liquid container vials for the autosampler or fill the capillary loop or the Superloop with sample depending on chosen method Make sure that no air enters the tubing Place the tubing close to the bottom of the liquid container but not too tight against the bottom b Secure the tubing 7 Final check Perform a final check that tubing columns solutions and buffers are placed according to the method 8 Start the run Click START to start the run on the selected systems Viewing the run The progress of the run can be viewed in detail in the System Control module Method q System1 Wonbytare Capto Capto QYObM bsmax dead volume m01 Q System Control 1 System 1 Result q System1 Wonbytare Capto Capto QV bsmax001 fF Temp EN p 6 Controlled By Administrator SamplePres No watch Acc Time Block Time Pure sample File View Manual System Help Run Hold Pause Continue End 0 Instruments pn Run Status I Volume Block Volume mm uv Cond and Conc Pressure Flow TI nze s Port amp Port 8 Port S Port Dy y v OL OF OMG 9 Column Coltmn Q t et Buffer Salve Galve Valve corr Gradient TUTAB TUTT 8 00 ml Gradient 0 0 B 0 00 ml 321 ml Paus
27. RN Manager window see Figure 4 1 AKTAexplorer AKTApurifier and AKTAmicro Operating Instructions 28 9597 57 AA 35 4 Operation 4 5 Setting up a run The New Method dialog opens New Method 2 If several systems are available select which system to use in the For system list box 3 Select Wizard to create a method using the Method Wizard Click OK 36 AKTAexplorer AKTApurifier and AKTAmicro Operating Instructions 28 9597 57 AA 4 Operation 4 5 Setting up a run The Method Wizard dialog opens Method Wizard for System AA E10S Main Selection Column and BufferPrep Main Selection n a Column Any Column Position Position Bypass Flexible Flow Rates NOTE Different Flow Rates are Set on Variable Page C Flow Regulation of the System Pump BufferPrep 4 Inthe Method Wizard dialog e Select the appropriate chromatographic technique in the Main Selection list box for example Affinity Select the column you intend to use in the Column list box The correct column volume the recommended flow rate and the correct pressure limit for that column will then be automatically implemented in the method Select column position in the Column Position list box e If required select Flexible Flow Rates and or Flow Regulation of the System Pump and or BufferPrep 5 Click Next 6 Oneach new page in the Method Wizard select the appropriate parameters and click Next to continue AK
28. TAexplorer AKTApurifier and AKTAmicro Operating Instructions 28 9597 57 AA 37 4 Operation 4 5 Setting up a run 7 On the last page click Finish The Run Setup dialog opens with the Variables tab selected by default Evaluation Procedures Method Information Start Protocol Questions Result Name Frac 950 Variables Scouting Notes Gradent BufferPrep Columns Reference Curves Value Range HiTrap Protein amp HP 5 ml Global penser Tt Cabin Presse MPa T NN tart Inst pT vert 2 OFF ma _Sample_PressLimit PressLimit Im an j 00 1 00 na lil seme 3 eS pese Naini sna ae ee C Show details CI Show unused variables v Display tooltip for extended variable cells E dit Variable 8 The method is represented by a number of blocks on the Variables tab The blocks are typical steps in a chromatographic run Each block contains a number of method variables If necessary change the variables to suit your application 9 Inthe Method Editor select File Save As to save the method The Save As dialog opens Save As c SDefault RL ETUXT2 Technique ny e B Anion Exchange Cation Exchange RPC HIC Size Exclusion Affinity Method name Chromatofocusing 8 AKTAexplorer AKTApurifier and AKTAmicro Operating Instructions 28 9597 57 AA
29. a run 2 The New Method icon opens the Method Editor module and displays the New Method di alog box 3 The System Control icon activates the System Control module and displays the Manual instruction dialog box Figure 4 1 The UNICORN Manager window Control system in UNICORN To open the System Control module in UNICORN click the System Control icon in the UNICORN Manager window see Figure 4 1 4 4 Preparations before start Prepare buffers solutions and inlets 1 Prepare buffers and solutions required for the run 32 AKTAexplorer AKTApurifier and AKTAmicro Operating Instructions 28 9597 57 AA 4 Operation 4 4 Preparations before start 2 Immerse all inlet tubing in the appropriate liquid containers as described in the method Purging the pump and inlet tubing Fill the pump and inlet tubing with liquid if small amounts of air need to be removed or if the inlet tubing is empty To fill the inlet tubing manually in System Control 1 Make sure that no method has been started 2 Setalow flow in System Control Manual Pump Flow for example 0 5 ml min 3 Click Execute 4 Set the inlet valve to the appropriate position in System Control Manual Flowpath InletValve for example inlet A11 to A18 or A2 5 Connect a syringe to the purge valve AKTAexplorer AKTApurifier and AKTAmicro Operating Instructions 28 9597 57 AA 33 4 Operation 4 4 Preparations before start 7 Slowly draw solution into t
30. and containers shall be secured and sealed to prevent accidental spillage CAUTION Make sure waste container is dimensioned for maximum possible volume when the instrument is left unattended KTAexplorer KTApurifier and AKTAmicro Operating Instructions 28 9597 57 AA 15 NOTICE Avoid condensation by letting the unit equilibrate to ambient temperature 2 Safety instructions 2 1 Safety precautions Using flammable liquids When using flammable liquids with the AKTAexplorer AKTApurifier and AKTAmicro instrument follow these precautions to avoid any risk of fire or explosion WARNING Fire Hazard Before starting the system make sure that there is no leakage WARNING A fume hood or similar ventilation system shall be installed when flammable or noxious substances are used WARNING Always use appropriate personal protective equipment during operation and maintenance of AKTAexplorer AKTApurifier and AKTAmicro system WARNING Hazardous substances When using hazardous chemical and biological agents take all suitable protective measures such as wearing protective glasses and gloves resistant to the substances used Follow local and or national regulations for safe operation and maintenance of the system AKTAexplorer AKTApurifier and AKTAmicro Operating Instructions 28 9597 57 AA Personal protection 16
31. anufacturer s instructions Wait for the computer to start up Verify that the power indicator on the CU 950 is on when the computer has been turned on Log on to Windows Start UNICORN by double clicking on the UNICORN shortcut icon on the Windows desktop Ab a7 UNICORN In the Logon dialog select a user from the User name list and enter the password If you log on for the very first time select user default and enter the password default Click OK IINE e UNICORN logon User name default Y UNICORN starts and the UNICORN Manager window opens see Figure 4 1 Note See the UNICORN user documentation for instructions about how to create new users 1 AKTAexplorer AKTApurifier and AKTAmicro Operating Instructions 28 9597 57 AA 31 4 Operation 4 3 Starting the control system IE Methods E E Results CA default CA default Size Type Modified Created Name Prev Folder E AC 1 sample ETT 248KB Method File 02 19 2009 09 22 02 20 2009 13 39 Boz162009 5 6 4 samples ETT 261KB Method File 02 24 2009 10 51 02 24 2009 10 51 C3o2172009 iz AC 5 samples ETT 265KB Method File 02 24 2009 13 14 02 24 2009 10 51 03102009 is Immunitettesti ETT 259KB Method File 05 05 2009 17 16 05 06 2009 13 37 03112009 EIT 241KB Method File 05 06 2009 13 47 03 11 2009 17 31 05062009 05122009 No Description 1 The Instant Run icon immediately starts the system control wizard used to start
32. arameter IP20 100 120 220 240 V 50 to 60 Hz 600 VA T6 3 AL 250V 450 x 490 x 610 mm 41kg 4 to 40 C 10 to 95 84 to 106 kPa 840 to 1060 mbar IP20 100 120 220 240 V 50 to 60 Hz 370 VA T 6 3 AL 250 V 450 x 480 x 610 mm 55 kg 4 to 40 C 10 to 95 84 to 106 kPa 840 to 1060 mbar IP20 100 120 220 240 V 50 to 60 Hz 600 VA T6 3 AL 250V 450 x 480 x 610 mm 66 8 kg 4 to 40 C 10 to 95 84 to 106 kPa 840 to 1060 mbar Ingression protec tion Supply Voltage Power consump tion Fuse specification Dimensions H x W x D Weight Ambient tempera ture Relative humidity tolerance non condensing Atmospheric pres sure 7 2 Chemical resistance Chemical Exposure Exposure CAS no Comments gt 1 day up to 2 months Acetaldehyde OK OK Acetic acid lt 5 OK OK AKTAexplorer AKTApurifier and AKTAmicro Operating Instructions 28 9597 57 AA 59 Comments FFKM PP and PE swell PVDF is affected by long term use Silicone is affected by long term use Kalrez CTFE PP and PE are affected by long term use PVDF is affected by long term use ETFE PP PE and PVDF are affected by long term use 200 580 7 200 835 2 231 635 3 235 186 4 231 984 1 249 576 7 200 664 3 200 837 3 64 19 7 75 05 8 7664 41 7 1
33. ate Sodium borate Sodium carbonate Sodium chloride Sodium hydroxide 2M Sodium sulphate Sulphuric acid dilut ed Sulphuric acid medium concentra tion Tetrachloroethy lene Tetrahydrofuran Toluene Trichloroacetic acid 1 Trifluoroacetic acid 1 Urea 8M 62 7 Reference information 7 2 Chemical resistance Chemical Exposure Exposure Comments gt 1 day upto 2 months o Xylene and p Xy OK Avoid PP and PE are af lene fected by long term use 7 3 System recommendations Refer to AKTAexplorer AKTApurifier and AKTAmicro User Manuals or contact your local GE Healthcare representative for the most current information 7 4 Literature For further information regarding the AKTAexplorer AKTApurifier and AKTAmicro system refer to the following e AKTAexplorer AKTApurifier and AKTAmicro User Manuals e AKTAexplorer AKTApurifier and AKTAmicro Safety Handbooks e AKTAexplorer AKTApurifier and AKTAmicro Installation Guides e UNICORN User Manual 7 5 Ordering information For ordering information visit www gelifesciences com AKTA AKTAexplorer AKTApurifier and AKTAmicro Operating Instructions 28 9597 57 AA 63 A Electrical and communication connections Appendix A Electrical and communication connections No Description No Description 1 Fraction collector optional 5 CU 950 2 Ma
34. ation This chapter provides instructions for the use of AKTAexplorer AKTApurifier and AKTAmicro 4 1 Operation overview Workflow The typical workflow in AKTAexplorer AKTApurifier and AKTAmicro after turning on the system and connecting it to UNICORN can be divided into a number of steps Step Action Section Create a method Create a method on page 35 Prepare the system for a run Section 4 4 Preparations before start on page 32 Start a run using a method Section 4 6 Performing a run on page 39 During a run view and change parameters Viewing the run on page 40 Procedures after a run Section 4 7 Procedures after a run on page 42 Evaluate the results See UNICORN user documentation 1 2 3 Liquid flow path See Appendix B Connection diagram Liquid flow path on page 65 for an illustration of the liquid flow path in KTAexplorer AKTApurifier and AKTAmicro 4 2 Starting the instrument Ensure that the waste container and needed buffer bottles are correctly connected Check that all tubing connections are properly tightened and that all valves are connected to a tube or termination Turn on the Power switch on the instrument AKTAexplorer AKTApurifier and AKTAmicro Operating Instructions 28 9597 57 AA 30 4 Operation 4 3 Starting the control system 4 3 Starting the control system Starting UNICORN Turn on the monitor computer and optional printer according to the m
35. covers or replace parts unless specifically stated in the user documentation WARNING Disconnect power Always disconnect power from the instrument before replacing any component on the instrument unless stated otherwise in the user documentation WARNING Hazardous chemicals during maintenance When using hazardous chemicals for system or column cleaning wash the system or columns with a neutral solution in the last phase or step AKTAexplorer AKTApurifier and AKTAmicro Operating Instructions 28 9597 57 AA 43 WARNING Do not perform any type of maintenance work while the system is powered electrically or when the piping system is pressurized Note that the piping system can be pressurized even when the system is closed down NOTICE 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 NOTICE Cleaning Keep the instrument dry and clean Wipe regularly with a soft damp tissue and if necessary a mild cleaning agent Let the instrument dry completely before use 5 Maintenance 5 1 General 5 2 User maintenance schedule Table 5 1 provides a guide to maintenance operations and intervals at which these operations should be performed by the user The user is however responsible for deciding the type
36. dues in the flow path from previ ous runs Air in the eluents Residue in the column from previous runs Incorrect mixer func tion Bad UV fiber connec tions Dirty UV cell Impure buffer Air in the pump or in the UV cell 6 1 UV curve problems Error symptom Ghost peak Noisy UV signal sig nal drift or instability 52 6 Troubleshooting 6 1 UV curve problems Corrective action Check the lamp run time according to and replace if necessary Refer to AKTAexplorer AKTApurifier and AKTAmicro User Manuals Check that the lamp position and the filter position are both set to the wavelength to be used 280 nm or 254 nm Refer to AKTApurifier and AKTAmicro User Manuals Does not apply to AKTAexplorer and AKTApurifier without UPC Calculate the theoretical extinction coefficient of the protein If it is ze ro or very low at 280 nm the pro tein cannot be detected Corrective action Use a flow restrictor after the flow cell Tighten the connectors If neces sary replace the connectors Check the mixer function by plac ing a stirrer bar on top of the mixer housing The stirrer bar should ro tate when the system is in Run mode The mixer function can also be checked by running the installa tion test Clean the conductivity cell by flushing 1M NaOH or 20 ethanol Equilibrate the column If neces sary clean the column Refer to AKTAexplorer AKTApurifier and
37. e V1 14 Outlet Valve V4 7 Restrictor 15 To Fraction Collector optional 8 Sample Valve V5 66 AKTAexplorer AKTApurifier and AKTAmicro Operating Instructions 28 9597 57 AA B Connection diagram Liquid flow path Liquid flow path for AKTApurifier 10 and 100 G7 H7 d LI LI a LI L LI LI LI No Description No Description 1 Pump A 7 Bypass 2 Pump B 8 Column 3 Mixer 9 Flow restrictor 4 On line filter 10 To Fraction Collector optional 5 Injection Valve V1 11 Cond optional Connected after the UV cell 6 Sample loop 12 pH optional Connected after the conductivity cell 67 AKTAexplorer AKTApurifier and AKTAmicro Operating Instructions 28 9597 57 AA B Connection diagram Liquid flow path Liquid flow path for AKTAmicro G3 9 5 UV COND 10 A2 W1 No Description No Description 1 Pump A 6 Sample loop 2 Pump B 7 Bypass 3 Mixer 8 Column 4 On line filter 9 Flow restrictor supplied 5 Injection Valve V1 10 To Fraction Collector optional 68 AKTAexplorer AKTApurifier and AKTAmicro Operating Instructions 28 9597 57 AA 6 Tubing Appendix 6 Tubing Tubing specifications for AKTAexplorer Names in the Label column in Table C 1 and Table C 2 refer to tubing labels in the liquid flow path connection diagram see Liquid flow path for AKTAexplorer 1
38. e 2009 07 08 08 28 23 Central European Standard Time Manual 3 21 ml Continue 2009 07 08 08 28 46 Central European Standard Time Manual 2 00 ml AutozeroU V 1 00 ml Gradient 100 0 B 0 00 ml 0 90 ml Gradient 0 0 B 0 00 ml 0 00 ml InjectionMark 4 00 ml End Block 4 00 ml Block Pure sample 4 00 ml Base SameAsMain 4 00 ml New Chromatogram Absmax 4 00 ml Gradient 100 0 B 0 00 ml For Help press F1 Pun 40 AKTAexplorer AKTApurifier and AKTAmicro Operating Instructions 28 9597 57 AA 4 Operation 4 6 Performing a run Up to four view panes Run Data Curves Flow scheme and Logbook can be displayed showing different aspects of the run in real time The Run Data view pane displays the current values for selected run parameters e The Curves view pane displays the monitor signal values graphically The Flow scheme view pane displays a graphical representation of the chromatography system that shows the current status of the run During a run the flow scheme shows open flow path s in color and monitor signals with numerical displays e The Logbook view pane shows the actions as the run proceeds All actions and unexpected conditons are logged with date time and current user name The log book provides a complete history of the run and is saved in the result file Customize the view panes To customize the view panes right click in the respective view pane and select Proper
39. ected properly Checkthat the electrode is correct ly inserted in the flow cell and if necessary hand tighten the nut Check that the pH electrode is not broken Calibrate the pH electrode Clean the pH electrode if required see Monitor pH C 900 User Manu alor UPC 900 User Manual Compare the response of the pH electrode with that of another pH electrode If the response differ greatly the electrode may require cleaning or replacement In organic solvents such as ethanol methanol and acetonitrile stable pH measurements are not possible since dehydration of the membrane will occur It is recom mended that the pH electrode is not used in applications using or ganic solvents Mount the dummy electrode instead Incorrect pump or Check that the pump and valves valve operation operate correctly Air in the flow cell If air in the flow cell is suspected tap the flow cell carefully or tilt it to remove the air Alternatively flush the cell with buffer at 20 ml min AKTAexplorer 100 and AK TApurifier 100 or 10 ml min AKTA explorer 10 and AKTApurifier 10 or 0 5 ml min AKTAmicro for 1 2 min Use the flow restrictor FR 902 after the pH electrode Static interference AKTAexplorer AKTApurifier and AKTAmicro Operating Instructions 28 9597 57 AA 57 Corrective action There may be interference from static fields Connect the pH flow cell and the rear panel of the monitor using a standard
40. et 49 SealiBRtlonimeebeedemiibereereed idunt 50 5 8 eusanecnauasteariacensaa ersereaaanoureer esate EE 50 b latest uates ior Tireni Eh UY Cave E EE Teni enaiis 52 61 CUPVS 52 6 2 Conductivity curve problems ecsssesssessssssccssssssssssssesssssccssssssssnssssssessssseccssssnsssssessecseccessssssseaeeses 53 6 5 DphHic rve problems snae eret Parere ere be RR i derer efe 55 6 4 Pressure curve problems 58 AKTAexplorer AKTApurifier and AKTAmicro Operating Instructions 28 9597 57 AA 3 Table of Contents 7 Reference 59 A1 ce adam leoi qd ced 59 7 2 Chemical fesIStarice etre ice ter tet b ette re ert eoe Harb d ttende Ioue 59 GS System recommendations 63 74 Literature sss 63 7 5 Ordering information 63 A Electrical and communication connections eere enne 64 B Connection diagram gt Liquid flow path nece rrretrrrerrettn cene o rho reioas 65 TUDNO gt 69 4 AKTAexplorer AKTApurifier and AKTAmicro Operating Instructions 28 9597 57 AA 1 Introduction 1 Introduction Purpose of the Operating Instructions The Operating Instructions provides you with the instructions needed to handle the AKTAexplorer AKTApurifier and AKTAmicro systems in a safe way Prerequisites In order to operate the systems in the way they are intended the following pre requisites must be fulfilled e You should have a general
41. he syringe When fluid starts to enter the syringe continue to draw a few milliliters before closing the purge valve Check that there is no visible air left in the tubing 8 Repeat for the other purge valve 9 To fill inlet B1 and B2 a In System Control Manual Pump Gradient select Target 10096 B and inlet B1 to fill B1 or inlet B2 to fill B2 Wait for the valve to turn a clicking sound before starting the purging procedure b When all inlets are filled click End Connect columns and Superloop For column positions see the method Remove air before connecting columns Air remaining in the system may be removed by purging the pump and by selecting Pump Wash and System Wash 1 Immerse A1 tubing in the buffer to be used 2 Select System Control Manual Pump PumpWash Connecting tubing to columns Refer to column manufacturer s instructions 34 AKTAexplorer AKTApurifier and AKTAmicro Operating Instructions 28 9597 57 AA 4 Operation 4 4 Preparations before start Column attachment drop to drop Attach columns manually by starting a low flow see Purging the pump and inlet tubing on page 33 and selecting System Control Manual Flowpath ColumnPosition Preparing the fraction collector Place the rack chosen in the method in the fraction collector and fill it with appropriate tubes and or deep well plates 4 5 Setting up a run Create a method To create a method 1 Clickthe New Method icon in the UNICO
42. ins supply socket 6 UniNet 1 cable 3 UniNet 1 cable 7 USB cable to computer 4 Power cord 8 Power converter Figure A 1 Electrical and communication connections for AKTAexplorer AKTApurifier and AKTAmicro 64 AKTAexplorer AKTApurifier and AKTAmicro Operating Instructions 28 9597 57 AA B Connection diagram Liquid flow path Appendix B Connection diagram Liquid flow path Liquid flow path for AKTAexplorer 10 A11 A12 A13 A14 A15 A16 A17 A18 No Description No Description 1 Buffer Valve V6 7 Column Selection Valve V2 2 Pump A 8 Column Selection Valve V3 3 Pump B 9 Flow restrictor 4 Mixer 10 Outlet Valve V4 5 On line filter 11 To Fraction Collector optional 6 Injection Valve V1 AKTAexplorer AKTApurifier and AKTAmicro Operating Instructions 28 9597 57 AA 65 8 Connection diagram Liquid flow path Liquid flow path for AKTAexplorer 100 65 63 z I 2 Gi2 1 6 ETA O uS KA 62 COND UV TN 9 T w T 5 7 15 8 UYU 9 60 we w 619 8 ES 14 YIN As NOS Nay UU VYOUVVVVVV U A11 A12 A13 A14 A15 A16 A17 A18 S1 S2 S3 S4 S5 S6 S7 S8 W3 F4 FS F6 F7 FB No Description No Description 1 Buffer Valve V6 9 Sample Pump 2 Pump A 10 Flow Direction Valve V7 3 Pump B 11 Column Selection Valve V2 4 Mixer 12 Restrictor 5 On line filter 13 Column Selection Valve V3 6 Injection Valv
43. ired to lift the system safely NOTICE Disconnect power To prevent equipment damage always disconnect power from the AKTAexplorer AKTApurifier and AKTAmicro instrument before an instrument module is removed or installed or a cable is connected or disconnected NOTICE AKTAexplorer AKTApurifier and AKTAmicro shall be installed and prepared by GE Healthcare personnel AKTAexplorer AKTApurifier and AKTAmicro Operating Instructions 28 9597 57 AA 2 Safety instructions o OP P b 2 1 Safety precautions 18 2 Safety instructions 2 1 Safety precautions 19 WARNING Hazardous chemicals during run When using hazardous chemicals run System CIP and Column CIP to flush the entire system tubing with distilled water before service and maintenance WARNING Hazardous biological agents during run When using hazardous biological agents run System CIP and Column CIP to flush the entire system tubing with bacteriostatic solution e g NaOH followed by a neutral buffer and finally distilled water before service and maintenance CAUTION Hazardous chemicals in UV flow cell Make sure that the entire flow cell has been flushed thoroughly with bacteriostatic solution for example NaOH and distilled water before service and maintenance NOTICE Do not run Column CIP if using silica based packing material or RPC columns Remove the column from the system during CIP WARNING
44. ked AKTAexplorer AKTApurifier and AKTAmicro Operating Instructions 28 9597 57 AA AKTAexplorer AKTApurifier and AKTAmicro are high pressure liquid chromatography systems for use in laboratory scale production of biomolecules Figure 1 1 The main parts of the instruments AKTAexplorer is shown as an example Part Function Monitor pH C 900 Monitor UV 900 Pump P 900 P 901 alt P 903 Power switch Switch valve SV 903 Conductivity cell UV cell AKTAexplorer AKTApurifier and AKTAmicro Operating Instructions 28 9597 57 AA 1 Introduction 1 3 Instrument 1 5 Instrument Product description 1 2 3 10 1 Introduction 1 5 Instrument Part Function 8 Mixer M 925 9 Sample pump P 960 This is only standard in some versions 10 Valve door 11 Column selection valve V3 PV 908 12 Sample valve V5 PV 908 13 Injection valve V1 INV 907 14 Column selection valve V2 PV 908 15 Flow direction valve V7 INV 907 This is only standard for AKTAExplorer 100 and AKTAExplorer 100 Air 16 Outlet valve V4 PV 908 17 Buffer valve V6 INV 908 18 Box 900 Detailed information on the components included in each system can be found in their respective User Manual Basic flow path T T ii 5 WAY W Ln
45. laborato ry 4mm banana plug cable Replace the pH electrode Corrective action Check all connections for leaks Check that there is sufficient eluent present in the reservoirs Use degassed solutions Purge the pump Follow the instructions in Pump P 900 User Manual Clean the valves in according to Pump P 900 User Manual Replace the piston seal according to the instructions in Pump P 900 User Manual Flush through to clear blockage If necessary replace tubing Check inlet tubing filter It can be come clogged if unfiltered buffers or samples are applied See instruc tions for flushing through at the end of the run in Pump P 900 User Manual AKTAexplorer AKTApurifier and AKTAmicro Operating Instructions 28 9597 57 AA Possible cause Problem with the electrode 6 4 Pressure curve problems Possible cause Air bubbles passing through or trapped in the pump Inlet or outlet check valves not functioning correctly Piston seal leaking Blockage or part blockage of flowpath 6 Troubleshooting 6 3 pH curve problems Error symptom pH values vary with varied back pressure Error symptom Erratic flow noisy baseline signal irregu lar pressure trace 7 Reference information 7 Reference information This chapter contains technical data regulatory and other information AKTApurifier AKTAmicro AKTAexplorer 7 1 Specifications P
46. ls on liquid flow path see Appendix B Connection diagram Liquid flow path on page 65 UNICORN is a complete software for control and supervision of AKTAexplorer AKTApurifier and AKTAmicro The software runs under Microsoft Windows operating system UNICORN is supplied with a method wizard which provides easy creation of methods for purification AKTAexplorer AKTApurifier and AKTAmicro Operating Instructions 28 9597 57 AA 1 Introduction 1 3 Instrument 1 4 Control software UNICORN control software 12 1 Introduction 1 4 Control software For more information about UNICORN control system see the UNICORN user manuals supplied AKTAexplorer AKTApurifier and AKTAmicro Operating Instructions 28 9597 57 AA 13 2 Safety instructions 2 Safety instructions This chapter describes safety compliance safety labels general safety precautions emergency procedures power failure and recycling of AKTAexplorer AKTApurifier and AKTAmicro 2 1 Safety precautions Introduction The AKTAexplorer AKTApurifier and AKTAmicro instruments are powered by mains voltage and handles pressurized liquids that may be hazardous Before installing operating or maintaining the system you must be aware of the hazards described in this manual Follow the instructions provided to avoid personal injury or damage to the equipment The safety precautions in this section are grouped into the following categories General precautions
47. n Section 4 6 Performing a run on page 39 Cleaning columns When running different types of purification methods and different samples after each other the columns should be cleaned between the runs according to the column instructions This will remove unspecific bound proteins and prevent column clogging 1 Inthe Method Editor module in UNICORN create a method for column cleaning in place CIP 2 Immerse the tubing ends to be used in the correct containers according to the method for the chosen run 3 Run the cleaning in place method as described in Section 4 6 Performing a run on page 39 42 AKTAexplorer AKTApurifier and AKTAmicro Operating Instructions 28 9597 57 AA 5 Maintenance 5 Maintenance This chapter provides instructions for routine component maintenance and a maintenance schedule 5 1 General Regular maintenance is important for safe and trouble free operation of your instrument The user should perform daily and monthly maintenance Preventive maintenance should be performed on a yearly basis by qualified service personnel For maintenance of a specific component carefully read the component manual and follow the instructions To avoid personal injury when performing maintenance on the AKTAexplorer AKTApurifier and AKTAmicro instrument follow the instructions below WARNING Electrical shock hazard All repairs should be done by service personnel authorized by GE Healthcare Do not open any
48. nation at work
49. on 1800 9 11 9 x 103 Switch valve pump A3 B3 Teflon 150 2 9 991 Pump Mixer G1 G2 PEEK 300 0 75 133 Mixer On line filter G3 PEEK 150 0 75 66 Filter V1 Inj Valve G4 PEEK 460 0 75 203 V1 V7 G5 PEEK 470 0 75 208 V7 V2 G6 PEEK 410 0 75 181 Bypass G7 PEEK 620 0 75 274 V3 V7 G8 PEEK 470 0 75 208 V7 loop G9 PEEK 180 0 75 80 V7 UV G10 PEEK 370 0 75 163 UV Cond Cell G11 Tefzel 160 0 75 71 Cond Cell pH Cell G12 Tefzel 450 0 75 199 pH Cell Flow restrictor G13 Tefzel 110 0 75 49 Flow restrictor 1 G14 Tefzel 120 0 75 53 V4 Fraction collector G15 Tefzel 600 0 75 265 Flow through F3 Tefzel 1000 1 0 785 V1 Waste W1 Tefzel 1300 0 75 574 70 KTAexplorer AKTApurifier and AKTAmicro Operating Instructions 28 9597 57 AA 6 Tubing Material Length Volume mm mm ul Sample pump Waste w2 Teflon 1300 1 6 2 6 x 10 V4 Waste W3 Tefzel 1000 1 0 785 1 Not mounted at factory Tubing specifications for AKTApurifier Names in the Label column in Table C 3 and Table C 4 refer to tubing labels in the liquid flow path connection diagram see Liquid flow path for AKTApurifier 10 and 100 on page 67 Table C 3 Tubing specifications for AKTApurifier 10 Material Volume ul Inlet A1 Al Teflon 1700 16 3 4 x 10 Inlet A2 A2 Teflon 1900 16 3 8 x 103 Inlet B1 B1 Teflon 1500 16 3 0 x 10 Inlet B2
50. or measurement control CAN CSA C22 2 and laboratory use No 61010 1 8 KTAexplorer AKTApurifier and AKTAmicro Operating Instructions 28 9597 57 AA 1 Introduction 1 2 Regulatory information Notes Harmonized with 2004 108 EC Harmonized with 2006 42 EC Harmonized with 2006 42 EC Description EMC emissions and immunity require ments for electrical equipment for measurement control and laboratory use Safety of machinery Basic concepts general principles for design Safety of machinery Principles of risk assessment Standard EN 61326 1 EN ISO 12100 1 12100 2 EN ISO 14121 1 14121 2 The CE marking and the corresponding Declaration of conformity is valid for the instrument when it is e used as a stand alone unit or connected to other CE marked instruments or e connected to other products recommended or described in the user documentation and e usedinthe same state as it was delivered from GE Healthcare except for alterations described in the user documentation The Declaration of conformity is valid only for systems that are marked with the CE marking CE CE Marking Regulatory compliance of connected equipment Any equipment connected to the AKTAexplorer AKTApurifier and AKTAmicro systems should meet the safety requirements of EN 61010 1 IEC 61010 1 or relevant harmonized standards Within EU connected equipment must be CE mar
51. r 2 connect a termination plug to the empty UniNet 1 socket Frac 950 only Flow path All parts and tubing are mounted on the AKTAexplorer and AKTAmicro systems at delivery AKTApurifier has no preconnected tubing It is recommended that the mounting is done by GE Healthcare service engineers For more information regarding installation see AKTApurifier User Manual Setting the delay volume in UNICORN The delay volume is volume of liquid in the flow path from the UV sensor that identifies the peakto the fraction collector The length of tubes affects the delay volume that needs to be changed in UNICORN To change the delay volume in UNICORN 1 Select System Settings in System Control 2 Select Specials and then FracParameters 3 Enterthe delay volume and click OK Note To prevent bacterial growth the system flow path is filled with 2096 ethanol at delivery e Connect tubes for reagents solvents and sample collection to the correct inlet and outlet connections on the system For more information see AKTAexplorer AKTApurifier and AKTAmicro User Manuals Electrical power Connect the power cord to a grounded power outlet specified in Section 5 1 Site requirements on page 26 5 6 Spare parts and accessories For correct up to date information on spare parts and accessories visit www gelifesciences com AKTA AKTAexplorer AKTApurifier and AKTAmicro Operating Instructions 28 9597 57 AA 29 4 Operation 4 Oper
52. r Insufficient mixing chamber volume Incorrect motor func tion Air in the flow cell Dirty tubing Incorrect mixer vol ume 6 Troubleshooting 6 2 Conductivity curve problems Error symptom Conductivity measure ment with the same buffer appears to de crease over time Waves on the gradi ent Ghost peaks appear in the gradient profile Unlinear gradients or slow response to B changes 54 6 Troubleshooting 6 2 Conductivity curve problems Corrective action Check that the conductivity flow cell cable is connected properly Check that the pump and valves operate correctly If temperature compensation is being used check that the temper ature sensor is calibrated and that the correct temperature compen sation factor is in use Check that the column is equilibrat ed If necessary clean the column Check the operation of the mixer The mixer function is checked by placing a stirrer bar on top of the mixer housing The stirrer bar should rotate when the system is in Run mode The mixer function can also be checked by running the installation test Corrective action Check that the electrode cable is connected properly The electrode glass membrane may be cracked Replace the elec trode Check that the pH monitor is cor rectly connected according to the User Manual Clean the pH electrode as detailed in Monitor pH C 900 User Manual or UPC 900
53. rol unit between unit and computer e Various buffer and sample bottles Installing the pH electrode Install the pH electrode in the flow cell according to the picture below AKTAexplorer AKTApurifier and AKTAmicro Operating Instructions 28 9597 57 AA 27 3 Installation 3 4 Assembly Description 5 Description 1 To instrument rear pH probe 5 Flow cell holder 2 Electrode 6 Flow cell 3 Nut 7 End cover 4 Dummy The flow cell holder can be placed on either the optical unit for AKTApurifier or on the outside of the valve door for AKTAexplorer For more information on installation see AKTApurifier User Manual and AKTAexplorer Installation Guide 3 5 Connections Communication Connect the network signal cables and computer according to the electrical drawings in Appendix A Electrical and communication connections on page 64 Ensure that UNICORN control software is installed on the computer Installing Controller unit CU 950 Hang the CU 950 on the left side of the system by inserting the hooks on the front of CU 950 into the channel on the side of the UV 900 28 AKTAexplorer AKTApurifier and AKTAmicro Operating Instructions 28 9597 57 AA 3 Installation 3 5 Connections Connect according to diagram in Appendix A Electrical and communication connections on page 64 When using a fraction collector 1 connecta UniNet 1 cable between Monitor UPC 900 Monitor UV 900 and the fraction collecto
54. s contain hazardous substances Detailed information is available from your GE Healthcare representative 24 AKTAexplorer AKTApurifier and AKTAmicro Operating Instructions 28 9597 57 AA 2 Safety instructions 2 4 Recycling information Disposal of electrical components Waste of electrical and electronic equipment must not be disposed as unsorted municipal waste and must be collected separately Please contact an authorized representative of the manufacturer for information concerning the decommissioning of equipment AKTAexplorer AKTApurifier and AKTAmicro Operating Instructions 28 9597 57 AA 25 3 Installation 3 Installation NOTICE 9 AKTAexplorer AKTApurifier and AKTAmicro shall be installed and prepared by GE Healthcare personnel AKTAexplorer AKTApurifier and AKTAmicro are delivered in protective packing material and shall be unpacked with great care Any equipment connected to AKTAexplorer AKTApurifier and AKTAmicro must fulfill applicable standards and local regulations For detailed information on Installation see AKTAexplorer AKTApurifier and AKTAmicro Installation Guides 3 1 Site requirements Parameter Requirement Electrical power 100 240 V 50 60 Hz Ambient temperature 4 C to 40 C Placement Stable laboratory bench min 200 x 80 cm Humidity 20 to 9596 non condensing 3 2 Transport The equipment weights are specified in the table below Each system requires at least
55. ties For more information about customizing the view panes see the UNICORN user documentation Ending the run To stop the run on a system before it is finished Click End above the Run data view pane Status indicator colors The status indicator is located at the bottom of System Control The table below shows how the indicator colors relate to the run status Indicator color Run status White End Green Run or Manual Yellow Hold Red Pause KTAexplorer KTApurifier and AKTAmicro Operating Instructions 28 9597 57 AA 41 4 Operation 4 6 Performing a run Error indication When a warning or an alarm is issued from a system an error code is displayed See AKTAexplorer AKTApurifier and AKTAmicro User Manuals for guidance Evaluate the results See AKTAexplorer AKTApurifier and AKTAmicro User Manuals and UNICORN user documentation for how to evaluate the results 4 7 Procedures after a run Cleaning system To keep the system in good shape it is important to clean both the tubing and the outside of the system regularly 1 Inthe Method Editor module in UNICORN create a method for cleaning the system 2 Wash the outside of the inlet tubings with water and or ethanol 3 Immerse the tubing ends to be used in the container with cleaning solution 4 If the column valve is to be cleaned remove the columns and reconnect the tubings to the column valves 5 Run the cleaning method as described i
56. tion valve manually to Waste position Put the open end in a waste container 3 Run the pump manually at 10 ml min with water Note the back pressure Bp1 on the pump display or in the Run Data window 4 Setthe system to Pause and connect the flow restrictor to the open end of the capil lary lobserve the IN marking Put the flow restrictor in the waste container 5 Press Continue so that the pump run at 10 ml min with water Note the back pressure Bp2 on the pump display or in the Run Data window 6 Calculate the back pressure generated by the flow restrictor Bp2 Bp1 Replace it if it is not within limit Yearly Valve inspection Check for external or internal leakage Replace channel plate and distribution plate yearly or when required Refer to the relevant valve in struction sheet 5 5 Cleaning Cleaning before maintenance service Before maintenance service is performed the system owner must first clean the system and complete a Decontamination Report Contact GE Healthcare for further information AKTAexplorer AKTApurifier and AKTAmicro Operating Instructions 28 9597 57 AA 47 5 Maintenance 5 3 Cleaning Cleaning In Place All components in the system are designed for ease of CIP After repeated separation cycles contaminating material might progressively build up in the system and on the column This material may not have been removed by the cleaning step described above The nature and
57. understanding of how the PC and Windows works e You should understand the concepts of liquid chromatography e You must read and understand the Safety Instructions e The instrument and software should be installed configured and calibrated according to the Installation Guide In this chapter This chapter contains important user information and a general description of the AKTAexplorer AKTApurifier and AKTAmicro systems and their intended use AKTAexplorer AKTApurifier and AKTAmicro Operating Instructions 28 9597 57 AA 5 1 Introduction 1 1 Important user information 1 1 Important user information Read this before using the AKTAexplorer AKTApurifier and AKTAmicro system G All users must read the safety instructions in the KTAexplorer KTApurifier and AKTAmicro User Documentation to fully understand the safe use of the systems before installing using or maintaining the systems Do not operate the AKTAexplorer AKTApurifier and AKTAmicro systems in any other way than described in the user documentation Otherwise you may be exposed to hazards that can lead to personal injury and you may cause damage to the equipment Intended use AKTAexplorer AKTApurifier and AKTAmicro are liquid chromatography systems intended for protein purification within method development and drug discovery The systems can be used to screen for optimal choice of columns media and running parameters to purify selected proteins
58. ur local GE Healthcare representative AKTAexplorer AKTApurifier and AKTAmicro Operating Instructions 28 9597 57 AA 49 5 Maintenance 5 6 Replacement of fuses WARNING For continued protection from fire hazard replace only with same type and rating of fuse 5 7 Calibration The table below lists the type and frequency of calibrations that can be done on the instrument Refer to UNICORN user documentation and to the individual component User Manuals and Instructions for descriptions of how to perform these calibrations The calibrations are performed from UNICORN by selecting System Calibrate in System Control Component How often pH monitor if applicable Every day Pump if applicable Whenever the running conditions are changed e g viscosity of sample or buffer temperature backpressure etc If the sample pump is not used frequently it should be calibrated before use Pressure reading When required Conductivity Cell constant Only necessary if specific conductivity with high flow cell accuracy is measured Cond Calib Temperature Must be done when changing the conductivity flow cell Temp Entering a Must be done when changing the conductivity new cellcon flow cell Cond Cell stant 5 8 Storage General recommendation For storage the system must first be cleaned as described in Cleaning In Place on page 48 After cleaning the system must be filled with 0 01

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