Home

Getting Started with ÄKTA avant

1. 8 When buffer leaves the tubing on port 2A in a continuous mode and the top part of the column is filled with buffer connect the tubing to the top of the column 9 Connect a piece of tubing to the bottom of the column 10 When buffer leaves the tubing at the bottom of the column in a continuous mode connect this piece of tubing to the Column valve Use the port opposite to the one already connected to the column in this example port 2B 92 Getting Started with AKTA avant 29 0351 83 AA 6 Prepare the system for a run 6 4 Connect a column Step Action 11 Click the End icon in the System Control toolbar to end the run man Getting Started with KTA avant 29 0351 83 AA 93 6 Prepare the system for a run 6 5 Calibrate the pH monitor 6 5 Calibrate the pH monitor Introduction If pH will be measured during the chromatographic run the pH monitor should be cali brated before the run is started Use two pH calibration buffers with a difference of at least one pH unit Preferably use a pH standard buffer pH 4 or 7 as the first calibration point and a pH standard buffer close to the lowest or highest pH you need to measure as your second point Allow the buffers to reach the operating temperature before use Note Do not run a system flow during pH calibration Calibrate the pH monitor Follow the instruction below to perform the calibration Step Action 1 Open the System Control module and select Sy
2. The column is filled with buffer solution Select inlet positions Enter the solution identity volume flow rate and incubation time By adding steps several preparation solutions can be used Getting Started with AKTA avant 29 0351 83 AA 65 5 Create a method 5 1 Guide to method creation 5 1 2 Predefined methods Method Description Desalting DS After equilibration and sample application the proteins are eluted isocratically This technique is commonly used for buffer exchange Gel Filtration GF After equilibration and sample application proteins separate and elute according to their size largest first Hydrophobic In teraction Chro matography HIC After equilibration and sample application use a buffer contain ing a high salt concentration for example 2 M Ammonium Sul phate hydrophobic proteins are adsorbed to the column ligand After a wash to remove unbound sample elution is performed using a gradient of decreasing salt concentration Finally the column is washed and re equilibrated with start buffer Intelligent Pack ing Packs AxiChrom columns with a predetermined column type by a flow of hydraulic liquid that pushes the adaptor down The user initiates the start of compression at the exact point when the adapter reaches the consolidated bed surface The adapter compresses the bed according to the packing factor or target bed height as selected Two Column Performance Test
3. The CE marking and the corresponding EC Declaration of Conformity is valid for the in strument when it is e used as a stand alone unit or 10 Getting Started with AKTA avant 29 0351 83 AA e connected to other GE Healthcare instruments or 1 Introductio n 1 2 Regulatory information 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 International standards This product fulfills the requirements of the following standards Standard EN ISO 12100 Description Safety of machinery General princi ples for design Risk assessment and risk reduction Notes EN ISO standard is har monized with EU direc tive 2006 42 EC EN 61010 1 IEC 61010 1 UL 61010 1 CAN CSA C22 2 No 61010 1 Safety requirements for electrical equipment for measurement con trol and laboratory use EN standard is harmo nized with EU directive 2006 95 EC EN 61326 1 IEC 61326 1 Emission accord ing to CISPR 11 Group 1 class A Electrical equipment for measure ment control and laboratory use EMC requirements EN standard is harmo nized with EU directive 2004 108 EC EN 301 489 1 EN Electromagnetic compatibility and EN standard is harmo Radio spectrum Matters ERM Short Range Devices SRD Radio equip ment in the freq
4. e g UV1_280 for proteins e Select a target peak table for storage of the result e Select Calculate baseline from the Baseline list e Click OK Chromatogram Target peak table Chrom 1 V Chrom 1 Manual Run 001 Result reca 002 UY 2_0_Chrom 1 Manual Run 001 Result recove 003 UY 3_0_Chrom 1 Manual Run 001 Result recove 004 Cond_Chrom 1 Manual Run 001 Result recovere 005 Cond_Chrom 1 Manual Run 001 Result recove _ 006 Conc B_Chrom 1 Manual Run 001 Result recove 007 pH_Chrom 1 Manual Run 001 Result recovered 008 System flow_Chrom 1 Manual Run 001 Result re 009 System pressure_Chrom 1 Manual Run 001 Rest 010 Cond temp_Chrom 1 Manual Run 001 Result rec 013 Sample flow_Chrom 1 Manual Run 001 Result re 014 Sample pressure_Chrom 1 Manual Run 001 Rest 015 PreC pressure_Chrom 1 Manual Run 001 Result 016 DeltaC pressure_Chrom 1 Manual Run 001 Resu z DN Pat arassa Tina Maral Bri D oe ae Peak table name lt ji J gt UV 1_280 01 PEAK 7 Baseline Calculate baseline Peak Window negative pe Baseline Settings Reject Peaks C Peak skim j 10 06 ratio Column height bed height 0 00 em Column Vt ooj Result The Peak Table is displayed beneath the active chromatogram and the start and end points of each peak are marked by vertical marks in the chromatogram Getting Started with AKTA avant 29 0351 83 AA 131 8 Evaluate and print the results 8
5. increasing salt concentration of e g NaCl Finally the column is washed and re equilibrated with start buffer Cation Exchange After equilibration and sample application positively charged Chromatography proteins are adsorbed to the column ligand After a wash to re CIEX move unbound sample elution is performed using a gradient of increasing salt concentration of e g NaCl Finally the column is washed and re equilibrated with start buffer Chromatofocus After equilibration and sample application elution is performed ing CF using a pH gradient The proteins separate and elute according to their isoelectric points Finally the column is re equilibrated Only available for AKTA avant 25 Column CIP The column is filled with a cleaning solution Select inlet positions Enter the solution identity volume flow rate and incubation time By adding steps several cleaning solutions can be used Sugges tions for cleaning steps are available for a number of column types Column Perfor mance Test After equilibration of the column sample is injected and eluted isocratically A non adsorbing sample like acetone or salt should be used After the run calculate column performance in the Evaluation module The efficiency of the column is determined in terms of height equivalent to a theoretical plate HETP and the peak asymmetry factor A The result is logged in the column logbook Column Prepara tion
6. mon parameters used in the subsequent phases Equilibration Equilibrates the column before purification or re equilibrates the column after purification Sample Applica Applies sample to the column Defines the sample application tion technique the sample volume and the handling of flowthrough Column Wash Washes out unbound sample after sample application or re moves strongly bound proteins after elution Elution Elutes the sample from the column Defines parameters for the elution and fractionation settings Column Prepara tion Prepares the column before use by removing the storage solu tion and equilibrating the column By adding steps several preparation solutions can be used sequentially Column CIP Cleans the column after purification runs by rinsing the column with a cleaning solution to remove unspecifically bound pro teins By adding steps several cleaning solutions can be used sequentially System Prepara tion Prepares the system before a run by removing storage solution and filling the system and inlets with buffer solution One preparation solution is used per phase System CIP Cleans the system after purification runs by rinsing the system with a cleaning solution One cleaning solution is used per phase Column Perfor Tests the efficiency of a packed column in terms of height Getting Started with AKTA avant 29 0351 83 AA mance Test equivalent to a theo
7. 9 Maintenance 9 Maintenance Introduction This chapter provides schedules for preventive maintenance that should be performed by the user of AKTA avant Regular maintenance is essential for reliable function and results Refer to the AKTA avant User Manual for detailed instructions WARNING Always use appropriate personal protective equipment during op eration and maintenance of AKTA avant system Maintenance program An overview of the preventive maintenance to be performed on AKTA avant is outlined below See the AKTA avant User Manual for detailed information about the maintenance procedures Maintenance is divided into Daily maintenance Weekly maintenance Monthly maintenance Bi annual maintenance Maintenance when required WARNING 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 documenta tion 140 Getting Started with AKTA avant 29 0351 83 AA 9 Maintenance Periodic maintenance program The following periodic maintenance should be performed by the user of AKTA avant Interval Maintenance action Daily Calibrate the pH monitor Weekly Calibrate pressure monitors Weekly Change pump rinsing solution Weekly Replace online filter in the Mixer Monthly Check the Flow restrictor Bi annual Clean the UV flow cell Bi annual Replace pH el
8. If you want to then 46 access any of the manu als in PDF format select Help Help for in any of the UNICORN modules see illustration above in the TOC pane expand the heading UNICORN 6 online documentation portal and select Documenta tion overview in the PDF manuals section click one of the text links click the illustration or the text link of the manual of interest find information about a method instruction In the Method Editor module open a method select the instruction of interest in the Instruction box in the Text instruction pane press the F1 key In the System Control module select Manual Execute Manual Instructions expand a heading and select the instruction of interest press the F1 key or click the Help icon in the dialog Getting Started with AKTA avant 29 0351 83 AA 4 The instrument and software 4 3 UNICORN software modules 4 3 UNICORN software modules About this section Three of the four UNICORN modules are used in this Getting Started guide and described in this section Method Editor System Control and Evaluation The Administration module and its icons are described in UNICORN 6 Administration and Technical Manual In this section This section contains the following subsections Section See page 4 3 1 Method Editor module 48 4 3 2 System Control module 51 4 3 3 Evaluation module 53 Getting Started with AKTA avant 29 0351 83 AA 47 4
9. Lees gadert Pissed Pocesung Comgseted 220 mi ave Someries 222 mi Graders 100 00 RB 10 00 ICV prag Phocesung 2 29 mi Grader 10000 80 10 00 CVI Kongited Getting Started with AKTA avant 29 0351 83 AA 51 4 The instrument and software 4 3 UNICORN software modules 4 3 2 System Control module System Control toolbar icons The table below shows the System Control toolbar icons that are referred to in this Getting Started Icon Function Open Method Navigator Opens the Method Naviga tor where available meth ods are listed Icon Function Run Starts a method run Hold Suspends the method run while current flow rate and valve positions are sustained ta u Pause Suspends the method run and stops all pumps Continue Resumes for ex ample a held or paused End Permanently ends the method run and run log contents can be set I gt Lal method run l aaa Customize Opens the Cus Connect to Systems Opens A 3 tomize dialog where curve P the Connect to Systems di A f settings run data groups KA alog where systems can be connected and currently connected users are dis played 52 Getting Started with KTA avant 29 0351 83 AA 4 The instrument and software 4 3 UNICORN software modules 4 3 3 Evaluation module 4 3 3 Evaluation module Introduction The Evaluation module is used to evaluate the results from chromato
10. On Oe re renner nioe nro ene eae NDE e AL Niena 55 SL GUI LOMELAOGCLEGEIOM sss sssancsesocsssnsectcssessvosssessseacescssssssvis5igsosaCasseadavasisostobtorsonaunssseinniscctstbendbovsvns 56 5 1 1 Create GAA edit Methods csssossssssssit enstansssssssssiedsesonssdstekiosasessinsnoisessonsssesieonssnasiadsadanssssdsatuisessessssses 57 BTS Predefined Method S ssscestesiscsisccccsssssectssstecssssssecccscsstnensessseuvay asbecscosseaassslaatuasnccsasscetsetasdtastonventsatshs 65 SKS Predefined DR ASOS sssssssccssssscsssssccccsssecsssattvaassbsccssdontesassevayabeehosssonssiesti cass aiin 67 52 PRUE CPM OG asses ocascsszesacceaccussycceoiscvasavaevaazasssceccobecsssesssiustidiastobsosossvan NN NRANG 68 Prepare the system for OTUN ica oo epic 70 6 1 Before you prepare the SyStem ou esssseesssssssssssesssstesssssssstsssssssssmssessssssssseeeecseececceeecceceeceeseeeeeees 71 62 SPREE EMS TOW CELA csscesestassarsstaezscscscisceaasaasssibencdanssdtavangaialicscsoveacevon AAN 73 6 3 Prime buffer inlets and purge system PUMPS Q eeessessessssssssssssssnessssnnsssseseneseesseeseceeeeeeeeees 81 64 Connecta Coles a ana nE INE 89 65 LCA HATS the PH MONILO f csss ssssoccedsasssesvssschussszSoosooshebbediaeseotorssnconsonsesasiuvssoocdeonstaeabisestannlheateassbitsies 94 6 6 Prepare the Fraction collector 96 6 7 Prepare for a run in a cold cabinet 102 RUMME NO RennebnPee Renee tre iene Ree POTne Suen re fcne Pes Bepee een Csers Pe ene DNC Prec nent mr
11. The instrument and software 4 3 UNICORN software modules 4 3 1 Method Editor module 4 3 1 Method Editor module Introduction In UNICORN software the instructions to control a chromatography run are defined in a method The Method Editor module is used to create or edit such methods The graphical user interface combines default settings with the ability to change settings This simplifies method creation and replaces the method wizard in earlier versions of UNICORN Predefined methods and phases A method is built up by a number of phases Each phase represents a major process step in the method for example equilibration or elution Predefined methods that include all the phases necessary to run the system are available for different chromatography techniques and also for system cleaning Refer to UNICORN 6 Method Manual for infor mation about predefined methods and phases and how to create a method 48 Getting Started with AKTA avant 29 0351 83 AA 4 The instrument and software 4 3 UNICORN software modules 4 3 1 Method Editor module Method Editor panes As illustrated below four panes show in the Method Editor by default Available phases are found in the Phase Library 1 and an overview of the phases included in the active method is displayed in a Method Outline 2 Detailed information about the method is presented in the upper right hand pane 3 containing the two tabs Phase Properties and Text Instructions The gr
12. button and select Reset Zoom Result The original area is restored in one step 128 Getting Started with KTA avant 29 0351 83 AA View documentation To view documentation of the run e Click the View Documentation icon to open the Documentation dialog 8 Evaluate and print the results 8 1 View the results e Select the tab of interest Which tabs are displayed depends on the settings in the active method e Click OK to close the dialog Documentation Fraction Collector Variable List Scouting Text Instructions formation Result Notes BuffePro Columns Evaluation Procedures _ i Run Log 0 00 min Batch ID sdfgfihertniztatnz 0 00 min Base Volume 20 106 ml GSTPrep FF 16 10 0 00 min Base SameAsMain 0 02 min Wavelength 280 nm Off nm Off nm Completed 0 02 min Noise reduction UV 5 0 sec Issued Processing NN min Naise reduction II 5 N feer Comnleted 0 00 min Method Run 2009 01 23 09 55 33 01 00 Method Dokumentation Result Moa Dokumentation Dokumentation 001 User Ma 0 00 min The method flow is now calculated based on the system flow System 0 00 min Phase Method Settings Issued Processing Completed 0 00 min Alarm pre column pressure Enabled 20 00 MPa 0 00 MPa Issued Processing 0 01 min Alarm pre column pressure Enabled 20 00 MPa 0 00 MPa Completed 0 01 min Alarm delta column pressure Enabled 3 50 MPa 0 00 MPa Issued Processing 0 01 min A
13. by User AGEL 02759 Getting Started with KTA avant 29 0351 83 AA 7 Runa method 7 4 Monitor the run Part Description 1 Run Data Presents current run data values 2 Chromatogram Illustrates data as curves 3 Run Log Presents all registered actions 4 Flow Scheme Illustrates the current flow path Customize Run Data The Run Data pane shows real time data from available monitors during the run To change what parameters to display in the pane Click the Customize icon to open the Customize dialog Under the Run Data Groups tab se lect the parameters to display Click OK to close the dialog Customize System3 Run Data Groups Curves Curve Style W Rundata System state M Acc volume Block volume M Acc time M Block time Scouting no O System flow C System pressure O Cone B O Sample flow C Sample pressure O Prel pressure O DeltaC pressure O PostC pressure oui Ouve Ouv3 O pH O ButferPro pH O Cone 01 O Cone G2 O Cone 03 O Conc 04 O Cond O Cond Customize Chromatogram The Chromatogram pane displays a graphical presentation of registered data from available monitors during the run To change the chromatogram properties Click the Customize icon to open the Customize dialog Make appropriate selections under the tabs described in the table below Getting Started with KTA avant 29
14. erens 104 Fall Before OU SUC t neninm i ana a6 a aa aaa Naa 105 7 2 Applying the sample au ccesssssessesssssssssssssssssssssesseccssssssssssssssessecesssssssussssssssecessssssssussuscsssscessssessssassees 108 Ta Soramethod BUI amrni a aa tarri Nia nA Ea 111 TA Momtor thE TU suninman RA NARS 116 7 5 After run procedures u esssssssssessessssssssssssssssssssssescccssssssssussussseseccssssssssussuscseseccsassssssuasusssesseessssesssnssnees 121 Getting Started with KTA avant 29 0351 83 AA 3 Table of Contents 8 Evaluate and print the results icscsssscccssicses s dcccssscdvaswviccssecsasseshsdeesdiuveastsacesasbinesss 125 81 View the PESTS saesaassassssccssczsazass hstssnatsssensesddesdsaantanssediastntvon aa ianiai 126 g2 PRK TIMES CHEIOIN sissien u a AARAA sas esbsseteeians ive 130 83 Prntthe FESUIIES a sssss sass lesson dessussinsssicastesshvssvedas cede stetiebsveseivoverbavasbtsddbvadossscebbsscessesesasongabibsasbeedsiaiieh 135 9 UGUIAEOING INGE cccrcnmenni i n n E R E 140 4 Getting Started with KTA avant 29 0351 83 AA 1 Introduction 1 Introduction About this chapter This chapter contains important user information description of safety notices regula tory information and a general description of the intended use of AKTA avant In this chapter This chapter contains the following sections Section See page 1 1 Important user information 6 1 2 Regulatory information 10 Getting Started with AKTA avan
15. in the vicinity of the instrument The facility should comply with the national code of practice for biosafety e High pressure The KTA avant instrument operates under high pressure Wear protective glasses at all times CAUTION To avoid hazardous situations when working with the AKTA avant system take the following measures for personal protection e Always use appropriate personal protective equipment when decommissioning the equipment e Close doors To minimize the risk of exposure to hazardous chemicals and pressurized liquids always close the Foldable door and the Pump cover before starting a run e Cut injuries The tubing cutter is very sharp and must be han dled with care to avoid injuries 18 Getting Started with AKTA avant 29 0351 83 AA Installing and moving the instrument 2 Safety instructions 2 1 Safety precautions WARNING To avoid personal injury when installing or moving the AKTA avant instrument follow the instructions below Heavy object The AKTA avant instrument weighs about 116 kg Use proper lifting equipment or use four or more persons when moving the instrument All lifting and moving must be performed in accordance with local regulations Moving the instrument horizontally Three persons are recom mended when moving the instrument horizontally Supply voltage Make sure that the supply voltage at the wall outlet corresponds to the marking on the instrument before
16. location Undo Restores the method to the state it was in before the last change Redo Restores the method to the state it was in before the Undo command was used 50 Getting Started with KTA avant 29 0351 83 AA 4 The instrument and software 4 3 UNICORN software modules 4 3 2 System Control module 4 3 2 System Control module Introduction The System Control module is used to start view and control a method run System Control panes As illustrated below four panes show in the System Control by default The Run Data pane 1 presents current data in numerical values while the Chromatogram pane 2 illustrates data as curves during the entire method run Information about the method progression is presented in the Run Log 3 and the current flow path is illustrated in the Flow Scheme 4 i ES Se ee how 0157 way TETA hpa jrm o u 8883 S M e 08 66 9 18 Ne TKS ah E ae Rum Log Fater en 220 mi System thr 1 00 indeed Pre cokan permer Pouedi Plocemne 221 wi System Now 1 000 Iriiri Poe coann pesna Comeletadt 221 wi Bloch Slat dac Eltoni Jnuedi Processing ICongiied 221 m Base Saracen 227 wi Fisctoraten Vetme 3 mi titer 01 nd Net tube pnag Prcessnct 223 mi Factoruten Volume 3 mi hber 01 int Nest sie Comet Z mi Lat Aide tiled Pase femund Process H ni Last ube fed Pana Compan 2 23 wt Erd Boch faved Pocenongt Cometeted 2 22 m loch
17. method run I gt Note An ended method cannot be continued permanently end the run click the End icon a i Note When ending a method run in advance it is possible to save the partial result 106 Getting Started with KTA avant 29 0351 83 AA 7 Runamethod 7 1 Before you start Warnings concerning use of hazardous substances WARNING Hazardous chemicals during run When using hazardous chemi cals 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 fol lowed by a neutral buffer and finally distilled water before service and maintenance Getting Started with AKTA avant 29 0351 83 AA 107 7 Runamethod 7 2 Applying the sample 7 2 Applying the sample Introduction 108 A number of different sample application techniques are available The sample can be applied either directly onto the column using the Sample pump or via a loop A loop can be filled either manually or by use of the Sample pump This section describes sample application using a syringe to manually fill a capillary loop The two stages of the sample application are described in the table below For detailed instructions and information regarding the different sample application
18. path before a run Illustration of the flow path The illustration below shows the flow scheme as illustrated in the System Control module The individual instrument modules are presented in the table below SyP to Col Syr to Loop to W Column 4 PreC 0 00 MPa DeltaC 0 00 MPa UV 1 0 27 MAU 0 00 mS cm pH 0 00 Sample Inlet Buffer 5 Inlet B B 1 Part Description 1 Sample inlet valve Getting Started with AKTA avant 29 0351 83 AA 73 6 Prepare the system for a run 6 2 Prepare the flow path Part Description 2 Sample pump 3 Inlet valve A 4 System pump A 5 Quaternary valve 6 Inlet valve B 7 System pump B 8 Injection valve 9 Capillary loop or Superloop 10 Column valve 11 Column 12 UV monitor 13 Conductivity monitor 14 pH valve with pH monitor and Flow restrictor 15 Outlet valve 16 Fraction collector Tubing and connectors The table below shows what types of tubing and connectors to use for different purposes AKTA avant 25 Scope ofuse Tubing Connector Photo Inlet tubing FEP o d 1 8 Tubing connector i d 1 6 mm 5 16 Ferrule yellow Tubing tocon PEEK o d Fingertight connector nect columns 1 16 i d 0 50 1 16 mm 74 Getting Started with AKTA avant 29 0351 83 AA 6 Prepare the system for a run 6 2 Prepare the flow path Scope ofuse Tubing Connector O
19. peak table Step Action 1 Open the Evaluation module and open a result Getting Started with AKTA avant 29 0351 83 AA 135 8 Evaluate and print the results 8 3 Print the results Step Action 2 Click the Print icon Result The Print Chromatograms dialog opens Print Chromatograms Printer Microsoft Office Document Image Writer aN Print Format Lo B U B E Chromatograms in each column Chromatograms in each row Use thick lines 3 In the Print Chromatograms dialog e Select Printer and Print Format e Click the Preview button Result A preview of the chromatogram displays in the Customize Report window 4 Are you satisfied with the layout e lf Yes continue to step 5 e If No select File Exit to return to the Print Chromatograms dialog and choose another print format 136 Getting Started with AKTA avant 29 0351 83 AA 8 Evaluate and print the results 8 3 Print the results Step Action 5 In the Customize Report window click the Print icon Result The Print dialog opens Printer Microsoft Office Document Image Writer Y Properties Print Range Copies all O Pages Enter page numbers and or page ranges seperated by commas For example 1 3 5 23 Number of copies 1 6 In the Print dialog select Printer from the list and click OK Result Your chromatogram is printed Print a report Follow the instruction below to print a standard f
20. position to use e which buffers and samples to prepare e which sample application technique to use e that the pH electrode is connected if applicable e which cassettes with corresponding deep well plates and or tubes to use in the Fraction collector if applicable e if itis a reversed phase chromatography RPC run WARNING Fraction collector Do not fractionate flammable liquids When running RPC methods or other procedures using organic sol vents collect fractions through the Outlet valve Getting Started with AKTA avant 29 0351 83 AA 71 6 Prepare the system for a run 6 1 Before you prepare the system WARNING RPC runs with 100 acetonitrile in AKTA avant 25 Always replace the green PEEK tubing between the used system pump and the pump pressure monitor with orange PEEK tubing i d 0 5 mm before running RPC with 100 acetonitrile Set the System pressure alarm to 10 MPa RPC runs with 100 acetonitrile in AKTA avant 150 Always replace the beige PEEK tubing between the used system pump and the pump pressure monitor with orange PEEK tubing i d 0 5 mm before running RPC with 100 acetonitrile 72 Getting Started with AKTA avant 29 0351 83 AA 6 Prepare the system for a run 6 2 Prepare the flow path 6 2 Prepare the flow path Introduction The flow path contains tubing valves pumps and monitors This section gives an overview of the flow path and describes how to prepare the flow
21. the Manual instructions dialog e Select Flow path Column position e Choose the Position used for connection of the column in this example Position 2 e Choose the Flow Direction Down flow Manual instructions System3 Instructions us a Parameters for Column position Pumps and pressures E Flow path Injection valve Position 2 sy Vale Flow Direction Inlet A Inlet B Down flow O Up flow pH valve Sample inlet Outlet valve Injection mark Monitors Fraction collection v Save result as 2 V Auto update parameter fields will be updated during method run click E Result The Column valve switches to position 2 7 In the Manual instructions dialog e Select Pumps and pressure System flow e Enter a low Flow rate e g 0 2 ml min Manual instructions System3 Instructions P a a T arameters for System flow Pumps and pressures A Flow rate 10 000 25 000 System flow ee 2 Sample flow 02 ml min Gradient Pump wash System wash Off Wash settings BufferPro pH Pressure control parameters How path Monitors Fraction collection v Pressure control Save result as 7 Auto update parameter fields will be updated during method run Click Ese Result A system flow of 0 2 ml min starts Getting Started with KTA avant 29 0351 83 AA 91 6 Prepare the system for a run 6 4 Connect a column Step Action
22. the purge con nector Open the purge valve by turning it counter clockwise about 3 quarters of a turn Draw a small volume of liquid slowly into the syringe with a rate of about 1 ml per second Getting Started with AKTA avant 29 0351 83 AA 87 6 Prepare the system for a run 6 3 Prime buffer inlets and purge system pumps Step Action 5 Close the purge valve by turning it clockwise Disconnect the syringe and discard its contents 6 Connect the syringe to the purge valve on the right pump head of System pump A and repeat steps 5 6 Keep the system flow running End the run Click the End icon in the System Control toolbar to end the run Lau 88 Getting Started with AKTA avant 29 0351 83 AA 6 Prepare the system for a run 6 4 Connect a column Connect a column Introduction This section describes how to connect a column to the instrument using a column holder and without introducing air into the flow path Several types of column holders are available for AKTA avant WARNING To avoid exposing the column to excessive pressure make sure that the pressure limit is set to the specified maximum pressure of the column Before connecting a column to the AKTA avant instru ment read the instructions for use of the column Methods automatically include a pressure alarm based on the specifications of the chosen column type However when running manual runs you have to set the pressure limits
23. total area Peaktable F C Percent of total peak area Peaktable G Height Peaktable H Width Peaktable4 Width at half height PeaktableJ Sigma Peaktable K Fraction tube id Peaktable L Faction ube N Peaktable M Type of peak limits Peaktable N Peak endpoint heights S Filter Peaks Fill settings Height less than 0 Fill peaks Width less than Pattern Area less than Width more than C Peak must be one of 20 largest C Do not show integration summary C Apply to all chromatograms Study peak characteristics The peaks in the integrated chromatogram are automatically labeled with their retention values The Peak Table is calculated using the same base unit ml CV or min as selected in the chromatogram Follow the instruction below to find the peak of interest Step Action 1 Find the retention value above a peak in the Chromatogram 2 Search the Peak Table for the same retention value 3 On the same row find the property of interest e g Area Peak data Integration summary Peak Retention Area of total of total Height name ml ml m amp U area peak area m4U 31 93 206 2683 26 47 47 39 164 965 3358 941728 1208 21 64 188 497 33 84 121 6052 15 60 2794 228 321 34 78 131725 1 6
24. valve 9 System pump B 19 Holder rails 10 Pressure monitor of System pumps Getting Started with AKTA avant 29 0351 83 AA 43 4 The instrument and software 4 2 UNICORN software overview 4 2 UNICORN software overview Introduction This section gives a brief overview of the UNICORN software a complete package for control supervision and evaluation of chromatography instruments and purification runs It also describes how to access the help utility that is included in UNICORN UNICORN modules overview UNICORN consists of four modules Administration Method Editor System Control and Evaluation The main functions of each module are described in the table below Module Main functions Administration Perform user and system setup system log and database administration Method Editor Create and edit methods System Control Start view and control runs Evaluation Open results evaluate runs and create reports Enter a UNICORN module To enter a module e click the Taskbar button of the module of interest r Administration EF Evaluation 53 System Control i Method Editor or e choose the module of interest in the Tools menu in any of the other software modules The illustration below shows the Tools menu of the Evaluation module i Column Handling Ctrl L Administration Method Editor System Control Options Change Passwords 4
25. 0351 83 AA 1 Introduction 1 1 Important user information e you understand the concepts of liquid chromatography e you have read and understood the Safety instructions chapter in the user documen tation e the instrument and software are installed configured and calibrated according to AKTA avant Installation Guide e a user account has been created according to UNICORN 6 Administration and Technical Manual Pushing the limits beyond Getting Started Users who are familiar with the instrument and software and want to learn more about the system s advanced features should refer to the list below To find out more please read about installation AKTA avant Installation Guide administration of UNICORN 6 Administration and Technical Manual databases instrument modules and KTA avant User Manual functions calibration AKTA avant User Manual predefined methods UNICORN 6 Method Manual column handling UNICORN 6 Method Manual manual method editing UNICORN 6 Method Manual BufferPro automatic UNICORN 6 Method Manual buffer preparation scouting UNICORN 6 Method Manual Design of Experiment UNICORN 6 Method Manual evaluation UNICORN 6 Evaluation Manual peak integration UNICORN 6 Evaluation Manual customized print format UNICORN 6 Evaluation Manual Getting Started with AKTA avant 29 0351 83 AA 7 1 Introduction 1 1 Important user information Intend
26. 0351 83 AA 117 7 Runa method 7 4 Monitor the run Tab Customization Curves Select curves to be displayed Curve style and color Customize the appearance of the displayed curves X axis Select base unit time or volume and set X axis scale Y axis Select which Y axes to display and set Y axis scale for the different curves e Click OK to close the dialog Note These operations determine which curves are displayed and their appearance in the Chromatogram pane and on printouts The original raw data curves can never be modified or removed from the result View details in Chromatogram Follow the instructions below to view selected parts of the chromatogram If you want to then identify a curve position the mouse pointer over the curve of interest Result The curve name is displayed select which curve the click the curve of interest Y axis scale refers to Result The Y axis scale changes to the unit applicable for the curve zoom in the curves select the area of interest by moving the pointer with the left mouse button held down Result The selected area is enlarged zoom out again click the right mouse button and select Reset Zoom Result The original area is restored in one step Customize Run Log The Run Log pane displays all registered actions during the run Scroll up to see the entire log To change what items to display in the pane 118 Get
27. 1 Prime the inlets fill the buffer inlets with liquid 2 Purge the System pumps remove air from the pump heads Overview The procedure consists of the following stages Prime all inlet tubing B to be used during the run Prime all inlet tubing A to be used during the run Prepare the system before purging the System pumps 1 2 3 4 Purge System pump B 5 Purge System pump A 6 End the run Prime inlet tubing B Follow the instructions below to fill all B inlet tubing to be used in the run with appropriate buffer solution Step Action 1 Make sure that all B inlet tubing that is to be used during the method run is immersed in the correct buffers Getting Started with AKTA avant 29 0351 83 AA 81 6 Prepare the system for a run 6 3 Prime buffer inlets and purge system pumps Step Action 2 In the Manual instructions dialog e Select Flow path Inlet B e Select the Position of the inlet to be filled from the drop down list Start at the inlet position with the highest number and end at the inlet position with the lowest number Manual instructions System3 Selected column type Select Instructions i Parameters for Inlet B 4 Pumps and pressures ad Position Flow path i Injection valve B1 s Column position Inlet pH valve Sample inlet Outlet valve Injection mark Monitors Fraction collection v Save result as Browse V Auto update of p
28. 2 Peak integration Illustration of Chromatogram and Peak Table The illustration below shows a Chromatogram pane including a Peak Table i Chrom 1 UV 1_280_Chrom 1 id101b 001 Cone B_Chrom 1 id101b 001 UV 1_280 01 BASEM Edited 1 _Chrom 1 Run 1 33 84 220 210 200 490 33 j 8 180 170 160 150 140 120 110 A UV 1_280 01 PEAK Edited Peak data Integration summary No Peak name Retention ml Area ml m4U Height m4U a 31 93 206 2683 164 965 2 33 58 94 1728 188 497 3 33 84 121 6052 225 321 4 34 78 13 1725 6 747 Customize Peak Table By default each peak in the Peak Table is displayed with its retention time area and height Follow the instruction below to display other peak data e Click the Customize icon to open the Customize dialog FE y 4 e Under the Peak Table tab Select which data to display in the peak tables from the Select peak table columns list 132 Getting Started with AKTA avant 29 0351 83 AA 8 Evaluate and print the results 8 2 Peak integration Click OK to display the settings and close the dialog Customize Chrom 1 Curve Style and Color Edit Texts Layout Library Header CurveNames Y Ass XAxis Cuve Peak Table Select table to display Select peak table columns a Peak name Peaktable A v Retention Peaktable B A Peaktable C hoe endpoint retentions A iea Peaktable D Peaktable E Percent of
29. 3 303 6747 Getting Started with AKTA avant 29 0351 83 AA 133 8 Evaluate and print the results 8 2 Peak integration Save results 134 Follow the instruction below to save your changes If you want to then save your changes to the original result click the Save icon in the toolbar g save the changed result as a new result e select File Save As to open the Save Result As dialog e Inthe Save Result As dialog select the location for the new result write a name for the new result click Save Getting Started with AKTA avant 29 0351 83 AA 8 Evaluate and print the results 8 3 Print the results 8 3 Print the results Introduction This section describes how to print a chromatogram and a standard format report UNICORN uses the printers and printer settings that are installed on your computer Customize the view Before printing make sure that the view is adjusted to show what is needed in the re port printout Customize the chromatogram and or peak table and view details in the chromatogram according to the instructions above Before printing check that e correct curves are displayed e correct data are displayed in the peak table e a suitable scaling of the axes has been selected e asuitable zoom factor has been selected Print a chromatogram and a peak table Follow the instruction below to print a chromatogram and if peak integration has been performed the related
30. 4 Getting Started with AKTA avant 29 0351 83 AA Access the help utility 4 The instrument and software 4 2 UNICORN software overview A comprehensive help utility is included in the UNICORN software The table below de scribes how to access the different parts of the help utility If you want to find information about a UNICORN module select Help Help for in the UNICORN module of interest Contextual Help Fi Instruction Set Golin GIF About UNICORN find information about e press the F1 key with the item of interest selected and the item currently select in focus ed and in focus e g a pane a dialog or a Note method phase To find information about a phase make sure that the Phase Properties tab is selected before pressing the F1 key or click the Help icon in the open dialog navigate the online help select Help Help for in any of the UNICORN modules see illustration above in the TOC Table of contents pane expand the headings of interest to navigate the content structure click the heading of interest to open a section search for a specific term in the online help select Help Help for in any of the UNICORN modules see illustration above in the Search pane enter the term of interest in the input field click the Search button Getting Started with AKTA avant 29 0351 83 AA 45 4 The instrument and software 4 2 UNICORN software overview
31. A avant 29 0351 83 AA 109 7 Runamethod 7 2 Applying the sample How to fill a capillary loop Follow the instruction below to fill the capillary loop with sample Step Action il 2 4 Connect a suitable capillary loop to Injection valve ports LoopF fill and LoopE empty Fill a syringe with sample Connect the syringe to the Injection valve port Syr Load sample into the capillary loop To avoid sample loss due to siphoning leave the syringe in the port until the sample has been injected onto the column during the run Tip It is recommended to overload the loop to make sure that the loop is com pletely filled Excess of sample will leave the valve through port W1 110 Getting Started with AKTA avant 29 0351 83 AA 7 Runa method 7 3 Start a method run 7 3 Start amethod run Introduction This section describes how to start a run using a previously created method If Column Logbook was enabled during installation of the software registration and selection of individual columns is possible at method start The Column Logbook function includes for example logging of column history For further information on column handling please refer to UNICORN 6 Method Manual Choose and start a method The instruction below describes how to open a method and start a run Step Action 1 Open the System Control module and click the icon Open Method Navigator ima Result The Method
32. AU 0 00 mS cm 0 00 120 Getting Started with AKTA avant 29 0351 83 AA 7 Runa method 7 5 After run procedures 7 5 After run procedures Introduction This section describes how to clean the instrument and columns after a chromatographic run and how to prepare the system for storage The instrument and the columns should be cleaned between the runs This will prevent for example sample contamination protein precipitation and column clogging If the instrument is not going to be used for a couple of days or longer the instrument columns and the pH flow cell should be filled with storage solution For further information about cleaning and maintenance procedures see the User Manual Tip To clean and fill the instrument and columns with storage solution use System CIP and Column CIP either as separate predefined methods or as phases in cluded in a chromatographic method 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 System cleaning After a method run is completed perform the following e Rinse the instrument with one or several cleaning solution s e g NaOH buffer solu tion or distilled water using System CIP e Empty the Fraction collector e Clean all spills on the instrument and on the bench using a moist tissue e Empty the waste vessel e
33. Check that the pH electrode is in appropriate buffer System storage If the instrument is not going to be used for a couple of days or longer also perform the following e Fill the system and inlets with storage solution e g 20 ethanol using System CIP Getting Started with AKTA avant 29 0351 83 AA 121 7 Runamethod 7 5 After run procedures Column cleaning After a method run is completed perform the following e Clean the column with one or several cleaning solution s using Column CIP Column storage If the column is not going to be used for a couple of days or longer also perform the following e Fill the column with storage solution e g 20 ethanol using Column CIP pH electrode storage Follow the instructions below to fill the pH flow cell with storage solution The calibration function is used to switch the position of the pH valve However no calibration is per 122 formed Step Action 1 Open the System Control module and select System Calibrate Result The Calibration dialog opens Calibration System3 Monitor to caibrate iii Curent value Calibration procedure Prepare for calibration Calibration for pH electrode buffer 1 Description pH for buffer 1 iaten te uing two pf eaibraton salons i can also be used to fill the pH cell with storage solution since the pH valve is in calibration position pH for buffer 2 For calibration preferably use pH standard buff
34. GE Healthcare Life Sciences Getting Started with AKTA avant Original instructions Table of Contents Table of Contents 1 Loc doc L116 ee Reed E etree Ure T E A denen E mre 5 1 1 Important user information 6 12 REQUICEORY TAPFORFCTION sssssssssssosssessesscorsecsasenossoseesscosoossocenssvsnsvoseossoseessecnsnrsbssbunscosebedenssevvssiestioreaseeny 10 Safety IMStrUCONS ceric cacet asst cco eee eatin esos ore cei 14 21 Safety DECC MS icsassscstcccsnsssessanasetbchavesdesvssises oosest eotscsustestisssssosaacceedatsaiiscletteadsnnannesdessstiatettocestatey 15 22 WCBS ienen naa A R E aN IR R 26 2 53 Emerg n y proc d reS nsiinansaniina a aR R RAE UR 29 24 Recycling informali Onasiesssanenman nana E RRN NAR 32 2 5 Declaration of Hazardous Substances DOHS csssecscsssssccsssssssssusssessessccssssssssusssessesecsssssenssees 33 SEGUE UD issis rsss rE E 36 The instrument and software e sessessssessssessesesessessssessesessesreesresessessesessssesessess 40 4 1 KTA avant instrument Overview eecssssssssssssssssssessssssesssssssssssssssssessssttersetterssstesrssttsrstttssettteedetteeee 41 4 2 UNICORN software overview 44 4 3 UNICORN software modules 47 43 1 Method Editor MOdule viccecceecccsssssssssessssssesssssssssssssssecssssssssssssssssssesssssuesssssusssssssssssssessssssuessssseeessane 48 4 3 2 System Control module 51 4 3 3 Evaluation module 53 Create Q Method eae een eee near
35. Navigator pane opens Method Navigator Open Methods Method Q E Folder name System A Eric 2 Methods Method1 System3 Method2 System3 Results Select the method to run and click the Run icon Ea pee Result The Start Protocol dialog opens Step through the displayed pages in the Start Protocol add requested input and make appropriate changes if necessary Click Next Getting Started with KTA avant 29 0351 83 AA 111 7 Runamethod 7 3 Start a method run Step Action 4 Click Finish on the last page of the Start Protocol Result e If column logging was chosen at installation of UNICORN and a column type was selected at method creation the Select Columns dialog opens Continue with steps 5 9 Select Columns System3 Enter or select a column individual Code Enter ID O Select ID 3288 1 Disable column logging for this run Apply to all methods with the same column type Methods Remark Column Barcode Column type Column Handling grun 1 0 Desalting Column Handling Scouting run 2 HiPrep 26 10 Desalting Column Handling Scouting run 3 HiPrep 26 10 Desalting e If column logging was not chosen at installation of UNICORN and or no column type was selected at method creation the run starts directly 5 Is the column to be used already registered e If No continue to step 6 e If Yes continue to step 9 6 In the Select Columns dialog click New Re
36. a 3 D a D fe 5 Getting Started with AKTA avant 29 0351 83 AA 101 6 Prepare the system for a run 6 7 Prepare for a run in a cold cabinet 6 7 Prepare for a run ina cold cabinet Introduction To fit the AKTA avant instrument in a cold cabinet the foldable door and pump cover can be removed For instructions see KTA avant User Manual When using the instrument in a cold room or cold cabinet make sure to follow the precautions listed below Precautions concerning runs in a cold cabinet NOTICE Avoid condensation If KTA avant is kept in a cold room cold cabinet or similar keep it switched on in order to avoid condensa tion NOTICE Avoid overheating If AKTA avant is kept in a cold cabinet and the cold cabinet is switched off make sure to switch off AKTA avant and keep the cold cabinet open to avoid overheating NOTICE Place the computer in room temperature If the KTA avant in strument is placed in a cold room use a cold room compatible computer or place the computer outside the cold room and use the Ethernet cable delivered with the instrument to connect to the computer Note When the instrument is kept in a cold room it is important to tighten all tubing connectors also the inlet manifold connectors Otherwise air might get into the flow path 102 Getting Started with KTA avant 29 0351 83 AA 6 Prepare the system for a run 6 7 Prepare for a run ina cold c
37. abinet Tip When runs are performed in a cold cabinet make sure to adjust the target temperature of the Fraction collector temperature control function The target temperature is 20 C by default Settings for the temperature control function can be edited in the System Settings dialog of System Control or in the Text Instructions pane in Method Editor Getting Started with AKTA avant 29 0351 83 AA 103 7 Runamethod 7 Runa method About this chapter This chapter describes how to start up and run a method and also how to handle the system after the run In this chapter This chapter contains the following sections Section See page 7 1 Before you start 105 7 2 Applying the sample 108 7 3 Start a method run 111 7 4 Monitor the run 116 7 5 After run procedures 121 104 Getting Started with AKTA avant 29 0351 83 AA 7 Runamethod 7 1 Before you start 7 1 Before you start Introduction Before starting a run it is necessary to read and understand the information in this section and to perform the checks listed below WARNING Always use appropriate personal protective equipment during op eration and maintenance of AKTA avant WARNING Hazardous substances When using hazardous chemical and bio logical 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 maint
38. adient used in the method is displayed in a Gradient illus tration 4 E 3 1 7 Oe oR vee maes h e eneen minenn v Coan fopa Camm vee oma a camma Semate Apgteaton Pesas ipee 050 MP FO2 200 Daliks m 2 Feast even 03 MPa 200 3 oa F Uwe fom memes Cohn Wash Fim Fate Une mn w Voden Cohn postion Asin 1 Moree oneg panpana Sna Wordlercihe 190 700 om Nom vane 1 000 sian L000 25000 wt 20 m mec k Cort te fom to pent ovea age Owe t Coiumn Wash ows yee ewer n X Use mauah pepaed tater oan wea a TEC E ratie pH montomng O Use asna tater apaan Butte Entin w soram siom heare Erea pam es ome a wae BEES prone an wired ty ee he ma cerporent he imn Logod m Method Editor toolbar icons The table below shows the Method Editor toolbar icons that are referred to in this Getting Started guide Function Function m New Method Opens the Open Method Navigator a New Method dialog where Opens the Method Naviga methods can be created tor where available meth ods are listed Getting Started with KTA avant 29 0351 83 AA 49 4 The instrument and software 4 3 UNICORN software modules 4 3 1 Method Editor module Icon id Function Save Saves the active method Icon Function Print Opens the Print dia log from where a method can be printed T Copy Copies the selected method or folder Paste Pastes a copied method or folder to a new
39. arameters during run Click L_Execute_ Result Inlet valve B switches to the selected port 3 Connect a 25 30 ml syringe to the purge valve of one of the pump heads of System pump B Make sure that the syringe fits tightly into the purge con nector 4 Open the purge valve by turning it counter clockwise about 3 quarters of a turn Draw liquid slowly into the syringe until the liquid reaches the pump 82 Getting Started with AKTA avant 29 0351 83 AA 6 Prepare the system for a run 6 3 Prime buffer inlets and purge system pumps Step Action 5 Close the purge valve by turning it clockwise Disconnect the syringe and discard its contents 6 Repeat steps 2 5 for each piece of inlet tubing B that is to be used during the run Prime inlet tubing A Follow the instruction below to fill all A inlet tubing to be used in the run with appropriate buffer solution Step Action 1 Make sure that all A inlet tubing that is to be used during the method run is immersed in the correct buffers 2 In the Manual instructions dialog e Select Flow path Inlet A e Select the Position of the inlet to be filled from the drop down list click Bate Result Inlet valve A switches to the selected port 3 Connect a 25 30 ml syringe to the purge valve of one of the pump heads of System pump A Make sure that the syringe fits tightly into the purge con nector 4 Open the purge valve by turning it counter clockwise about 3 qua
40. ariable list O Phase Method Setting 4 In the Print dialog e select Printer e click OK Result The method is printed Getting Started with AKTA avant 29 0351 83 AA 69 6 Prepare the system for a run 6 Prepare the system for a run About this chapter This chapter describes the preparations necessary to prepare the system before starting arun WARNING Always use appropriate personal protective equipment during op eration and maintenance of KTA avant system In this chapter This chapter contains the following sections Section See page 6 1 Before you prepare the system 71 6 2 Prepare the flow path 73 6 3 Prime buffer inlets and purge system pumps 81 6 4 Connect a column 89 6 5 Calibrate the pH monitor 94 6 6 Prepare the Fraction collector 96 6 7 Prepare for a run in a cold cabinet 102 70 Getting Started with KTA avant 29 0351 83 AA 6 Prepare the system for a run 6 1 Before you prepare the system 6 1 Before you prepare the system Introduction It is important to prepare the system in accordance with the settings in the method to be run Before preparing the system check the settings in the Method Editor and make sure that all accessories to be used are available Checklist Make sure to prepare the system in accordance with the settings in the method to be run Remember to check e which valve ports to use for inlets and outlets e which column type to use e which column
41. col Start Protocol Result The Start Protocol dialog opens Start Protocol Method Items to display at method start Fraction Collector Variable List Displays the fraction collector setup tube type Current setting item description and position Scouting Text Instructions Notes Gradient BufferPro Columns Evaluation Procedures Method Information System Settings Calibration Questions Changeable Batch ID V Result Name and Location Daremas In the Start Protocol dialog e Select items to display at method start Result Name and Location is selected by default e Click OK to confirm and close the dialog 6 Select the next phase in the Method Outline pane by clicking it and choose appropriate parameter values in the Phase Properties pane T Repeat step 6 until all phases are edited Tip If the instrument will not be used for a couple of days or longer add one or several System CIP phases to the end of the method as described below Getting Started with AKTA avant 29 0351 83 AA 61 5 Create a method 5 1 Guide to method creation 5 1 1 Create and edit methods Step Action 8 Click the Save the method icon a Result The Save As dialog opens al Methods Folder name System Last modified Createdby J UNICORN Database E Anita 01 14 2009 12 35 Defa
42. connecting the power cord Protective ground The AKTA avant instrument must always be connected to a grounded power outlet Power cord Only use grounded power cords delivered or ap proved by GE Healthcare 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 Installing the computer The computer should be installed and used according to the instructions provided by the manufactur er of the computer CAUTION Make sure that the waste vessels will hold all the produced volume of the run For AKTA avant 25 a suitable waste vessel should typi cally have a volume of 2 to 10 liters For AKTA avant 150 a waste vessel should have a volume of 40 liters Getting Started with AKTA avant 29 0351 83 AA 19 2 Safety instructions 2 1 Safety precautions NOTICE To avoid damage to KTA avant or other equipment when installing or moving the instrument follow the instructions below e The maximum level of the waste vessel for the waste tubing from the valves must be lower than 30 cm above the lab bench e The maximum level of the waste vessel for the waste tubing from the Fraction collector and the Buffer tray must be lower than the bench height e Vents on the KTA avant instrument To ensure adequate ventilation keep papers and other objects away from the vents of the
43. cumen tation Disconnect power Always switch off power to the instrument before replacing any component on the instrument unless stated otherwise in the user documentation Hazardous chemicals during maintenance When using haz ardous chemicals for system or column cleaning wash the system or columns with a neutral solution in the last phase or step NOTICE To avoid damage to AKTA avant or other equipment when perform ing maintenance on the AKTA avant instrument follow the instruc tions below Cleaning Keep the AKTA avant instrument dry and clean Wipe regularly with a soft damp tissue and if necessary a mild cleaning agent Let the AKTA avant instrument dry completely before use Advanced maintenance Read the instruction carefully before disassembly of the pump head Getting Started with AKTA avant 29 0351 83 AA 25 2 Safety instructions 2 2 Labels 2 2 Labels Introduction This section describes the safety labels and labels concerning hazardous substances that are attached to the AKTA avant instrument For information about marking of the computer equipment refer to the manufacturer s instructions Labels on the AKTA avant instrument The illustrations below show the labels that are attached to the AKTA avant instrument 26 Getting Started with AKTA avant 29 0351 83 AA Safety symbols 2 Safety instructions 2 2 Labels The following safety symbols are used in the label
44. e User Name field FI Log On UNICORN O Use Windows Authentication User Name Password Domain Access Group Eric Administrators v e click the Options button and select which UNICORN modules to start Start UNICORN Administration C System Control e click OK Met hod Editor O Evaluation Result The selected UNICORN modules open 7 Administration i gt System Control amp Method Editor s P User OA P i g lg UNICC J g Connect to system Follow the instructions below to connect the instrument to UNICORN 38 Getting Started with KTA avant 29 0351 83 AA 3 Start up Step Action al In the System Control module click the Connect to Systems icon Result The Connect to Systems dialog opens Connect to Systems Connected systems 1 selected max 3 System name Control View E System E System2 E System3 E Systems E B Systems E Systems E System 2 In the Connect to Systems dialog e Select a system e Select Control mode e Click OK Result The selected instrument can now be controlled by the software Getting Started with AKTA avant 29 0351 83 AA 39 4 The instrument and software 4 The instrument and software About this chapter This chapter gives an overview of AKTA avant instrument software and accessories Illustration of the system The illustrati
45. e both pump heads of System pump B Step Action 1 In the Manual instructions dialog e Select Pumps and pressures Gradient e Set Target to 100 B and Length to 0 min Manual instructions System3 Selected column type l _ Select Parameters for Gradient Pumps and pressures a Target 0 0 100 0 System f a ___ __ alice a 100 0 2 xB Sample flow Length 0 00 100000 0 Pump wash SS a System wash L 0 00 min Quaternary start concentrations Quatemary gradient BufferPro pH Column packing flow amp Flow path Monitors Instructions lt Browse Save resultas 2 V Auto update of parameters during run Click LExeeute_ Result Only System pump B is active 2 In the Manual instructions dialog e Select Flow path inlet B e Select the Position of one of the inlets to be used from the drop down list Manual instructions System3 Selected column type IL Select Parameters for Inlet B Pumps and pressures a Position Flow path Injection valve Bt Mi Column position Inlet A pH valve Sample inlet Outlet valve Injection mark Monitors Fraction collection Instructions lt __ Browse Save result as 2 V Auto update of parameters during run Click LEwecute J Result Inlet valve B switches to the selected port Getting Started with AKTA avant 29 0351 83 AA 85 6 Prepa
46. e tabs described in the table below Getting Started with KTA avant 29 0351 83 AA 127 8 Evaluate and print the results 8 1 View the results Tab Customization Curves Select curves to be displayed Curve style and color Customize the appearance of the displayed curves X axis Select base unit time CV or volume and set X axis scale Y axis Select which y axes to display and set Y axis scale for the different curves Header Select which parameters variables questions and or notes to display in the header information at top of the chromatogram e Click OK to save the changes and close the dialog Note These operations determine which curves are displayed and their appearance in the Chromatogram pane and on printouts The original raw data curves can never be modified or removed from the result View details in Chromatogram Follow the instructions below to view selected parts of the chromatogram If you want to then identify a curve position the mouse pointer over the curve of interest Result The curve name is displayed select which curve the Y axis scale refers to click the curve of interest Result The Y axis scale changes to the unit applicable for the curve zoom in on the curves select the area of interest by moving the pointer with the left mouse button held down Result The selected area is enlarged zoom out again click the right mouse
47. ectrode Maintenance when required The following maintenance should be performed by the user of KTA avant when required Maintenance action Clean the instrument externally Perform System CIP Perform Column CIP Clean the Fraction collector Replace tubing and connectors Storage of pH electrode Clean the pH electrode Clean the Conductivity flow cell Calibrate the Conductivity monitor Calibrate the UV monitor Replace Mixer Getting Started with KTA avant 29 0351 83 AA 141 9 Maintenance Maintenance action Replace o ring in Mixer Replace the UV flow cell Replace the Flow restrictor Replace inlet filters Clean the check valves Replace check valves Replace pump piston seals Replace pump pistons Replace pump rinsing system tubing Replace valve modules 142 Getting Started with KTA avant 29 0351 83 AA 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 imagination at work GE imagination at work and GE monogram are trademarks of General Electric Company AKTA AxiChrom and UNICORN are trademarks of GE Healthcare companies Windows is a trademark of icrosoft Corporation 2009 2012 General Electric Company All rights reserved First published Dec 2012 All
48. ed by recipe bor mubicorpormeet tasters Cohen Logos Enatie gong of O Cotsen Patoma Tout oor Duaston k Varabies Getting Started with AKTA avant 29 0351 83 AA 59 5 Create a method 5 1 Guide to method creation 5 1 1 Create and edit methods 60 Step Action 4 In the Phase Properties pane of the Method Settings phase click the Result Name amp Location button to specify the name and location of the results from the method runs Result Name amp Location Result The Result Name amp Location dialog opens Result Name amp Location No Result Result location l DefaultHome Folder name for Design of Experiments or Scouting Result name O Name O Variable Method name O Date Add unique identifier In the Result Name amp Location dialog e Set Result location by clicking the Browse button and select a folder in which to save the results By default the results will be saved in your home folder e Select Result name With the default selection Method name the results will be named with the method name together with a serial number e Click OK to confirm and close the dialog Getting Started with AKTA avant 29 0351 83 AA 5 Create a method 5 1 Guide to method creation 5 1 1 Create and edit methods Step Action 5 In the Phase Properties pane of the Method Settings phase click the Start Protocol button if you wish to include a start proto
49. ed use AKTA avant is a liquid chromatography system intended for process development The system can be used to screen for optimal choice of columns media and running param eters to purify selected proteins The AKTA avant system is 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 Cautions CAUTION CAUTION indicates a hazardous situation which if not avoided could result in minor or moderate injury It is important not to pro ceed until all stated conditions are met and clearly understood Notices NOTICE 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 8 Getting Started with KTA avant 29 0351 83 AA 1 Introduction 1 1 Important user information Tip A tip contains useful information that can improve or optimize your procedures Typographical conventions Software items are identified
50. ed with AKTA avant 29 0351 83 AA 2 Safety instructions 2 1 Safety precautions CAUTION To avoid hazardous situations when operating the AKTA avant system follow the instructions below e Risk of explosion Do not use Mixer chamber 15 ml in KTA avant 25 The maximum pressure for Mixer chamber 15 ml is 5 MPa e Risk of breaking test vials Do not use excessive force to press vials with incorrect dimensions into the Fraction collector cas settes Glass vials may break and cause injuries e Hazardous chemicals or biological agents in UV flow cell Make sure that the entire flow cell has been flushed thoroughly with bacteriostatic solution e g NaOH and distilled water before service and maintenance e pH electrode Handle the pH electrode with care The glass tip may break and cause injury e Max weight on Buffer tray Do not place containers with a volume of more than 10 liters each on the Buffer tray The total allowed weight on the Buffer tray is 40 kg e Max size of bottles on front panel Do not fasten bottles with a volume of more than 1 liter in the front panel rails e Risk of explosion Do not use Mixer chamber 15 ml in KTA avant 25 The maximum pressure for Mixer chamber 15 ml is 5 MPa Getting Started with AKTA avant 29 0351 83 AA 23 2 Safety instructions 2 1 Safety precautions NOTICE To avoid damage to KTA avant or other equipment when operating the instrument follow the inst
51. elect File Exit UNICORN Log off Eric Exit UNICORN Result All open UNICORN modules close Note If an edited method or result is open and not saved when you try to exit or log off UNICORN you will see a warning Click Yes to save No to exit without saving or Cancel to stay logged on Method Editor A Do you want to save the changes made to Method 1 Shut down the instrument Switch off the instrument by pressing the Power switch to the O position 124 Getting Started with KTA avant 29 0351 83 AA 8 Evaluate and print the results 8 Evaluate and print the results About this chapter This chapter describes how to use the Evaluation module to evaluate and print the results of a run For further information please refer to UNICORN 6 Evaluation Manual In this chapter This chapter contains the following sections Section See page 8 1 View the results 126 8 2 Peak integration 130 8 3 Print the results 135 Getting Started with AKTA avant 29 0351 83 AA 125 8 Evaluate and print the results 8 1 View the results 8 1 Introduction A result holds a complete record of the run including method system settings chro View the results matogram and run log This section describes how to view a result and how to change the view settings in the Evaluation module Open a result Follow the instruction below to open a result 126 Step Action 1 Open the E
52. enance and decommissioning of the equipment WARNING High pressure AKTA avant operates under high pressure Wear protective glasses at all times Checklist Make sure that the system is correctly prepared Check that e The system is prepared according to the settings in the method to be run e Asuitable column has been selected for the application consider target protein and pressure range e The buffer inlet tubing is immersed in correct buffer vessels consider solution iden tity and volume e All waste tubing is immersed in appropriate waste vessels consider vessel size placement and material e No tubing is twisted and the flow path is free from leakage Getting Started with AKTA avant 29 0351 83 AA 105 7 Run a method 7 1 Before you start Hold pause or stop the run At the end of a method the run stops automatically All pumps stop and an acoustic end signal sounds and End is displayed in the Run Log To interrupt a method during a run you may use the Hold Pause or End icons in System Control A held or paused method run can be resumed by using the Continue icon See the instructions in the table below If you want to then temporarily hold the method with current click the Hold icon flow rate and valve positions sustained temporarily pause the method and stop click the Pause icon all pumps il resume for example a held or paused click the Continue icon
53. er as separate predefined methods or as phases included in chromato graphic methods Tip The System CIP and System Preparation phases are designed to use one cleaning solution each The System Preparation method includes two System Preparation phases and the System CIP method includes three System CIP phases To use additional solutions sequentially add phases to the method Note Usually 0 5 1 M NaOH is used for cleaning of the system and column However before choosing cleaning agent always consider the media and buffers used and also the chemical resistance of the column Getting Started with KTA avant 29 0351 83 AA 5 Create a method 5 1 Guide to method creation 5 1 2 Predefined methods 5 1 2 Predefined methods The predefined methods that are available are described in the table below Method Description Affinity Chro matography AC After equilibration and sample application the protein of interest is adsorbed to the column ligand After a wash to remove un bound sample elution is performed either by using a buffer containing a competitor to displace the protein of interest or by changing the pH or ionic strength Finally the column is re equilibrated with start buffer Anion Exchange After equilibration and sample application negatively charged Chromatography proteins are adsorbed to the column ligand After a wash to re AIEX move unbound sample elution is performed using a gradient of
54. ers PH 4 or 7 as the first calibration point and a pH standard buffer close to the lowest or highest pH Last calibrated on 2009 01 13 10 52 07 01 00 you need to measure as your second point Calibrated electrode slope Parameters Assymetry potential at pH 7 mV Ha hel ee ul 2 In the Calibration dialog select pH from the Monitor to calibrate drop down list 5 i Prepare for calibration Press Result The pH valve switches to the calibration position 4 Prepare at least 10 ml storage solution by mixing equal volumes of a stan dard buffer pH 4 and a 1 M Potassium Nitrate KNO3 solution Getting Started with AKTA avant 29 0351 83 AA 7 Runa method 7 5 After run procedures Step Action 5 Filla syringe with approximately 10 ml of the storage solution Connect the syringe to the pH valve port Cal and inject the storage solution Press cose Result The pH valve switches back to the default position and the Calibration dialog closes No calibration is performed Log off or exit UNICORN Follow the instruction below to log off or exit UNICORN This can be performed from any of the UNICORN modules If you want to then log off UNICORN select File Log off 3 Log off Eric Exit UNICORN Result All open UNICORN modules close and the Log On dialog opens Getting Started with AKTA avant 29 0351 83 AA 123 7 Runamethod 7 5 After run procedures If you want to then exit UNICORN s
55. es Shattered glass from falling bottles may cause injury Spilled liquid may cause fire hazard and personal injury _ e Electrical shock hazard after spillage If there is a risk that large volumes of spilled liquid may penetrate the casing of the AKTA avant instrument immediately switch off the instrument disconnect the power cord and contact an authorized service engineer Getting Started with AKTA avant 29 0351 83 AA 21 2 Safety instructions 2 1 Safety precautions WARNING e Moving parts in Fraction collector Do not open the Fraction collector drawer when the Fraction collector is active If you need to access the Fraction collector press Pause and make sure that the movement has stopped before opening the drawer e Using a Superloop After loading a Superloop always plug the Syr port on the Injection valve with a Stop plug With a Super loop connected to the valve an over pressure may be created during injection e 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 mainte nance e Hazardous biological agents during run When using haz ardous 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 22 Getting Start
56. es a copied re sult or folder to the selected folder B E Undo Restores the result to the state it was in before the last change Report Opens the Create report dialog where a re port of the result can be created im View Documentation Opens the Documentation dialog that contains the complete documentation for a method run Customize Opens the Cus tomize dialog where curve settings peak table content and header content can be set Peak Integrate Opens the Peak Integrate dialog from where curves can be inte grated 54 Getting Started with KTA avant 29 0351 83 AA 5 Create a method 5 Create a method About this chapter A method is created in the Method Editor module This chapter describes method creation based on predefined methods and also how to print a method Please refer to UNICORN 6 Method Manual for more detail about method creation In this chapter This chapter contains the following sections Section See page 5 1 Guide to method creation 56 5 2 Print a method 68 Getting Started with AKTA avant 29 0351 83 AA 55 5 Create a method 5 1 Guide to method creation eral Guide to method creation About this section This section describes how to create a new method based on a predefined method and how to edit a method The section also provides an overview of the predefined methods and phases that are available For further information about
57. f equipment N hazardous materials in excess of the limits established by 207 the Chinese standard SJ T11363 2006 Requirements for This symbol indicates that the product might contain waste and must be collected separately Please contact an authorized representative of the manufacturer for in Concentration Limits for Certain Hazardous Substances in Electronic Information Product 28 Getting Started with AKTA avant 29 0351 83 AA 2 Safety instructions 2 3 Emergency procedures 2 3 Emergency procedures Introduction This section describes how to perform an emergency shutdown of the AKTA avant system This section also describes the results in the event of power failure or network interruption Emergency shutdown In an emergency situation follow the steps below to stop the run Step Action 1 Press the Pause button on the instrument display This will stop all pumps in the instrument To pause the run from UNICORN click the Pause icon Getting Started with AKTA avant 29 0351 83 AA 29 2 Safety instructions 2 3 Emergency procedures Step Action 2 If required switch off power to the instrument by pressing the Power switch to the O position The run is interrupted immediately Power failure The result of a power failure depends on which unit is affected Power failure to will result in AKTA avant instrument e The run is interrupted immediately e The data collected
58. goods and services are so d 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 repre sentative for the most current information UNICORN Any use of this software is subject to GE Healthcare Standard Software End User License Agreement Standard Software End User GE Healthcare Europe GmbH or Life Sciences Software Products A copy of this License Agreement is available on request Munzinger Strasse 5 D 791 GE Healthcare UK Limited Freiburg Germany Amersham Place Little Chalfont Buckinghamshire HP7 9NA UK GE Healthcare Bio Sciences Corp 800 Centennial Avenue P O Box 1327 Piscataway NJ 08855 1327 USA GE Healthcare Japan Corporation Sanken Bldg 3 25 1 Hyakuni ncho Shinjuku ku Tokyo 169 0073 Japan 29 0351 83 AA 03 2013
59. graphy runs Evaluation panes As illustrated below the Evaluation module contains three panes When a result is opened from the Result Navigator 1 the Chromatogram pane 2 is displayed After a peak integration has been performed details about the peaks are shown in the lower right hand pane 3 in the tabs Peak data and Integration summary In the Evaluation module it is also possible to view the complete documentation about the results and to generate reports feos Recers urn Fred Rents on Fam FY nents tents iil UA 700 Cheon taetae oat raD Creom a0 oe Fenton Orn t naa G vi U8 POOO BASEN Rates 1 Chron 1 Pn 1 Feide nave BJ UNCOAN Databare Buea Direi Pa ceSeusesssshtseees 88 38 6B Peak name Rewal Aespa Wega belly 1 ons wan mss 2 ne sams sar 3 nu me z5x 4 PEJ 1178 en Getting Started with AKTA avant 29 0351 83 AA 53 4 The instrument and software 4 3 UNICORN software modules 4 3 3 Evaluation module Evaluation toolbar icons The table below shows the Evaluation toolbar icons that are referred to in this Getting Started Icon Function Open Result Navigator Opens the Result Navigator where available results are listed Icon E Function Save Saves the changes made to the current result Print Opens the Print chromatograms dialog from where a chro matogram can be printed Copy Copies the selected result or folder Paste Past
60. h RLS List of hazardous substances and their concentrations PF mhasa SWaARmrRAnNamRree Indication for each major part if substance exceeds limit Value Meaning O Indicates that this toxic or hazardous substance contained in all of the homogeneous materials for this part is below the limit requirement in SJ T11363 2006 RRBAS A SMREBZAEM ASRS NSB ES T11363 2006 tr EAL TEN PRS RAT Indicates that this toxic or hazardous substance contained in at least one of the homogeneous materials used for this part is above the limit requirement in SJ T11363 2006 e Data listed in the table represents best information available at the time of publication RRBZASASMRSLSEAREHR HRPA THe RBH J T11363 2006 RHEL BY Re 5k UE RRP THEA Ac 7h AY PT BERR SAY Be fe fs 34 Getting Started with AKTA avant 29 0351 83 AA 2 Safety instructions 2 5 Declaration of Hazardous Substances DoHS Component Hazardous substance 0 x name BSAS MAMI 0 X EA E Pb Hg Cd Cr6 PBB PBDE Ho Ro m ZN OMB 2 RRA ZR AR AKTA avant 25 0 0 0 0 0 0 28 9308 42 AKTA avant 150 0 0 0 0 0 0 28 9763 37 1 The product has not been tested as per the Chinese standard SJ T11363 2006 Requirements for Concentration Limits for Certain Hazardous Substances in Electronic Information Product Getting Started with AKTA avant 29 0351 83 AA 35 3 Start up 5 Start up Introduction This chapter describe
61. 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 controls indicators and connections are identified in the text by bold text e g Power switch Text entries that UNICORN generates or that the user must type are represented by a monotype typefacele g Program Files GE Healthcare UNICORN bin UNI CORN Instrument Server exe config Getting Started with AKTA avant 29 0351 83 AA 1 Introduction 1 2 Regulatory information 1 2 Regulatory information Introduction This section describes the directives and standards that are fulfilled by AKTA avant Manufacturing information The table below summarizes the required manufacturing information For further infor mation see the EC Declaration of Conformity document Requirement Content Name and address of GE Healthcare Bio Sciences AB Bj rkgatan 30 SE 751 manufacturer 84 Uppsala Sweden CE conformity This product complies with the European directives listed in the table below by fulfilling the corresponding harmonized standards For further information see the EC Declaration of Conformity document Directive Title 2006 42 EC Machinery Directive MD 2006 95 EC Low Voltage Directive LVD 2004 108 EC ElectroMagnetic Compatibility EMC Directive 1999 5 EC Radio Equipment and Telecommunications Terminal Equipment R amp TTE Directive CE marking ce
62. instrument e Disconnect power To prevent equipment damage always switch off power to the AKTA avant instrument before an instru ment module is removed or installed or a cable is connected or disconnected e Misuse of UniNet 9 connectors The UniNet 9 connectors at the rear panel should not be mistaken for Firewire connectors Do not connect any external equipment to the UniNet 9 con nectors Do not disconnect or move the UniNet 9 bus cable 20 Getting Started with AKTA avant 29 0351 83 AA 2 Safety instructions 2 1 Safety precautions System operation WARNING To avoid personal injury when operating the AKTA avant system follow the instructions below e Rotating the instrument Make sure that there is always at least 20 cm of free space around the AKTA avant instrument to allow for sufficient ventilation and rotation on the swivel foot When rotating the instrument take care not to stretch or squeeze tubing or cables A disconnected cable may cause power interruption or network interruption Stretched tubing may cause bottles to fall resulting in liquid spillage and shat tered glass Squeezed tubing may cause increase in pressure or block liquid flow To avoid the risk of knocking over bottles always place bottles on the buffer tray and close the doors before rotating the instrument e Fasten sample bottles Always fasten bottles and cassettes to the rails at the sample tray Use appropriate holders for bottl
63. l not leak or mutate under normal operating conditions so that the use of such electronic information products will not result in any severe environmental pollution any bodily injury or damage to any assets The unit of the period is Year In order to maintain the declared EFUP the product shall be operated normally according to the instructions and environmental conditions as defined in the product manual and periodic maintenance schedules specified in Product Maintenance Procedures shall be followed strictly Consumables or certain parts may have their own label with an EFUP value less than the product Periodic replacement of those consumables or parts to maintain the declared EFUP shall be done in accordance with the Product Maintenance Procedures This product must not be disposed of as unsorted municipal waste and must be collected separately and handled properly after decom missioning Getting Started with AKTA avant 29 0351 83 AA 33 2 Safety instructions 2 5 Declaration of Hazardous Substances DoHS Label Meaning O BRERA AS MS ABWSI 11363 2006 HFRS RHAS ASYVRMORBER PREDARE EYA MEPHRMEAA FROPREAM RAR REL EAHA BBS RASREXSRRRE APERAS RANI BK RUiRAS OFiak SRS MR XAF ARIE PAW REAR EE PEW RAE MACHETE SER HP METS mE FEA PAE k R EA RIFER FMP HEFNER S BE ARS AMR ARS FARMS ARR ELEAF RASH REAR BRAS mn SE PRA LAS CREAT AA ANT Gn AY ER OR FA ABR ANF 6 EEA a RAN A AY ED SB EAS N
64. larm delta column pressure Enabled 3 50 MPa 0 00 MPa Completed 0 01 min Wavelength 280 rm Off rm Off nm Issued Processing Find Included entry types Included feedback types Set Mark Issued Block Start Processing Alarms Wamings and Enors Completed Method Failed Manual Ignored System Cancelled v v v v v v Adjust retention zero to injection number Base Time O Volume Getting Started with KTA avant 29 0351 83 AA 129 8 Evaluate and print the results 8 2 Peak integration 8 2 Peak integration Introduction Peak integration is used to identify and measure curve characteristics including peak areas retention times and peak widths This section describes how to perform peak in tegration using the UNICORN software For further information on peak integration please refer to UNICORN 6 Evaluation Manual Perform peak integration Follow the instruction below to integrate the curves in a result Step Action 1 Open the Evaluation module and open a result from the Result Navigator 2 Click the Peak Integrate icon Result The Peak Integrate dialog opens 130 Getting Started with AKTA avant 29 0351 83 AA 8 Evaluate and print the results 8 2 Peak integration Step Action 3 In the Peak Integrate dialog e Select which curve to integrate
65. llector is to be used make sure that tubing is connected between the Outlet valve Frac port and the Fraction collector and prepare the fraction collector Otherwise immerse the outlet tubing in suitable tubes or flasks Waste ports The table below shows the waste ports of Injection valve pH valve and Outlet valve Valves and ports Illustrations Injection valve Waste ports e W1 W2 pH valve Waste port e W3 Getting Started with AKTA avant 29 0351 83 AA 77 6 Prepare the system for a run 6 2 Prepare the flow path Valves and ports Illustrations Outlet valve o N Waste port e W Out9 Out6 Location of waste tubing All waste tubing is found on the rear of the instrument see the illustration below Part Description 1 Waste tubing from the Injection valve the pH valve and the Outlet valve W W1 W2 and W3 78 Getting Started with KTA avant 29 0351 83 AA 6 Prepare the system for a run 6 2 Prepare the flow path Part Description 2 Waste tubing from the Fraction collector and the Buffer tray Prepare the waste tubing Follow the instruction below to prepare the waste tubing Step Action 1 Make sure that the four pieces of waste tubing from the Injection valve the pH valve and the Outlet valve W W1 W2 and W3 are placed in a vessel placed below the bench NOTICE The maximum level
66. method creation see UNICORN 6 Method Manual In this section 56 This section contains the following subsections Section See page 5 1 1 Create and edit methods 57 5 1 2 Predefined methods 65 5 1 3 Predefined phases 67 Getting Started with AKTA avant 29 0351 83 AA 5 Create a method 5 1 Guide to method creation 5 1 1 Create and edit methods 5 1 1 Create and edit methods Introduction Method creation in UNICORN 6 is performed in the Method Editor module Predefined methods are available for a number of different chromatography techniques for example affinity chromatography and gel filtration There are also predefined methods for column preparation and system cleaning A method is comprised of a number of phases Each phase represents a major process step in the method for example equilibration or elution If desired a predefined method can be edited by adding deleting and or rearranging phases and also by editing the settings of each phase included in the method Illustration of the Method Editor user interface The illustration and table below show the Method Editor module and its panes Cohen oe Hho Cato ateme 1 et Meros Nore v Tien Popat Cot hae Os Conn Wee Samie Aoi aron Proinae int peccnn 0 PRR Da v D Poeman int datocen 030 Pai 00 3 Bata icv EE Use Mom pectic Cohn Wash Flom Rate Unt mn w bodine Cohen postion Potent Mren ar tenes seeen Ss Wavelength
67. n collector Follow the instructions below to prepare the Fraction collector before a run Cassettes and Cassette tray Step Action 1 If you are to use Cassettes with the QuickRelease function open the Cas settes 96 Getting Started with AKTA avant 29 0351 83 AA 6 Prepare the system for a run 6 6 Prepare the Fraction collector Step Action 2 Place the tubes and deep well plates in the Cassettes Make sure that the deep well plates are rotated so that the well marked A1 is positioned above the A1 marking on the Cassette 3 Getting Started with AKTA avant 29 0351 83 AA 97 6 Prepare the system for a run 6 6 Prepare the Fraction collector Step Action 4 6 98 Place the Cassettes on the Cassette tray Make sure that the Cassette type code see illustration below faces the front of the tray marked with the GE logo Open the Frac drawer by pressing the handle upwards and pulling out the drawer Place the Cassette tray on the Tray support of the Frac drawer Make sure that the front of the tray marked with the GE logo faces the front of the drawer and is hooked onto the two pins Getting Started with AKTA avant 29 0351 83 AA 6 Prepare the system for a run 6 6 Prepare the Fraction collector Step Action 7 Close the Frac drawer Make sure that it snaps into closed position 50 ml tube tray Step Action 1 Place 50 ml tubes in the 50 ml tube tray 2 Open the Frac dra
68. nd re equilibrated with start buffer Result The phases included in the chosen method is shown in the Method Outline pane and the default settings for each of the phases is shown in the Phase Properties pane 58 Getting Started with AKTA avant 29 0351 83 AA 5 Create a method 5 1 Guide to method creation 5 1 1 Create and edit methods Step Action 3 In the Phase Properties pane of the Method Settings phase edit general settings like Column type and Method Base Unit UNICORN automatically calculates correct settings for volume flow rate and pressure limits based on the selected column type The illustration below shows the Method Outline pane and the Phase Properties pane of the Method Settings phase hue ropert Test itna iT Mathod Senin Cohan vefechons Ment Nane t Locaton Shamte Clonee Coena Squtmbretion Stat Poca mh Method Notes z Coin wiae 0100 si Sample Application Precuse fet pechen 3000 MPa p02 2000 Aal ialis 3 Presnse int detacotmn 2000 MPa 002 20 0 SS _ EE Vere tow ostren Cohan Wash Fiw Rate Ut atin e Changeon Broar saa ines Wavelerephe 190 700 rm Pam isie 1 000 n n 49000 25 0004 wi 200 m v FF Conio he tow to avoid overprersure Owe Cohen Wash Ow im X Uso maruy ponpe badiasi wia at F wane er 3 E brate gh mentoring Equiibration O Ue Dulteb ro merate batter pancaran Erata as vores diam Hecoe Sirta Eiran D Samoe riet m 20 70 boconmendeti Coc Owin
69. nd recycled according to national and local environmental regulations CAUTION Always use appropriate personal protective equipment when decommissioning the equipment Disposal of electrical components Waste comprising electrical and electronic equipment must not be disposed of as un sorted municipal waste and must be collected separately Please contact an authorized representative of the manufacturer for information concerning the decommissioning of equipment 32 Getting Started with AKTA avant 29 0351 83 AA 2 Safety instructions 2 5 Declaration of Hazardous Substances DoHS 2 5 Declaration of Hazardous Substances DoHS Introduction The following product pollution control information is provided according to J T11364 2006 Marking for Control of Pollution caused by Electronic Information Products RHESI T11364 2006 BFS 5 Ri ITER BRAM PARR Bal 7m aS Symbols used in pollution control label tis a eS RHE HER Ze HL H Label Meaning D This symbol indicates the product contains hazardous materials in ex cess of the limits established by the Chinese standard SJ T11363 2006 Requirements for Concentration Limits for Certain Hazardous Sub stances in Electronic Information Products The number in the symbol is the Environment friendly Use Period EFUP which indicates the period during which the toxic or hazardous substances or elements contained in electronic information products wil
70. of the waste vessel for the waste tubing from the valves must be lower than 30 cm above the lab bench 2 Make sure that the three pieces of waste tubing from the Fraction collector and the Buffer tray are placed in a waste vessel placed below the bench NOTICE The maximum level of the waste vessel for the waste tubing from the Fraction collector and the Buffer tray must be lower than the bench height Getting Started with KTA avant 29 0351 83 AA 79 6 Prepare the system for a run 6 2 Prepare the flow path Step Action 3 Make sure that the waste tubing from the Fraction collector and the Buffer tray is cut to appropriate length It is important that the tubing is not bent and will not be submerged in liquid during the run Note Ifthe tubing is too short replace it with new tubing Do not lengthen the tubing as this might cause obstruction of the tubing and flooding in the Frac chamber CAUTION Make sure that the waste vessels will hold all the produced volume of the run For KTA avant 25 a suitable waste vessel should typi cally have a volume of 2 to 10 liters For KTA avant 150 a waste vessel should have a volume of 40 liters 80 Getting Started with KTA avant 29 0351 83 AA 6 Prepare the system for a run 6 3 Prime buffer inlets and purge system pumps 6 3 Prime buffer inlets and purge system pumps Introduction Before usage of the System pumps it is important to
71. oing method run in the System Control module The current system status is shown in the System state panel in the Run Data pane For example it may state Run Wash or Hold The same information is also shown on the Instrument display This section describes the data shown in System Control during a run the layout of the module and the procedure to customize the view of the different panes Illustration of the System Control user interface 116 In the System Control module four panes show by default see illustration and table below as fe O a OS e ease 5 DIT A cSuesessesase o 02 Pun Log Fater en 220 mi Sytem Mom 1 000 bined Pre coke permen rued Procera 22A wi Sysas Now 1 000 Linin Poe coann presne Koneletedt 22 mi Bloch Slat Aac Eloni Deiued Processrgl Comte 221 mi Bme SametuMan 2 27 mi Fisctonaten Volume 3 nt Abes 01 inf Net tube fiad Pocersng os 223 ni Fiaetoraten Volume 3 mi kbar 1 lid Nest Abe Completed 2 20 mi Last Abe tiled Pane faund Processing 2 20 ni Lact hbe fded Pama Completed End Boch pued Procersng JComcitec 222 m ioch Lren gadert Passed Pocesing Completed 222 mi Dave Semen 2 22 mi Grader 100 00 RP 10 00 ICV Proua Processing 2 22 mi Graders 100 00 0 10 00 CV Compete AZ mi Lat be filed The ayatam pall pane alter the delay woken buns been collected fiystead Meted murs c Lnear adere tao watch Connection Connected n contrat
72. on below shows the AKTA avant instrument with UNICORN software installed on a computer In this chapter This chapter contains the following sections Section See page 4 1 AKTA avant instrument overview 41 4 2 UNICORN software overview 44 4 3 UNICORN software modules 47 40 Getting Started with AKTA avant 29 0351 83 AA 4 The instrument and software 4 1 AKTA avant instrument overview 4 1 AKTA avant instrument overview Introduction This section shows an overview of the AKTA avant instrument Technical details about the instrument and the individual modules are found in the AKTA avant User Manual Exterior design AKTA avant has a modular design with all the liquid handling modules placed on the exterior of the instrument Buffer vessels are placed on the Buffer tray on top of the in strument An Instrument display is placed on the front From this side the built in Fraction collector is handled as well as the sample The rest of the modules are placed on the right hand side of the instrument This side can be covered by a foldable door and a pump cover By rotating the instrument using the swivel foot any side is easily accessed Operating ranges The table below shows some of the operational limits of the instruments AKTA avant 25 Parameter Limits Flow rate Up to 25 ml min Note When running the Column packing flow instruction the maximum flow rate for AKTA avant 25 is 50 ml min Ma
73. on the Control of Pollution Caused by Electronic Information Products China Restriction of Hazardous Substances RoHS 13 2 Safety instructions 2 Safety instructions About this chapter This chapter contains instructions of how to handle the KTA avant instrument in a safe way In this chapter This chapter contains the following sections Section See page 2 1 Safety precautions 15 2 2 Labels 26 2 3 Emergency procedures 29 2 4 Recycling information 32 2 5 Declaration of Hazardous Substances DoHS 33 14 Getting Started with AKTA avant 29 0351 83 AA 2 Safety instructions 2 1 Safety precautions 2 1 Safety precautions Introduction The AKTA avant instrument is 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 injuries or damage to the equipment The safety precautions in this section are grouped into the following categories e General precautions e Flammable liquids e Personal protection e Installing and moving the instrument e System operation e Maintenance Getting Started with AKTA avant 29 0351 83 AA 15 2 Safety instructions 2 1 Safety precautions General precautions WARNING Always follow these General precautions to avoid injury when using the AKTA avant instrument e Do not opera
74. ormot report Step Action 1 Open the Evaluation module and open a result Getting Started with KTA avant 29 0351 83 AA 137 8 Evaluate and print the results 8 3 Print the results Step Action 2 Click the Report icon Generate Report Format Global Chromatogram Global Chromatogram including logs Global Chromatogram including peak table Global Chromatogram including pH curve Global Full report Global Full report including pH curve 3 In the Generate Report dialog e Select any of the predefined formats from the list e Click the Preview button Preview Result A preview of the report displays in the Customize Report window 4 Are you satisfied with the layout e If Yes continue to step 5 e If No select File Exit to return to the Generate Report dialog and choose another format 138 Getting Started with AKTA avant 29 0351 83 AA 8 Evaluate and print the results 8 3 Print the results Step Action 5 In the Customize Report dialog click the Print icon Result The Print dialog opens Printer Microsoft Office Document Image Writer Y Properties Print Range Copies all ay Number of copies 1 O Pages Enter page numbers and or page ranges seperated by commas For example 1 3 5 23 6 In the Print dialog select Printer from the list and click OK Result Your report is printed Getting Started with AKTA avant 29 0351 83 AA 139
75. pH flow cell by injecting water into pH valve port Cal using a new syringe 8 Enter the pH of the second pH standard buffer in the pH for buffer 2 field 9 Repeat steps 5 6 using the second pH standard buffer Result The calibration date and time are displayed in the dialog and also values for Calibrated electrode slope and Asymmetry potential at pH 7 10 Is the Calibrated electrode slope 2 80 and the Asymmetry potential at pH 7 inside the interval 60 mV If Yes Click to switch the pH valve back to the default position and to close the Calibration dialog e If No Clean the pH electrode and repeat the calibration procedure If this does not help replace the electrode For information about cleaning and replacing the pH electrode see User Manual Chapter Maintenance CAUTION pH electrode Handle the pH electrode with care The glass tip may break and cause injury Getting Started with AKTA avant 29 0351 83 AA 95 6 Prepare the system for a run 6 6 Prepare the Fraction collector 6 6 Prepare the Fraction collector Introduction This section describes how to prepare the Fraction collector For information regarding the types of deep well plates tubes and Cassettes see AKTA avant User Manual WARNING Fraction collector Do not fractionate flammable liquids When running RPC methods or other procedures using organic solvents collect fractions through the Outlet valve Prepare the Fractio
76. re the system for a run 6 3 Prime buffer inlets and purge system pumps Step Action 3 Connect a 25 to 30 ml syringe to the purge valve of the left pump head of System pump B Make sure that the syringe fits tightly into the purge con nector Open the purge valve by turning it counter clockwise about 3 quarters of a turn Draw a small volume of liquid slowly into the syringe with a rate of about 1 ml per second Close the purge valve by turning it clockwise Disconnect the syringe and discard its contents Connect the syringe to the purge valve on the right pump head of System pump B and repeat steps 4 and 5 Keep the system flow running 86 Getting Started with AKTA avant 29 0351 83 AA 6 Prepare the system for a run 6 3 Prime buffer inlets and purge system pumps Purge System pump A Follow the instructions below to purge both pump heads of System pump A Step Action i 4 In the Manual instructions dialog e Select Pumps and pressures Gradient e Set Target to 0 B and Length to 0 min Click L Eeee _ Result Only System pump A is active In the Manual instructions dialog e Select Flow path Inlet A e Select the Position of one of the inlets to be used from the drop down list click LExecute_ Result Inlet valve A switches to the selected port Connect a 25 30 ml syringe to the purge valve of the left pump head of System pump A Make sure that the syringe fits tightly into
77. retical plate HETP and the peak asymme try factor AJ Intelligent Packing A flow of hydraulic liquid pushes the adapter down The user initiates the start of compression at the exact point when the adapter reaches the consolidated bed surface The adapter compresses the bed according to the packing factor or target bed height as selected Only available for AKTA avant 150 67 5 Create a method 5 2 Print a method 5 2 Printa method Introduction This section describes how to print the text instructions and variables of a method UNICORN uses the printers and printer settings that are installed on your computer How to print a method Follow the instruction below to print a method Step Action il Open the Method Editor module and click the Open Method Navigator icon in the toolbar a Result The Method Navigator pane opens Method Navigator Open amp Methods Method Q Folder name System A B Eric Methods Method1 System3 B Method2 System3 Results 2 Select the method to print and click the Open a Method icon in the navigator toolbar Open Result The selected method opens 68 Getting Started with AKTA avant 29 0351 83 AA 5 Create a method 5 2 Print a method Step Action 3 Click the Print icon Result The Print dialog opens Print Method 1 Printer MA x Properties Print items Phase range Text instructions All phases M V
78. rters of a turn Draw liquid slowly into the syringe until the liquid reaches the pump 5 Close the purge valve by turning it clockwise Disconnect the syringe and discard its contents 6 Repeat steps 2 5 for each piece of inlet tubing A that is to be used during the run Getting Started with AKTA avant 29 0351 83 AA 83 6 Prepare the system for a run 6 3 Prime buffer inlets and purge system pumps Prepare the system before purging the System pump heads Follow the instructions below to prepare the system 84 Step Action il Make sure that the piece of waste tubing connected to Injection valve port W1 is placed in a waste vessel 2 Open the System Control module and select Manual Execute Manual In structions Result The Manual instructions dialog opens 3 In the Manual instructions dialog e Select Flow path Injection valve and select System pump waste from the Position drop down list Click Execute Result The Injection valve switches to waste position This is necessary to achieve a low backpressure during the purge procedure e Select Pumps and pressures System flow For KTA avant 25 set the Flow rate to 1 0 ml min For AKTA avant 150 set the Flow rate to 10 0 ml min Click Execute Result A system flow starts Getting Started with AKTA avant 29 0351 83 AA 6 Prepare the system for a run 6 3 Prime buffer inlets and purge system pumps Purge System pump B Follow the instruction below to purg
79. ructions below e Keep UV flow cell clean Do not allow solutions containing dissolved salts proteins or other solid solutes to dry out in the flow cell Do not allow particles to enter the flow cell as damage to the flow cell may occur e Glass tube splinter Make sure to set the sample pressure be low the max pressure of the Superloop before executing a flow in the Manual instructions dialog when the Superloop is con nected e Avoid condensation If KTA avant is kept in a cold room cold cabinet or similar keep it switched on in order to avoid conden sation e Avoid overheating If KTA avant is kept in a cold cabinet and the cold cabinet is switched off make sure to switch off AKTA avant and keep the cold cabinet open to avoid overheating e Place the computer in room temperature If the KTA avant instrument is placed in a cold room use a cold room compatible computer or place the computer outside the cold room and use the Ethernet cable delivered with the instrument to connect to the computer 24 Getting Started with AKTA avant 29 0351 83 AA Maintenance 2 Safety instructions 2 1 Safety precautions WARNING To avoid personal injury when performing maintenance on the AKTA avant instrument follow the instructions below 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 do
80. s Meaning a ea o Warning 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 Supply voltage Make sure that the supply voltage at the wall outlet corresponds to the marking on the in strument before connecting the power cord Warning Fraction collector Do not fractionate flammable liq uids When running RPC methods or other procedures using organic solvents collect fractions through the Outlet valve The system complies with the requirements for elec tromagnetic compliance EMC in Australia and New Zealand The system complies with applicable European direc tives This symbol indicates that the system has been certi fied by a Nationally Recognized Testing Laboratory NRTL An NRTL is an organization that the Occupation al Safety and Health Administration OSHA has recog nized as meeting the legal requirements in USA title 29 of the Code of Federal Regulations 29 CFR Part 1910 7 Getting Started with AKTA avant 29 0351 83 AA 27 2 Safety instructions 2 2 Labels Labels concerning hazardous substances The following symbols on the labels concern hazardous substances This symbol indicates that electrical and electronic abel equipment must not be disposed of as unsorted municipal formation concerning the decommissioning o
81. s NO 700 am Pom ume 1000 sinn 000 25 000 we im ter v Corte te fw te weed comer oOw2 in Couma Wash ows i z 7 Use nanah pepsed tuben rane wai SF mo annati ate t guatr sson O Use aina tafa pepaaten BuiaPo Eneti ap serecs stam Area Description Phase Library Displays all available phases Getting Started with KTA avant 29 0351 83 AA 57 5 Create a method 5 1 Guide to method creation 5 1 1 Create and edit methods Area Description 2 Method Outline Presents an overview of the phases included in the active method 3 Phase Properties Presents detailed settings for the selected phase 4 Gradient Illustrates the gradient used in the active method Create a method Follow the instruction below to create and edit a chromatographic method based on a predefined method Step Action iL Open the Method Editor module and click the New Method icon Result The New Method dialog opens 2 Select System and Predefined Method in the dialog Click OK New Method Create a new method by using the Predefined Method Cation Exchange Chromatography CIEX O Empty Method Method Description A cation exchange column is equilibrated with start buffer eel is applied and positive charged proteins are bound Unbound material is washed away Proteins are eluted using a gradient of increasing concentration of e g NaCl Finally the column is washed with a high concentration of salt a
82. s how to start up the system both the software and the instrument It is assumed that your user profile is already created For information about how to set up and define users user groups and access rights please refer to UNICORN 6 Administration and Technical Manual Start the instrument and the computer 36 Follow the instructions below to start the instrument and the computer Step Action Ai Switch on the instrument by pressing the Power switch to the I position Result The instrument starts and the Instrument display states Not connect ed 2 Turn on the computer and monitor according to the manufacturer s instruc tions Getting Started with AKTA avant 29 0351 83 AA 3 Start up Start UNICORN and log on Follow the instruction below to start UNICORN and log on to the program Step Action 1 Double click the UNICORN icon on the desktop Result The Log On dialog opens Note If there is no connection to the database it is still possible to log on to UNICORN and control a running system The Log On dialog will give the option to start System Control without a database Click Start System Control to proceed to the next Log On dialog Getting Started with AKTA avant 29 0351 83 AA 37 3 Start up Step Action 2 In the Log On dialog e select User Name and e enter Password Note It is also possible to select the Use Windows Authentication checkbox and enter a network ID in th
83. s techniques see User Manual Stage Description Load The capillary loop is filled with sample Inject The sample is injected onto the column Getting Started with AKTA avant 29 0351 83 AA 7 Runa method 7 2 Applying the sample Sample application through a capillary loop A capillary loop is manually filled with sample using a syringe connected to the Injection valve port Syr During the method run the sample is automatically injected onto the column The loop is emptied and washed out using buffer from the system pumps The total buffer volume to be used for emptying and washing the capillary loop is set in the Phase Properties tab of the Sample Application phase at Empty loop with Phase Properties Text Instructions iT Sample Application V Use the same flow rate as in Method Settings Flowrate 10 000 ml min 0 000 25 000 Inject sample from loop Fill the loop using Manual load v Loop type Capillary loop v 6 00 mi O Inject sample directly onto column LS eS Sample inlet Fill loop 1 00 ml Empty loop with Sample volume U0 ml C Use the same inlets as in Method Settings Inlet amp Al v Inlet B B1 o0z 0 Fill the system with the selected buffer Tip Empty the capillary loop with a buffer volume that exceeds the volume of the loop This will ensure that the loop is completely emptied Getting Started with AKT
84. stem Calibrate Result The Calibration dialog opens Calibration System3 Monier to cate TT Curent vue Calibration procedure Prepare for calibration Calibration for pH electrode buffer 1 Description r pH calibrates the pH electrode with two point pH for buffer 1 L calibration i e using two pH calibration solutions It can also be used to fil the pH cell with storage solution since the pH valve is in calibration position pH for buffer 2 For calibration preferably use pH standard buffers pH 4 or 7 as the first calibration point and a pH standard buffer close to the lowest or highest pH Last calibrated on 2009 01 13 10 52 07 01 00 you need to measure as your second point Calibrated electrode slope Parameters pH for buffer 1 PH for buffer 2 Assymetiy potential at pH 7 mv 2 Set the pH monitor as the Monitor to calibrate by selecting pH from the list 3 Click Prepare for calibration i Result The pH valve switches to the calibration position 4 Enter the pH of the first pH standard buffer in the pH for buffer 1 field 5 Fill a syringe with approximately 10 ml of the first pH standard buffer Con nect the syringe to the Luer connector in pH valve port Cal and inject the buffer 94 Getting Started with KTA avant 29 0351 83 AA 6 Prepare the system for a run 6 5 Calibrate the pH monitor Step Action When the Current value is stable click cL 7 Wash the
85. sult The first New Column dialog opens New Column Column ID C The Column has a uniT ag has a fixed Code and exp Continue Cancel 112 Getting Started with AKTA avant 29 0351 83 AA 7 Runa method 7 3 Start a method run Step Action 7 Register the column using the 2D Barcode scanner as follows e Make sure that the pointer is placed in the first position of the Code field e Pointthe 2D Barcode scanner towards the data matrix tag on the column e Press and hold the trigger to create a beam e When the 2D Barcode scanner beeps the column ID is registered and displayed in the dialog T Maini e Alternatively manually enter the column ID that you find on the column label in the dialog using your keyboard e Click Continue Result The expanded New Column dialog opens New Column Column ID Alias optional Technique Desalting Column type HiPrep 26 10 Desalting C Use medium batch ID Getting Started with KTA avant 29 0351 83 AA 113 7 Runamethod 7 3 Start a method run Step Action 8 In the expanded New Column dialog e Enter Alias optional e Select Technique and Column type from the drop down lists e Check the Set medium expiration date check box and select a date from the drop down list e Click OK Tip Alias can be used for easy identification of a column Result The entered information is saved and the dialog closes 114 Getting Star
86. sult The selected phase is excluded from the method Rearrange phases within a method Follow the instruction below to rearrange phases within a method Step Action 1 Select the phase to move in the Method Outline pane Note The Method Settings phase cannot be moved 2 Drag and drop the phase to requested position in the Method Outline pane Result The selected phase is moved to the requested position Choose a maintenance method A number of predefined methods for preparation and cleaning are available Use these maintenance methods to prepare and clean the system and columns and to fill the system and columns with storage solution The table below gives suggestions for what methods and solutions to choose for the different purposes If you wish to Phase Method Solution Prepare tubing and instrument modules System Preparation suitable buffers before a run Prepare column before a run Column Preparation suitable buffers Getting Started with KTA avant 29 0351 83 AA 63 5 Create a method 5 1 Guide to method creation 5 1 1 Create and edit methods 64 If you wish to Phase Method Solution Clean tubing and modules System CIP 0 5 1 M NaOH Clean column Column CIP 0 5 1 M NaOH Leave system in storage solution System CIP 20 Ethanol Leave column in storage solution Column CIP 20 Ethanol Tip CIP Cleaning In Place and preparation of columns and system may be run eith
87. t 29 0351 83 AA 5 1 Introduction 1 1 Important user information 1 1 Important user information Read this before using the AKTA avant system All users must read the entire KTA avant Installation Guide and Getting Started with AKTA avant before installing operating or maintaining the instrument Do not operate the AKTA avant system 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 Purpose of Getting Started The purpose of this guide is to present a quick and easy guide to the system for a user with limited or no experience of UNICORN software and AKTA avant instrument The work flow is presented as practical instructions on how to operate the software and the instrument The instructions form a basic framework that you can expand on by reading selected parts in the other manuals This Getting Started guide includes the following topics e Basic features of KTA avant and UNICORN e Create methods e Prepare the system for runs e Perform runs e Make simple evaluations e Print reports For best results follow the guide from page to page in front of the system Prerequisites In order to follow this guide and use the system in the manner it is intended it is important that e you have a general understanding of how the computer and Windows work 6 Getting Started with AKTA avant 29
88. te the KTA avant instrument in any other way than described in the AKTA avant and UNICORN manuals e Operation and user maintenance of the KTA avant instrument should be performed by properly trained personnel only e To avoid exposing the column to excessive pressure make sure that the pressure limit is set to the specified maximum pressure of the column Before connecting a column to the AKTA avant instrument read the instructions for use of the column e Do not use any accessories not supplied or recommended by GE Healthcare e Do not use the KTA avant instrument if it is not working properly or if it has suffered any damage for example damage to the power cord or its plug damage caused by dropping the equipment damage caused by splashing liquid onto it NOTICE gt Avoid condensation by letting the instrument equilibrate to ambient temperature 16 Getting Started with KTA avant 29 0351 83 AA Using flammable liquids 2 Safety instructions 2 1 Safety precautions WARNING When using flammable liquids with the AKTA avant instrument follow these precautions to avoid any risk of fire or explosion Fire Hazard Before starting the system make sure that there are no unintentional leakage in the instrument or tubing Explosion hazard To avoid building up an explosive atmo sphere when using flammable liquids make sure that the room ventilation meets the local requirements Frac
89. ted with AKTA avant 29 0351 83 AA 7 Runa method 7 3 Start a method run Step Action 9 In the Select Columns dialog e Select Enter ID e Use the 2D Barcode scanner see step 7 to enter the column ID Select Columns System3 Enter or select a column individual Code Enter ID ke 2288 13 12345678 0000 00 a005 O Select ID 8 13 12345678 0000 00 0005 HiPrep 261 Disable column logging for this run Apply to all methods with the same column type Methods Remark Column Barcode Column type Column Handling Scouting run 1 0 0 Column Handling Scouting run 2 HiPrep 26 10 Desalting Column Handling Scouting run 3 HiPrep 26 10 Desalting Cancel e Alternatively choose Select ID and select the column individual to be used in the run from the drop down list Select Columns System3 Enter or select a column individual Code Enter ID Select ID Apply to all methods with the same column type Methods Remark Column Barcode Column Handling Column Handling Scouting run 2 HiPrep 26 10 Desalting Column Handling Scouting run 3 HiPrep 26 10 Desalting e Click OK Result The run starts All necessary actions occur automatically according to the method including ending of the run Getting Started with AKTA avant 29 0351 83 AA 115 7 Runamethod 7 4 Monitor the run 7 44 Monitor the run Introduction You may follow the on g
90. ting Started with AKTA avant 29 0351 83 AA 7 Runa method 7 4 Monitor the run e Click the Customize icon to open the Customize dialog tesa ug e Under the Run Log tab select the items to display Customize System3 Run Data Groups RunDataColor Curves Curve Style andColor XAxis YAxis Aunlog V Autoscroll Included entry types 7 Set Mark Block Start v V Alarms Warnings and Errors V Method vy v Manual System Included feedback types v v v v v Issued Processing Completed Failed Ignored Cancelled e Click OK to close the dialog Note These operations determine which data are shown in the Run Log pane during a run All original raw data are saved and can never be modified or removed from the result Flow scheme The Flow Scheme pane displays the current flow path during the run Color indication is applied as shown in the table below Real time data from monitors are also displayed in the flow scheme See illustration below Color Indication Green Open flow path Grey Closed flow path Red not shown Alarm Getting Started with AKTA avant 29 0351 83 AA 119 7 Runamethod 7 4 Monitor the run SyP to Col Syr to Loop to W Column PreC DeltaC UY 1 Cond pH Inlet B B 1 0 00 MPa 0 00 MPa 0 27 m
91. tion collector Do not fractionate flammable liquids When running RPC methods or other procedures using organic sol vents collect fractions through the Outlet valve RPC runs with 100 acetonitrile in AKTA avant 25 Always replace the green PEEK tubing between the used system pump and the pump pressure monitor with orange PEEK tubing i d 0 5 mm before running RPC with 100 acetonitrile Set the System pressure alarm to 10 MPa RPC runs with 100 acetonitrile in AKTA avant 150 Always replace the beige PEEK tubing between the used system pump and the pump pressure monitor with orange PEEK tubing i d 0 5 mm before running RPC with 100 acetonitrile Getting Started with AKTA avant 29 0351 83 AA 17 2 Safety instructions 2 1 Safety precautions Personal protection WARNING To avoid hazardous situations when working with the AKTA avant system take the following measures for personal protection e Always use appropriate personal protective equipment during operation and maintenance of AKTA avant system e Hazardous substances When using hazardous chemical and biological agents take all suitable protective measures such as wearing protective glasses and gloves resistant to the sub stances used Follow local and or national regulations for safe operation and maintenance of the system e Spread of biological agents The operator has to take all nec essary actions to avoid spreading hazardous biological agents
92. uency range 9 kHz to 25 MHz and inductive loop sys tems in the frequency range 9 kHz to 30 MHZ 301 489 3 Radio spectrum Matters ERM nized with EU directives ElectroMagnetic Compatibility EMC 1999 5 EC and standard for radio equipmentand 2004 108 EC services EN 300 330 2 Electromagnetic compatibility and EN standard is harmo nized with EU directive 1999 5 EC Getting Started with KTA avant 29 0351 83 AA 11 1 Introduction 1 2 Regulatory information FCC statement The AKTA avant instrument excluding transmitted radio frequency energy from the Mixer M9 and UV detector U9 D complies with FCC 47 CFR Part 15b Federal Communi cations Commission FCC title 47 of the Code of Federal Regulations CFR Part 15b Radio Frequency Devices Note This equipment has been tested and found to comply with the limits for a Class A digital device pursuant to part 15 of the FCC Rules These limits are designed to provide reasonable protection against harmful interference when the equipment is operated ina commercial environment This equipment generates uses and can radiate radio frequency energy and if not installed and used in accordance with the instruction manual may cause harmful interference to radio communications Operation of this equipment in a residential area is likely to cause harmful interference in which case the user will be required to correct the interference at his own expense The modules Mi
93. ult Anna 12 01 2008 14 12 lotta Anne 12201 2008 14 06 lotta Bjorn 01 14 2009 12 42 Default E Bimo 01 14 2009 12 44 Default Christer 12 01 2008 14 38 Default E Christine 12 02 2008 17 54 Default Claes 11 21 2008 08 32 Default B DefaultHome 09 25 2008 13 07 System E AsaDH 01 12 2009 15 56 Default lt Name UNTITLED System System3 o In the Save As dialog e Selecta target folder to enable the Save button e Type a Name for the method e Selecta System from the list e Click the Save button Save Result The created method is saved in the selected folder Add phases to a method Follow the instruction below to add phases to a method Step Action i 2 Select the desired phase in the Phase Library pane Drag and drop the phase to requested position in the Method Outline pane The phase can be placed at any postion below the Method Settings phase Result The phase is included in the method at the the requested position 62 Getting Started with AKTA avant 29 0351 83 AA 5 Create a method 5 1 Guide to method creation 5 1 1 Create and edit methods Delete phases from a method Follow the instruction below to delete phases from a method Step Action 1 Select the phase to delete in the Method Outline pane Note The Method Settings phase cannot be deleted 2 Click the Delete button located below the Method Outline pane Re
94. up flow downflow phases are automatically performed after the AxiChrom column has been packed Only available for AKTA avant 150 Reversed Phase Chromatography RPC After equilibration and sample application hydrophobic proteins adsorb to the column ligand After a wash to remove unbound sample elution is performed by generating a gradient of a non polar organic solvent such as Acetonitrile Finally the column is washed and re equilibrated System CIP The system is filled with cleaning solution Select for example inlets outlets and column positions to be cleaned Three System CIP phases are included in the method to facilitate the use of three different cleaning solution Additional System CIP phases can be added from the Phase Library if desired System Prepara tion The system is filled with preparation solution Select for example inlets outlets and column positions to be prepared Two System Preparation phases are included in the method Additional System Preparation phases can be added from the Phase Li brary if desired Getting Started with AKTA avant 29 0351 83 AA 5 1 3 Predefined phases 5 Create a method 5 1 Guide to method creation 5 1 3 Predefined phases The predefined phases that are available in the predefined methods and in the Phase Library are described in the table below Phase Description Method Settings The first and mandatory phase in any method Defines com
95. up to the time of the power failure is available in UNICORN 30 Getting Started with AKTA avant 29 0351 83 AA 2 Safety instructions 2 3 Emergency procedures Power failure to will result in Computer e The UNICORN computer shuts down e The instrument display shows status Not con nected e Therunis interrupted immediately e Data generated up to 10 seconds before the power failure can be recovered Note The UNICORN client may close down during a temporary overload of the processor This may appear as a computer failure The run continues and you can restart the UNICORN client to regain control Uninterruptible power supply UPS A UPS can prevent data loss during a power failure and allow time for a controlled shut down of AKTA avant For UPS power requirements see the system specifications in the AKTA avant User Manual Remember to also take into account the specifications for the computer and monitor Refer to the manufacturers documentation Getting Started with AKTA avant 29 0351 83 AA 31 2 Safety instructions 2 4 Recycling information 2 4 Recycling information Introduction This section describes the procedures for disposal and recycling of AKTA avant Decommissioning and disposal of the equipment When taking AKTA avant 25 and 150 out of service e The equipment must be decontaminated performed by an Administrator e The components must be separated a
96. utlet and ETFE o d Fingertight connector waste tubing 1 16 i d 1 0 1 16 mm AKTA avant 150 Scope ofuse Tubing Connector Photo Inlet tubing FEP o d 3 16 Tubing connector i d 2 9mm 5 16 Ferrule blue 3 16 Tubing tocon PEEK o d Fingertight connector nectcolumns 1 16 i d 1 0 1 16 mm Outlet tubing FEP o d 1 8 Tubing connector i d 1 6 mm 5 16 Ferrule yellow 1 8 Waste tubing ETFE o d Fingertight connector 1 16 i d 1 0 1 16 mm bag Getting Started with AKTA avant 29 0351 83 AA 75 6 Prepare the system for a run 6 2 Prepare the flow path Inlet ports The table below shows the inlet ports of Inlet valve A Inlet valve B and Sample inlet valve Valves and ports Illustrations Inlet valve A Inlet ports e A1 A7 Out V9 IA Inlet valve B Inlet ports e B1 B7 Out V9 IB Sample inlet valve Inlet ports e 1 S7 Buff Out V9 IS 76 Getting Started with AKTA avant 29 0351 83 AA 6 Prepare the system for a run 6 2 Prepare the flow path Prepare the inlet tubing Connect inlet tubing to the inlet ports that are to be used and immerse all inlet tubing that is to be used during the method run in the correct buffers Prepare the outlet tubing Connect outlet tubing to the outlet ports of Outlet valve that are to be used during the run If a fraction co
97. valuation module and click the Open Result Navigator icon en Result The Result Navigator opens Result Navigator Results RecentRuns Find Results Open oy Results DoE Results Folder name System a B Eric Methods B Results fo Runt System fad Runz System 2 Select the Results tab 3 Select the result to open and click the Open a Result button in the navigator toolbar Open Result The result is opened in a Chromatogram pane Available chro matograms and peak tables are displayed Getting Started with AKTA avant 29 0351 83 AA 8 Evaluate and print the results 8 1 View the results Illustration of the Chromatogram pane The Chromatogram pane displays the curves from the method run _ Chrom 1 UV 1_280_Chrom 1 id101b 001 Cone B_Chrom 4 id101b 001 Fraction_Chrom 1 id101b 001 UV 1_280 01 BASEM Edited_Chrom 1 Run 1 225 220 215 210 205 200 195 190 185 180 175 170 165 160 155 150 145 140 135 130 125 i 120 5 110 105 100 30 85 80 75 70 65 60 50 45 40 35 30 25 20 15 10 Obt Waste B10 Bat B12 cA C2 Out Waste Out Waste F354 mi 45 50 55 60 by o w a Y 8 P R 8 BR a Customize Chromatogram To change the chromatogram properties e Click the Customize icon to open the Customize dialog T uA d e Make appropriate selections under th
98. wer by pressing the handle upwards and pulling out the drawer Getting Started with AKTA avant 29 0351 83 AA 99 6 Prepare the system for a run 6 6 Prepare the Fraction collector Step Action 3 Place the 50 ml tube tray on the Tray support of the Frac drawer Make sure that the front of the tray marked with the GE logo faces the front of the drawer and is hooked onto the two pins Note The Cassette tray shall not be used when the 50 ml tube tray or the 250 ml bottle tray is placed in the Frac drawer 4 Close the Frac drawer Make sure that it snaps into closed position Cassette identification After the Frac drawer has been closed the Frac arm scans the Cassette type code of each Cassette to identify the Cassette types If deep well plates are used the instrument also identifies the types of deep well plates WARNING Moving parts in Fraction collector Do not open the Fraction col lector drawer when the Fraction collector is active If you need to access the Fraction collector press Pause and make sure that the movement has stopped before opening the drawer 100 Getting Started with KTA avant 29 0351 83 AA 6 Prepare the system for a run 6 6 Prepare the Fraction collector View Fraction collector content To view the content of the Fraction collector open the System control module and select View Fraction Collector Content Fraction Collector Content System3 3 3 3 Zz Q o 5
99. x operating pressure 20 MPa Wavelength 190 700 nm AKTA avant 150 Parameter Limits Flow rate Up to 150 ml min Note When running the Column packing flow instruction the maximum flow rate for AKTA avant 150 is 300 ml min Max operating pressure 5 MPa Getting Started with AKTA avant 29 0351 83 AA 41 4 The instrument and software 4 1 AKTA avant instrument overview Parameter Wavelength 190 700 nm Illustration of the main parts of the instrument The illustration below shows the location of the main parts of the instrument Part Function Part Function 1 Fraction collector 7 Holder rails 2 Buffer tray 8 Swivel foot Lock Unlock knob 3 Instrument display 9 Swivel foot 4 Wet side 10 Swing out toolbox 5 Foldable door 11 Power switch 6 Pump cover 42 Getting Started with KTA avant 29 0351 83 AA Illustration of the wet side modules of the instrument The illustration below shows the modules of the wet side of the instrument 1 234 5 4 The instrument and software 4 1 AKTA avant instrument overview Part Function Part Function 1 Injection valve 11 Inlet valve A 2 Flow restrictor 12 System pump A 3 pH valve 13 Mixer 4 Column valve 14 Pressure monitor of Sample pump 5 UV monitor 15 Sample pump 6 Conductivity monitor 16 Pump piston rinsing system tubes 7 Inlet valve B 17 Sample inlet valve 8 Quaternary valve 18 Outlet
100. xer M9 and UV detector U9 D comply with FCC 47 CFR Part 15c Note This device complies with Part 15c rules Operation is subject to the following two conditions e This device may not cause harmful interference and e This device must accept any interference received including interference that may cause undesired operation The user is cautioned that any changes or modifications not expressly approved by the manufacturer could void the user s authority to operate the equipment Software declaration of conformity UNICORN 6 is technically compatible with all relevant sections of FDA 21 CFR Part 11 A part 11 system assessment checklist is available on request through the local GEHC representative Regulatory compliance of connected equipment Any equipment connected to AKTA avant should meet the safety requirements of EN 61010 1 IEC 61010 1 or other relevant harmonized standards Within EU connected equipment must be CE marked 12 Getting Started with AKTA avant 29 0351 83 AA Environmental Conformity 1 Introduction 1 2 Regulatory information Regulation Title 2011 65 EU Restriction of Hazardous Substances ROHS Directive 2002 96 EC Waste Electrical and Electronic Equipment WEEE Direc tive Regulation EC No 1907 2006 Registration Evaluation Authorization and restriction of CHemicals REACH ACPEIP Getting Started with AKTA avant 29 0351 83 AA Administration
101. yourself Note Do not overtighten when connecting columns Overtightening might rupture the connectors or squeeze the tubing and thereby result in high back pressure Attach a column holder and connect a column Follow the instruction below to connect a column to the instrument Always use a column holder The column is connected to two opposite parts of the Column valve using appro priate tubing and connectors Step Action 1 Choose a column according to the column selections made in the method to be run Column selection Show by technique Anion Exchange x Column type HiScreen Capto Q a Column Properties Column volume 4 657 mi Pressure limit pre column 0 50 MPa 0 02 20 00 Pressure limit delta column 0 30 MPa 0 02 20 00 V Use flow restrictor Getting Started with AKTA avant 29 0351 83 AA 89 6 Prepare the system for a run 6 4 Connect a column Step Action 2 Attach an appropriate column holder to the rail on the instrument 3 Attach the column to the column holder 4 Connect a suitable tubing to a Column valve port for example port 2A if column position 2 was chosen in the method to be run 5 Open the System Control module and select Manual Execute Manual In structions Result The Manual instructions dialog opens 90 Getting Started with AKTA avant 29 0351 83 AA 6 Prepare the system for a run 6 4 Connect a column Step Action 6 In

Getting Started with ÄKTA avant

Contents

Download Pdf Manuals

Related Search

Related Contents

Getting Started with &Auml;KTA avant

Contents

Download Pdf Manuals

Related Search

Related Contents

Getting Started with ÄKTA avant