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ÄKTAavant and UNIC ORN 6

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1. Customize Run Data The Run Data pane shows real time data from available monitors during the run Run Data Groups Run Data Color Curves Curve Style To change what parameters to display in the pane M Rundata W System state M Acc volume ee M Block volume e Click the Customize icon to open the Ace tine lock time Customize dialog M Scouting no O System flow CO System pressure O Conc B C Sample flow CO Sample pressure A O Pret pressure a O DeltaC pressure O PostC pressure Ouv1 Ouve e Under the Run Data Groups tab Ouvs O pH select the parameters to display C BufferPro pH O Conc 01 X O Cone Q2 e Click OK to close the dialog O Cone Q3 O Conc Q4 O Cond 0 Cond Customize Chromatogram The Chromatogram pane displays a graphical presentation of registered data from available monitors during the run To change the chromatogram properties e Click the Customize icon to open the Customize dialog Getting Started with AKTAavant and UNICORN 6 28 9440 69 AB 107 8 Runa method 8 4 Monitor the run e Make appropriate selections under the tabs described in the table below 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 differ
2. e press the F1 key with the item of interest selected and in focus Note To find information about a phase make sure that the Phase Properties tab is selected before pressing the F1 key or e click the Help icon in the open dialog Getting Started with AKTAavant and UNICORN 6 28 9440 69 AB 37 4 The instrument and the software 4 2 UNICORN software overview If you want to then 38 navigate the online help select Help Help for in any of the UNICORN modules see illustration above in the TOC 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 access any of the manuals in PDF format select Help Help for in any of the UNICORN modules see illustration above in the TOC pane expand the heading UNICORN online documentation portal and select Online documentation 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 In the System Control module open a method select the instruction of interest in the Instruction box in the Text in
3. 7 Autoscroll Included entry types Included feedback types 7 Set Mark Issued Block Start Processing v 7 Alarms Wamings and Errors Completed V Method Failed v v Manual Ignored System 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 Getting Started with KTAavant and UNICORN 6 28 9440 69 AB 109 8 Runamethod 8 4 Monitor the run Flow scheme 110 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 SyP to Col Syr to Loop to W1_ Column Pret DeltaC Cond pH UV 1 Inlet B B 1 1 0 00 MPa 0 00 MPa 0 27 mAU 0 00 mS cm 0 00 Getting Started with AKTAavant and UNICORN 6 28 9440 69 AB 8 Runamethod 8 5 After run procedures 8 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 t
4. Do not use any accessories not supplied or recommended by GE Healthcare Do not use the AKTAavant 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 Getting Started with AKTAavant and UNICORN 6 28 9440 69 AB 13 2 Safety instructions 2 1 Safety precautions NOTICE Avoid condensation by letting the instrument equilibrate to ambient temperature Using flammable liquids WARNING When using flammable liquids with the AKTAavant 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 atmosphere when using flammable liquids make sure that the room ventilation meets the requirements specified in AKTAavant and UNICORN 6 Installation Guide Fraction collector Do not fractionate flammable liquids When running RPC methods or other procedures using solvent based buffers collect fractions through the outlet valve Personal protection Warnings WARNING To avoid hazardous situations when working with the AKTAavant system take the following measures for personal protection 14 Always use appropriate personal protective equip
5. Inlet ports 74 The table below shows what types of tubing and connectors to use for different purposes Scope ofuse Tubing Connector Photo Inlet tubing Teflon o d Tubing connector 1 8 1 8 i d 1 6 Ferrule yellow 1 8 E Tubing to PEEK o d Fingertight connector connect 1 16 i d 0 50 1 16 columns mm n Waste tubing Tefzel o d Fingertight connector 1 8 i d 1 0 1 16 mm 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 A5 S Out V9 IA Getting Started with KTAavant and UNICORN 6 28 9440 69 AB 7 Prepare the system for a run 7 2 Prepare the flow path Valves and p Illustrations Inlet valve B Inlet ports e B1 B7 Out V9 IB Sample inlet valve Inlet ports e 1 S7 Buff Out V9 IS XX Getting Started with AKTAavant and UNICORN 6 28 9440 69 AB 75 7 Prepare the system for a run 7 2 Prepare the flow path Waste ports 76 Valves and ports Injection valve Waste ports e W1 W2 The table below shows the waste ports of Injection valve pH valve and Outlet valve Illustrations pH valve Waste port e W3 Outlet valve Waste port e W Out6 v9 0 Getting Started with
6. 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 chromatograms and peak tables are displayed 116 Getting Started with AKTAavant and UNICORN 6 28 9440 69 AB 9 Evaluate and print the results 9 1 View the results Illustration of the Chromatogram pane The Chromatogram pane displays the curves from the method run _ Chrom t UV 1_280_Chrom 1 id101b 001 Cone B_Chrom 1 id101b 001 Fraction _Chrom 1 id101b 001 UV 1_280 01 BASEM Edited_Chrom 1 Run 1 Customize Chromatogram To change the chromatogram properties e Click the Customize icon to open the Customize dialog fom uA d e Make appropriate selections under the tabs described in the table below Getting Started with KTAavant and UNICORN 6 28 9440 69 AB 117 9 Evaluate and print the results 9 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
7. Getting Started with KTAavant and UNICORN 6 28 9440 69 AB 33 4 The instrument and the software 4 1 AKTAavant instrument overview Illustration of the wet side modules of the instrument The illustration below shows the modules of the wet side of the instrument 4 13 12 H Part Description 1 Injection valve 2 Column valve 3 Flow restrictor 4 pH valve 5 UV monitor 6 Conductivity monitor 7 Inlet valve B 8 Quaternary valve 9 System pump B 10 Pressure monitor of System pumps 11 Inlet valve A 34 Getting Started with KTAavant and UNICORN 6 28 9440 69 AB 4 The instrument and the software 4 1 AKTAavant instrument overview Part Description 12 System pump A 13 Mixer 14 Pressure monitor of Sample pump 15 Sample pump 16 Pump piston rinsing system tubes 17 Sample inlet valve 18 Outlet valve 19 Holder rails Getting Started with KTAavant and UNICORN 6 28 9440 69 AB 35 4 The instrument and the 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
8. Getting Started with AKTAavant and UNICORN 6 28 9440 69 AB 2 Safety instructions 2 1 Safety precautions Maintenance Warnings WARNING To avoid personal injury when performing maintenance on the AKTAavant instrument follow the instructions below e 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 e Disconnect power Always switch off power to the instrument before replacing any component on the instrument unless stated otherwise in the user documentation e 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 NOTICE Cleaning Keep the instrument dry and clean Wipe regularly with a soft damp tissue and if necessary a mild cleaning agent Let the instrument dry completely before use Getting Started with KTAavant and UNICORN 6 28 9440 69 AB 19 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 AKTAavant instrument For information about marking of the computer equipment refer to the manufacturer s instructions Labels on the AKTAavant instrument The illustrations below show the labels that are
9. KTAavant and UNICORN 6 28 9440 69 AB 7 Prepare the system for a run 7 2 Prepare the flow path Prepare inlets and outlets Follow the instruction below to prepare the inlets and outlets that are to be used in the method run Step Action 1 Connect inlet and outlet tubing to the ports that are to be used 2 Immerse all inlet tubing that is to be used during the method run in the correct buffers 3 Immerse the waste tubing from the Injection valve the pH valve the Outlet valve and the Fraction collector into a sufficiently large waste vessel typically 2 10 liters Getting Started with AKTAavant and UNICORN 6 28 9440 69 AB 77 7 Prepare the system for a run 7 3 Prime buffer inlets and purge System pumps 7 3 Prime buffer inlets and purge System pumps Introduction Before usage of the System pumps it is important to 1 Prime the inlets fill the buffer inlets with liquid 2 Purge the System pumps remove air from the pumps Prime buffer inlets Follow the instruction below to prime the buffer inlets Each buffer inlet that is to be used during the method run has to be primed Step Action 1 Make sure that all buffer inlet tubing that is to be used during the method run is immersed in the correct buffers 2 Make sure that the waste tube connected to Injection valve port W1 is immersed in a waste vessel 3 Open the System Control module in UNICORN and select Manual Execute Manual Instructions Re
10. Step Action 1 Switch on the instrument by pressing the Power switch to the I position 2 Open the the System Control module in UNICORN and click the Connect to Systems icon Connect to Systems Connected systems 1 selected max 3 System name Control View O E System OE System E System3 O eH System4 E g Systems O amp Systeme Op System Getting Started with KTAavant and UNICORN 6 28 9440 69 AB 29 30 3 Start up 3 1 Start the system and log on to UNICORN Step Action 3 In the Connect to Systems dialog e Select the instrument e Select Control mode e Click OK Result The selected instrument can now be controlled by the software Getting Started with AKTAavant and UNICORN 6 28 9440 69 AB 4 The instrument and the software 4 The instrument and the software Introduction This chapter gives an overview of the AKTAavant system instrument software and accessories Illustration of the system The illustration below shows the AKTAavant instrument with UNICORN software installed on a computer Contents This chapter contains the following sections Section See page 4 1 AKTAavant instrument overview 32 4 2 UNICORN software overview 36 4 3 UNICORN software modules 39 Getting Started with AKTAavant and UNICORN 6 28 9440 69 AB 31 4 The instrument and the software 4 1 AkKTAavant instrument overview 4 1 AKTAavant instrument overview Introduction This secti
11. Z Auto update parameter fields will be updated during method run 5 e Click L Erscute _ Click L Execute Result The Column valve switches to position 2 Getting Started with AKTAavant and UNICORN 6 28 9440 69 AB 85 7 Prepare the system for a run 7 4 Connect a column Step Action 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 Pumps and pressures System flow Sample flow Gradient Pump wash System wash Off Wash settings BufferPro pH Pressure control parameters How path t Monitors H Fraction collection Parameters for System flow amp Flow rate 0 000 25 000 0 2 ml min Pressure control Save result as Auto update parameter fields will be updated during method run Click E Result A system flow of 0 2 ml min starts 8 When buffer leaves the tube on port 2A in a continuous mode and the top part of the column is filled with buffer connect the tube to the top of the column 86 Getting Started with AKTAavant and UNICORN 6 28 9440 69 AB 7 Prepare the system for a run 7 4 Connect a column Step Action 9 Connect a tube to the bottom of the column 10 When buffer leaves the tube at the bottom of the column in a continuous mode connect this tube to the Column valve Use the port opposite to the one already connected to the column in this
12. window 126 Getting Started with AKTAavant and UNICORN 6 28 9440 69 AB 9 Evaluate and print the results 9 3 Print the results Step Action 4 Are you satisfied with the layout e If Yes continue to step 5 e If No select File Exit to return to the Print Chromatograms dialog and choose another print format 5 In the Customize Report window click the Print icon Result The Print dialog opens Printer Microsoft Office Document Image Whiter im Properties Print Range Copies all Pages Enter page numbers and or page ranges seperated by commas For example 1 3 5 23 Number of copies ji j 6 In the Print dialog select Printer from the list and click OK Result Your chromatogram is printed Getting Started with KTAavant and UNICORN 6 28 9440 69 AB 127 9 Evaluate and print the results 9 3 Print the results Print a report Follow the instruction below to print a standard format report Step Action i Open the Evaluation module and open a result 2 Click the Report icon m Result The Generate Report dialog opens 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 Prev
13. 7 4 Connect a COlUMN ae eeesessessssssssssssssssssssssssssssnsesssnsesssssuanssnussssanunssinusssntsunsnsnsusitsnsssnssnuetseneeseeees 83 7 5 Calibrate the pH MOnItOr wo ececsscsssssssssssseessssescccssssssssussssssessccsssssssssssusssessccssssssssusssssesssessesesssnssnees 88 7 6 Prepare the Fraction Colector o eccecssssssssssssssssescssesssssssssssssscsssssssssssssssecsesessssssssssnusessessecesssesssee 90 amp R mamethod Ran ee Oe Onn erst fH ere te ener neg ere pln aaa eaaa en erate 95 Bl Before OUR SEGUE ryser inen Naena Sta EE Ea ESEA S Bonsarsnensaisiasaielootitesnseandenedunyetiareern sate 96 82 APplyingihe samples eigin 98 83 Statt GIMENO UN ienssnieeiinaiann nanan anna n aa 101 Getting Started with KTAavant and UNICORN 6 28 9440 69 AB 3 Table of Contents BAe MOIR tS PUI sessatashass deccovssssssastusnichsnttectssussetnloastea hvaass sss anaa Anii 106 8 5 After run procedures 111 9 Evaluate ana Print the results rsrsrsrsrs 115 gE MIS AINE results lt sssssstiasssacccnsssasszataanhcbsnctactunsscoasecansenashssasvea an Anaan aii 116 9 2 PeakintegratiON sassidissssdescesazsssisbaacssavectussstevasabevsdea Avassvea ssn i a RERA 120 93 Prnttheresutsceinpsianma a aa aa A NEN NN 125 NO N eRe E A E EE E E E E T E T EE 130 10 1 M intenance POJAM eeasnnmnsia nN AAN E EE RE 131 4 Getting Started with KTAavant and UNICORN 6 28 9440 69 AB 1 About this guide 1 About this guide Introduction This chapter describes the pur
14. 8 Runa method 8 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 instruction below to fill the pH flow cell with storage solution The calibration function is used to switch the pH valve However no calibration is performed 112 Step Action i Open the System Control module and select System Calibrate Result The Calibration dialog opens Calibration System3 Monitor to calibrate iE Curent value Calibration procedure Calibration for pH electrode buffer 1 Description pH for butler isaten to unig to PE maitin mado 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 butters 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 fet foi pad ih a TAN J D ee y N 2 In the Calibration dialog select pH from the Monitor to calibrate drop d
15. 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 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 button and select Reset Zoom Result The original area is restored in one step 118 Getting Started with AKTAavant and UNICORN 6 28 9440 69 AB 9 Evaluate and print the results 9 1 View the results View documentation To view documentation of the run Click the View Documentation icon to open the Documentation dialog Select the tab of interest Which tabs are displayed depends on the settings in the active method Click OK to close the dialog Documentation Fraction Collector Variable List Scouting Textinstructions Notes BufferPta_ Columns Evaluation Procedures Fun Log Method Information nation Star
16. 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 completely filled Excess of sample will leave the valve through port W1 100 Getting Started with AKTAavant and UNICORN 6 28 9440 69 AB 8 Runa method 8 3 Start a method run 8 3 Start a method 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 im Result The Method Navigator pane opens Method Navigator Open amp Methods Method Q Folder name System A 2 Eric E Methods al Method1 System3 Ell Method2 System3 F Results Select the method to run and click the Run icon aza el 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 AKTAavant a
17. 100 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 e Moving the instrument horizontally Three persons are recommended when moving the instrument horizontally e Supply voltage Make sure that the supply voltage at the wall outlet corresponds to the marking on the instrument before connecting the power cord e Protective ground The AKTAavant instrument must always be connected to a grounded power outlet e Power cord Only use power cords delivered or approved by GE Healthcare Getting Started with AKTAavant and UNICORN 6 28 9440 69 AB 15 2 Safety instructions 2 1 Safety precautions 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 manufacturer of the computer Notices NOTICE To avoid damage to KTAavant or other equipment when installing or moving the instrument follow the instructions below Vents on the KTAavant instrument To ensure adequate ventilation keep papers and other objects away from the vents of the instrument Disconnect power To prevent equipment damage always switch off power to the AKTAavant instrument before an ins
18. Select System and Predefined Method in the dialog Click OK a Create a new method by uring the Predetined Method Canon Exchange Cheomatogagty O09 O Empty Method Meiteni Descagten A caon comnis equilbroted with wart butter C caveat Result The phases included in the chosen method show in the Method Outline pane and the default settings for each of the phases show in the Phase Properties pane Getting Started with KTAavant and UNICORN 6 28 9440 69 AB Step Action 6 Create a method 6 1 Guide to method creation 6 1 1 Create and edit methods 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 Whana Progar Test irmtracters T Mothod Setinge Cohan setectons Show by technique Cation Exchange Commie ary Cohen vine 0100 si Sampie Application Poecuze k precoken 2000 MPa J902 20 00 gt Presnse int detacotmn 2000 MPa P02 2000 EE Ure thaw sasinen Corean Wash Cohen pesten Bppar v Ernon Fom isie 1 000 mimi 49 000 26 000 X Z Connal he fos to avoid overprorsue Conan Wash v Ute narasi preporend badors neta ay bee by Equiibration O Ute Dutta ro moetan tader p
19. 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 A Administration WF Evaluation mS Control ig 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 36 Getting Started with AKTAavant and UNICORN 6 28 9440 69 AB Column Handling Ctrl L 4 The instrument and the software 4 2 UNICORN software overview Administration Method Editor System Control Options Change Passwords Access the help utility A comprehensive help utility is included in the UNICORN software The table below describes how to access the different parts of the help utility If you want to then find information about a UNICORN module select Help Help for in the UNICORN module of interest Contextual Help Fi Instruction Set About UNICORN find information about the item currently selected and in focus e g a pane a dialog or a method phase
20. clicking the pin symbol in the top right hand corner of the pane illustrated below If Auto Hide is selected the panes open automatically when the mouse pointer is placed over the corresponding tab and remain open as long as the mouse pointer remains over the pane The pane closes automatically when the pointer is moved outside op Getting Started with KTAavant and UNICORN 6 28 9440 69 AB 5 Files and folders in UNICORN 5 1 Navigator overview Navigator toolbar icons The table below shows the common navigator toolbar icons Icon Function Open Open Opens the selected file Pa New folder Creates a new folder in the folder that is currently selected G Refresh Updates all items in the navigator pane to the current status Opens the View Details droplist see illustration below where the following optional information may be selected for display in the navigator pane e System e Created by user name of the person who created the original file e Last modified date and time when then file or folder was last modified v System v Createdby v Last modified Getting Started with KTAavant and UNICORN 6 28 9440 69 AB 49 5 Files and folders in UNICORN 5 1 Navigator overview Filter settings By default the Method Navigator shows only methods and the Result Navigator only results This is the Default Filter setting for the items that are displayed However each navi
21. is selected in the navigator pane The search can be continued step by step until the whole folder structure has been searched 54 Getting Started with AKTAavant and UNICORN 6 28 9440 69 AB 6 Create a method 6 Create a method Introduction 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 Contents This chapter contains the following sections Section See page 6 1 Guide to method creation 56 6 2 Print a method 68 Getting Started with AKTAavant and UNICORN 6 28 9440 69 AB 55 6 Create a method 6 1 Guide to method creation 6 1 Guide to method creation Introduction 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 You can create both purification methods and maintenance methods You can also add a maintenance phase at the end of a purification method For further information about method creation see UNICORN 6 Method Manual Contents This sections contains the following subsections Section See page 6 1 1 Create and edit methods 57 6 1 2 Predefined methods 65 6 1 3 Predefined phases 67 56 Getting Started with AKTAavant and UNICORN 6 28 9440 69 AB 6 Create a method 6 1 Guide to method creation 6 1 1 Create and edit methods 6 1 1 Create and edit
22. 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 shows the Method Editor module and its panes Getting Started with AKTAavant and UNICORN 6 28 9440 69 AB 57 6 Create a method 6 1 Guide to method creation 6 1 1 Create and edit methods Part Description Create a method 58 1 Phase Library Displays all available phases 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 Follow the instruction below to create and edit a chromatographic method based on a predefined method Step Action nh Open the Method Editor module and click the New Method icon Result The New Method dialog opens 2
23. placed first in alphabetical order and e individual files are placed in alphabetical order after the folders Note The sorting order may be changed by clicking the header of each column You can also move folders and files by using Cut and Paste in the same manner as Copy and Paste 52 Getting Started with AKTAavant and UNICORN 6 28 9440 69 AB 5 Files and folders in UNICORN 5 2 Handling files and folders Delete files and folders Follow the instruction below to delete files and folders in the Method Navigator or in the Result Navigator Note Step Top level folders for example home folders cannot be deleted this way Refer to UNICORN 6 Administration and Technical Manual for more information about home folders Action Select the item s that you want to delete Choose Edit Delete Result A dialog for confirmation of the delete action opens Press Yes to confirm the delete action in the confirmation dialog Result The selected items are deleted Note If you have chosen to delete a folder that is not empty a special confirmation dialog opens This dialog displays all the items that will be deleted as a consequence both items in the active UNICORN module and items in other UNICORN modules see illustration below Delete Confirmation The folder s you are about to delete is not empty Continue anyway amp Folders 1 Al Methods 9 fq Results 8 Lo Queues 0 Rename files and fold
24. 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 Getting Started with AKTAavant and UNICORN 6 28 9440 69 AB 91 7 Prepare the system for a run 7 6 Prepare the Fraction collector 92 Step Action 4 5 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 Getting Started with AKTAavant and UNICORN 6 28 9440 69 AB 7 Prepare the system for a run 7 6 Prepare the Fraction collector Step Action 6 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 7 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 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 sto
25. u Pause Suspends the method run and stops all pumps Continue Resumes for example a held or paused End Permanently ends the method run groups and run log contents can be set I gt Lal method run m ad Customize Opens the Connect to Systems Opens Ay Customize dialog where P the Connect to Systems A curve settings run data KA dialog where systems can be connected and currently connected users are displayed 44 Getting Started with KTAavant and UNICORN 6 28 9440 69 AB 4 The instrument and the 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 chromatography 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 feats Recent Pun Fret Rents on fp EY ents cotra E Fedde name EJ UNCOPN Database Buwa Dira nat SA 882 388828 E no 100 n Peak name Nerini Ase let weil Wea AU n9 W eS ne samas 10849
26. 2 Safety instructions 2 1 Safety precautions 18 Cautions CAUTION To avoid hazardous situations when operating the AKTAavant system follow the instructions below Risk of breaking test vials Do not use excessive force to press vials with incorrect dimensions into the Fraction collector cassettes Glass vials may break and cause injuries 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 pH electrode Handle the pH electrode with care The glass tip may break and cause injury Max weight on top tray Do not place containers with a volume of more than 10 liters on the top tray The total allowed weight on the top tray is 40 kg Max size of bottles on front panel Do not fasten bottles with a volume of more than 1 liter in the front panel rails Notices NOTICE To avoid damage to AKTAavant or other equipment when operating the instrument follow the instructions below 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 Waste container height The maximum level of the waste container content must be lower than 30 cm above the lab bench for trouble free use
27. 69 AB 3 Start up 3 1 Start the system and log on to UNICORN 3 1 Start the system and log on to UNICORN Introduction This section describes how to start UNICORN and the instrument and how to connect to the system Start UNICORN and log on Follow the instruction below to start and log on to UNICORN Step Action 1 To start the program double click the UNICORN icon on the desktop fa UNICORN 6 0 Result The Log On dialog opens ff Log On UNICORN User Name Default Password Getting Started with KTAavant and UNICORN 6 28 9440 69 AB 27 3 Start up 3 1 Start the system and log on to UNICORN Step Action 2 In the Log On dialog e Select your User Name from the drop down list e Enter your Password e Click the Options button Result Checkboxes for each of the UNICORN modules are displayed in the Log On dialog f Log On UNICORN User Name Eric Password Start UNICORN Administration V System Control Method Editor Evaluation 3 In the Log On dialog e Select which UNICORN modules to start by clicking the corresponding checkboxes e Click OK Result The selected UNICORN modules open 28 Getting Started with KTAavant and UNICORN 6 28 9440 69 AB 3 Start up 3 1 Start the system and log on to UNICORN Start the instrument and connect to the system Follow the instruction below to start the instrument and connect to the UNICORN software
28. 69 AB 7 1 About this guide 1 1 Get started Intended use AkTAavant is a liquid chromatography system intended for method development The system can be used to screen for optimal choice of columns media and running parameters to purify selected proteins The AKTAavant 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 proceed 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 Getting Started with KTAavant and UNICORN 6 28 9440 69 AB 1 About this guide 1 1 Get started Typographical conventions Software items are identified in the text by bold italic text A colon separates menu levels thus File Open refers to the Open command in the File menu Hardware items are identified in th
29. 7 EEJ eur Getting Started with AKTAavant and UNICORN 6 28 9440 69 AB 45 4 The instrument and the 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 guide 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 chromatogram can be printed Copy Copies the selected result or folder Paste Pastes a copied result or folder to the selected folder E E Undo Restores the method to the state it was in before the last change Report Opens the Create report dialog where a report 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 Customize 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 integrated 46 Getting Started with KTAavant and UNICORN 6 28 9440 69 AB 5 Files and folders in UNICORN 5 Files and folders in UNICORN Introduction Contents In the UNICORN user interface method and result data are represented by f
30. Cond temp_Chrom 1 Manual Run 001 013 Sample flow_Chrom 1 Manual Run 001 K 014 Sample pressure_Chrom 1 Manual Run 001 015 PreC pressure_Chrom 1 Manual Run 001 iM 016 DeltaC pressure_Chrom 1 Manual Run 001 020 PostC pressure_Chrom 1 Manual Run 001 027 Cone 1_Chrom 1 Manual Run 001 y UV 1_280 01 PEAK Peak Window Accept negative peak Peak table name Basain Column height 0 00 cm Column Vt Save and Edit Peak Table OK Cancel 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 AKTAavant and UNICORN 6 28 9440 69 AB 121 9 Evaluate and print the results 9 2 Peak integration Illustration of Chromatogram and Peak Table 122 The illustration below shows a Chromatogram pane including a Peak Table 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 10 5 0 5 10 15 20 25 30 35 40 45 50 55 60 A UV 1_280 01 PEAK Edited Peak data Integration summary No Peak name Retention ml Area ml mAU Height mAU LE 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 Getting Started with KTAavant and UNICORN 6 28 9440 69 AB 9 Evaluate and print the results 9 2 Pea
31. GE Healthcare Getting Started with AKTAavant and UNICORN 6 KTA Table of Contents Table of Contents 1 About this guide 5 1 1 Get Started wees 6 1 2 Regulatory INFOMATION sssssessssssssscsssssssssssssessessccsssssssssussssesecsssssssssussusssesseesessesssusssessesssessssenssessiees 10 2 Safety e gs Co p ene Rn eevee et Reet see tesi aE a PUP NTU eiaeoei mete 12 21 Safety FESCUE OMS sssescssstddssossssavasscetbclancsssvvssisasoososddeods suutesvisssssostacccedssseiiashsstbadannsnnnsdesssttiatettocvntaien 13 2 2 BOIS csseccsscesesassestesosvscasoscatszsstevasassvnvasvecesseestezs Sivevois A T AAAG 20 2 53 Emergenty proc dures nsssssnianmnii uniana A aR 23 E o A E AE E E E ET E TT EET ET 26 3 1 Start the system and log on to UNICORN ssssssssssessssscccsssssssssssssssseccssssssssunsessssssesssssssssssseesees 27 4 The instrument and the software eeesesessesoseossesossesossssescossesesessesseseseosseseses 31 4 1 KTAavant instrument OVEFVIEW ccssssssssssssssssssssessssssessssssrsssssssssstersstterssstessstteststttesesttsestttedsetteeees 32 4 2 UNICORN software Overview iececccsescssssessssssessssssessssssessesssesessssesssuusesssuesessssusessssesessssesesssseessssueess 36 4 3 UNICORN SoftWare MOdUlES na enar E RAAR 39 43 1 Method Editor mod le siscisescstsscciscacassstdecscscatitessatevasvznassdiovssnsccdeseanadicsatansasonsseedigua nasctiigneniocit 40 432 System Control MOUIE ssssssssss
32. In the Manual instructions dialog e Select Pumps and pressures Pump wash and select one inlet from the Inlet A or Inlet B drop down list Preferably choose the buffer inlet to be used in the beginning of the method run Manual instructions System3 Instructions nae i arameters for Pump was Pumps and pressures A Inlet A System flow i Sample flow ee Gradient Inlet B m Pump wash i m System wash OFF j 4 Wash settings BufferPro Q inlets BufferPro pH r 1 L Pressure control parameters Or vie S Aow path Sample inlet 4 Monitors ies Fraction collection x or ja Rat 1 Save result as Browse 2 vV Auto update parameter fields will be updated during method run 3 e Click L Erscute _ cick E Result A pump wash of the selected inlet starts Getting Started with KTAavant and UNICORN 6 28 9440 69 AB 81 7 Prepare the system for a run 7 3 Prime buffer inlets and purge System pumps Step Action 2 Connect a 25 30 ml syringe to the left purge valve of the selected system pump Make sure that the syringe fits tightly into the purge connector 3 Open the purge valve by turning it counterclockwise about 3 4 of a turn Draw 5 10 ml of liquid slowly into the syringe with a rate of about 1 ml s 4 Close the purge valve by turning it clockwise Disconnect the syringe and discard its contents 5 Connect the syringe to the purge valve on the right pump head of t
33. Navigator Step Action al Select the folder in which you want to create the new sub folder 2 Click the New Folder icon t Result A sub folder named New Folder is created in the selected folder Jj Methods Du New Folder Note The new folder is placed after all other sub folders but before any files directly in the selected folder 3 Type a name for the new folder Result The new folder is renamed Getting Started with AKTAavant and UNICORN 6 28 9440 69 AB 51 5 Files and folders in UNICORN 5 2 Handling files and folders Copy and paste files and folders Follow the instruction below to copy and paste files and folders in the Method Navigator or in the Result Navigator If you copy a folder you will also copy all the files and folders that it contains Step Action 1 Select one or several files and or folders 2 Click the Copy toolbar icon O Result The selected items are copied 3 Select a target folder 4 Click the Paste toolbar icon P m Result The copied items are pasted into the selected folder TIP You can select a range of items by pressing the keyboard Shift key or several individual items by pressing the keyboard Ctrl key Move files and folders Files and folders may be moved by drag and drop within the folder structure that you have access to The destination must always be a folder and the contents will normally be re sorted automatically so that e folders are
34. P 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 either as separate predefined methods or as phases included in chromatographic 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 KTAavant and UNICORN 6 28 9440 69 AB 6 1 2 Predefined methods 6 Create a method 6 1 Guide to method creation 6 1 2 Predefined methods The predefined methods that are available are described in the table below Method Description Affinity chromatography AC After equilibration and sample application the protein of interest is adsorbed to the column ligand After a wash to remove unbound 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 s
35. ant and UNICORN 6 28 9440 69 AB Step 6 Create a method 6 1 Guide to method creation 6 1 1 Create and edit methods Action In the Phase Properties pane of the Method Settings phase click the Start Protocol button if you wish to include a start protocol Start Protocol Result The Start Protocol dialog opens Start Protocol Method Items to display at methad start Fraction Collection Current setting item description Variable List Scouting Displays the fraction collector setup tube type and position Text Instructions Notes Gradient BufferPro Columns Evaluation Procedures Method Information System Settings Calibration Questions Changeable Batch ID V Result Name and Location Deine Dues 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 Select the next phase in the Method Outline pane by clicking it and choose appropriate parameter values in the Phase Properties pane 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 AKTAavant and UNICORN 6 28 9440 69 AB 61 6 Create a method 6 1 Guide to method creatio
36. are the system 71 7 2 Prepare the flow path 72 7 3 Prime buffer inlets and purge System pumps 78 7 4 Connect a column 83 7 5 Calibrate the pH monitor 88 7 6 Prepare the Fraction collector 90 70 Getting Started with AKTAavant and UNICORN 6 28 9440 69 AB 7 Prepare the system for a run 7 1 Before you prepare the system 7 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 which valve ports to use for inlets and outlets which cassettes with corresponding deep well plates and or tubes to use in the Fraction collector if applicable which column type to use which column position to use which sample application technique to use that the pH electrode is connected if applicable if it is a reversed phase chromatography RPC run WARNING RPC runs with 100 acetonitrile Always replace the green PEEK tubing between the used system pump and the 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 WARNING Fraction collector Do not fractionate flammable liquids When running RPC methods or ot
37. ation module The efficiency of the column is determined in terms of height equivalent to a theoretical plate HETP and the peak asymmetry factor A3 The result is logged in the column logbook Column Preparation 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 AKTAavant and UNICORN 6 28 9440 69 AB 65 6 Create a method 6 1 Guide to method creation 6 1 2 Predefined methods Method Description 66 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 Interaction Chromatography HIC After equilibration and sample application use a buffer containing a high salt concentration for example 2 M Ammonium Sulphate 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 Reversed Phase Chromatography RPC After equilibration and sample application hydrophobic proteins adsorb to the column ligand After a wash to r
38. attached to the AKTAavant instrument resye 4r Oitavo Code Me IODD Supply Velloge 120 240 V sotonu Power Comumgten 800 Paises Ge WPL wet de B22 AZ CCC mame Saris aaas a 20 Getting Started with KTAavant and UNICORN 6 28 9440 69 AB Safety symbols 2 Safety instructions 2 2 Labels The following safety symbols are used in the labels Label Meaning 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 instrument before connecting the power cord Warning Fraction collector Do not fractionate flammable liquids When running RPC methods or other procedures using solvent based buffers collect fractions through the outlet valve The system complies with the requirements for electromagnetic compliance EMC in Australia and New Zealand The system complies with applicable European directives Getting Started with KTAavant and UNICORN 6 28 9440 69 AB 21 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
39. avant and UNICORN 6 28 9440 69 AB 123 9 Evaluate and print the results 9 2 Peak integration 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 Save results Peak Retention Area of total of total Height name ml ml mAU area peak area m4U i 31 93 206 2683 26 47 47 39 164 965 2 33 58 94 1728 12 08 21 64 188 497 3 I 33 84 121 6052 15 60 27 94 225 321 4 34 78 13 1725 1 69 3 03 6 747 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 id save the changed result as a new result 124 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 AKTAavant and UNICORN 6 28 9440 69 AB 9 Evaluate and print the results 9 3 Print the results 9 3 Print the res
40. buffer inlet tubing is immersed in correct buffer vessels consider solution identity and volume the waste tubing is immersed into appropriate waste vessels consider vessel size and vessel material no tubing is twisted and the flow path is free from leakage Hold pause or stop the run At the end of a method the run stops automatically All pumps stop and the system makes an acoustic end signal and displays End in the Run Log 96 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 ich temporarily pause the method and stop click the Pause icon all pumps tbl Getting Started with AKTAavant and UNICORN 6 28 9440 69 AB 8 Runamethod 8 1 Before you start If you want to resume for example a held or paused click the Continue icon method run I gt Note An ended method cannot be continued permanently end the run click the End icon ey a Note When ending a method run in advance it is possible to save the partial result Warnings concerning use of hazardous substances WARNING Hazardous chemicals during run When using hazardous chemicals run System CIP and Column CIP to
41. ccording to the column selections made in the method to be run Column selection Show by technique Anion Exchange Columntype HiScreen Capto Q w Column volume 4657 mi Pressure limit pre column 0 50 MPa 10 02 20 00 Pressure limit delta column 0 30 MPa 0 02 20 00 V Use flow restrictor 2 Attach an appropriate column holder to the rail on the instrument 3 Attach the column to the column holder 84 Getting Started with KTAavant and UNICORN 6 28 9440 69 AB 7 Prepare the system for a run 7 4 Connect a column Step Action 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 Instructions Result The Manual instructions dialog opens 6 In 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 Parameters for Column position 1 Pumps and pressures a Posion 1 Flow path _ a Injection valve Position 2 TJA Column position Flow Direction A Inlet A 1 Inlet B Down flow O Up flow pH valve d Sample inlet s Outlet valve Injection mark Monitors C Fraction collection 3 ty ar P Save resultas d 2
42. d after spillage If there is a risk that large volumes of spilled liquid may penetrate the casing of the AKTAavant instrument immediately switch off the instrument disconnect the power cord and contact an authorized service engineer 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 Superloop 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 maintenance e Hazardous biological agents during run When using hazardous biological agents run System CIP and Column CIP to flush the entire system tubing with bacteriostatic solution e g NAOH followed by a neutral buffer and finally distilled water before service and maintenance e RPC runs with 100 acetonitrile Always replace the green PEEK tubing between the used system pump and the 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 Getting Started with AKTAavant and UNICORN 6 28 9440 69 AB 17
43. e text by bold text e g Power switch Text entries that UNICORN generates or that the user must type are represented by a monotype typeface e g Program Files GE Healthcare UNICORN bin UNICORN Instrument Server exe config Getting Started with AKTAavant and UNICORN 6 28 9440 69 AB 9 1 About this guide 1 2 Regulatory information 1 2 Regulatory information Introduction This section describes the directives and standards that are fulfilled by the AKTAavant system CE conformity This product complies with the European directives listed in the table below by fulfilling the corresponding harmonized standards A copy of the Declaration of Conformity is available on request Directive Title 2006 42 EC Machinery Directive 2006 95 EC Low Voltage Directive 2004 108 EC ElectroMagnetic Compatibility EMC Directive 1999 5 EC Radio Equipment and Telecommunications Terminal Equipment Directive CE marking cE The CE marking and the corresponding declaration of conformity is valid for the instrument when it is e used as a stand alone unit or e connected to other CE marked GE Healthcare instruments or e connected to other products recommended or described in the user documentation and e used inthe same state as it was delivered from GE Healthcare except for alterations described in the user documentation 10 Getting Started with AKTAavant and UNICORN 6 28 9440 69 AB Interna
44. e the view of the different panes Illustration of the System Control user interface In the System Control module four panes show by default see illustration and table below 106 f Kee fie ka TER 25mm f ie moans NAN 5 Er Upa 5 mrar Ga east 5 pa 5 p 5 5 Crotoy x gay E wesesassesueessas o os Rum Log Piter on 2 20 mi System flow 1 00D biin Pre cotan menue iuad Procernel 2 28 mi System flow 1 000 ihin Pe cohen pense Comeletedt DA mi Bloch Slat Aat Ebdon Peeuedl Procerergl Completed 22 mi bane SeretoMan 2N mi Fiseteraten Volume 3 mi ides 01 n Newt tube Preusd Procenered ZN el Frecterwten Volume 3 mi tuber 01 led New Abe KCompietedt T pi Last tube ted Pane jauat Procesung 222 ni Lat hte fied Pana Completed 223 of End_Bock Gund Processing Comcieted 222 mt Woe Lren goderti Pinaed Procesung Conieted I mi Dae Saruhan 2 22 mi Geadert 100 00 8B 10 00 ICV rued ocesung I mi Geactere 100 00 R0 10 00 CV Congirted AZI mi Lant tb fiad The yatan all pae ahar the delay vahana um baen collected Fiysiend 02 s os Getting Started with KTAavant and UNICORN 6 28 9440 69 AB 8 Runa method 8 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
45. em 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 bll Phase Properties Text Instructions iT Sample Application Use the same flow rate as in Method Settings Flow rate 10 000 ml min 0 000 25 000 Iniect sample from loop Fill the loop using Manual load Loop type Capillary loop x ml O Inject sample directly onto column a ay Sample inlet Fill loop with ml Empty loop with 100 mi Sample volume ml C Use the same inlets as in Method Settings Ineta AT x InletB B1 oo z _ Fill the system with the selected buffer Getting Started with AKTAavant and UNICORN 6 28 9440 69 AB 8 Runamethod 8 2 Applying the sample 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 How to fill a capillary loop Follow the instruction below to fill the capillary loop with sample Step Action al Connect a suitable capillary loop to Injection valve ports LoopF fill and LoopE empty 2 Fill a syringe with sample 3 Connect the syringe to the Injection valve port Syr Getting Started with KTAavant and UNICORN 6 28 9440 69 AB 99 8 Runa method 8 2 Applying the sample Step Action 4 Load sample into the capillary loop To avoid sample loss due to siphoning
46. emove 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 Preparation 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 Library if desired Getting Started with AKTAavant and UNICORN 6 28 9440 69 AB 6 1 3 Predefined phases 6 Create a method 6 1 Guide to method creation 6 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 common parameters used in the subsequent phases Equilibration Equilibrates the column before purification or re equilibrates the column after purification Sample Application Applies sample to the column Defines the sample application technique the sample volume and the handl
47. ent 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 108 Getting Started with AKTAavant and UNICORN 6 28 9440 69 AB Customize Run Log 8 Runa method 8 4 Monitor the run 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 e Click the Customize icon to open the Customize dialog v4 e Under the Run Log tab select the items to display Customize System3 RunDataGroups Run Data Color Curves Curve Style and Color XAxis Y Axis Run Log
48. ers Follow the instruction below to rename files and folders in the Method Navigator or in the Result Navigator Step Action 1 Select the item that you want to rename Z Choose Edit Rename Result The current name is selected 5 Type a new name Getting Started with KTAavant and UNICORN 6 28 9440 69 AB 53 5 Files and folders in UNICORN 5 2 Handling files and folders Step Action 4 Press Enter Result The selected item is renamed Find files and folders Follow the instruction below to use the Find dialog to search for files or folders in the Method Navigator or in the Result Navigator Step Action 1 e Select the file or folder in the navigator pane from where you want to perform the search e Choose Edit Find Result The Find dialog opens Find what anion Find Nea Direction C Match whole word ony O Up Shift F3 C Match case Down F3 2 e Type a name or a partial name in the Find what field e Select none one or both of the the matching levels described below Match whole word only Matches only whole words and disregards partial matches Match case Matches the case of the Find what field entry e Select a Direction for the search Up The search proceeds upwards in the folder structure from the selected level Down The search proceeds downwards in the folder structure from the selected level 3 Click Find Next Result The first located match
49. et filters Clean the check valves Replace check valves Replace pump piston seals Replace pump pistons Replace pump rinsing system tubing Replace valve modules Getting Started with KTAavant and UNICORN 6 28 9440 69 AB 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 AKTAavant Drop Design and UNICORN are trademarks of GE Healthcare companies All third party trademarks are the property of their respective owners 2009 General Electric Company All rights reserved First published Sep 2009 All goods and services are sold subject to the terms and conditions of sale of the company within GE Healthcare which supplies them A copy of these terms and conditions is available on request Contact your local GE Healthcare representative for the most current information UN CORI Any use of this software is subject to GE Healthcare Standard Software End User License Agreement for Life Sciences Software Products GE Healt hcare Europe GmbH Munzinger Strasse 5 D 79111 Freiburg Germany GE Amersha GE Healt Healt hcare UK Limited m Place Little Chalfont Buckinghamshire HP7 9NA UK hcare Bio Sciences Corp 800 Centen
50. example port 2B 11 Click the End icon in the System Control toolbar to end the run Getting Started with AKTAavant and UNICORN 6 28 9440 69 AB 87 7 Prepare the system for a run 7 5 Calibrate the pH monitor 7 5 Calibrate the pH monitor Introduction If pH will be measured during the chromatographic run the pH monitor should be calibrated 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 88 Follow the instruction below to perform the calibration Step Action 1 Open the System Control module and select System Calibrate Result The Calibration dialog opens Calibration System3 Morte carate Tv Coren vale Calibration procedure Prepare for calibration Calibration for pH electrode buffer 1 Description pH calibrates the pH electrode with two point BH for buffer 1 calibration i e using two pH calibration solutions It 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 buffers E pH 4 or 7 as the
51. 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 ti i z 5 pH for buffer 1 Assymetty potential at pH 7 mV bed fo a J v 2 Set the pH monitor as the Monitor to calibrate by selecting pH from the list 5 Click Prepare for calibration 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 Getting Started with KTAavant and UNICORN 6 28 9440 69 AB 7 Prepare the system for a run 7 5 Calibrate the pH monitor Step Action 5 Fill a syringe with approximately 10 ml of the first pH standard buffer Connect the syringe to the Luer connector in pH valve port Cal and inject the buffer i l When the Current value is stable click rg Wash the 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 c
52. flush the entire system tubing with distilled water before service and maintenance WARNING Hazardous biological agents during run When using hazardous biological agents run System CIP and Column CIP to flush the entire system tubing with bacteriostatic solution e g NaOH followed by a neutral buffer and finally distilled water before service and maintenance Getting Started with AKTAavant and UNICORN 6 28 9440 69 AB 97 8 Runa method 8 2 Applying the sample 8 2 Applying the sample Introduction 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 applications techniques see User Manual Stage Description Load The capillary loop is filled with sample Inject The sample is injected onto the column Sample application through a capillary loop 98 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 syst
53. g un 05 ting 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 O Enter ID Select ID 28 9288 13 28928813 0000 00 1234 HiPrep 261 28 3286 13 12345678 0000 00 0005 HiPrep 2610 Desalting E Disable column logging 28 3288 13 12345678 0000 00 0006 HiPrep Desalting 2610 E 100 00 1234 HiPrep 2610 Desalting E Apply to all methods with the same column type Column Barcode 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 AKTAavant and UNICORN 6 28 9440 69 AB 105 8 Runa method 8 4 Monitor the run 8 4 Monitor the run Introduction You may follow the on going 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 customiz
54. gator may also show items that primarily are used in another UNICORN module The options are e Default Filter only items belonging to the active module show In Method Editor and System Control Methods Method Queues In Evaluation Results DoE Results e All all items show regardless of the active module e All except Archived only items that are not archived show The illustration below shows the filter settings available in the Method Navigator Method Navigator Open amp Methods Method Qu ly E Methods Method Queues Folder name All EH erug all except Archived Ell Method 1 Close the navigator pane 50 Click the small cross in the top right hand corner of the pane to close the navigator pane Getting Started with AKTAavant and UNICORN 6 28 9440 69 AB 5 Files and folders in UNICORN 5 2 Handling files and folders 5 2 Handling files and folders Introduction The handling of files and folders in UNICORN is similar to the general Windows functions This section describes how to create new folders and how to copy move delete and rename both files and folders It also describes the general search function that helps you find the files and folders that you are looking for Create a folder To arrange your files for better overview you can create sub folders in any folder that you have access to Follow the instruction below to create a new folder in the Method Navigator or in the Result
55. he selected system pump and repeat steps 3 4 82 Getting Started with KTAavant and UNICORN 6 28 9440 69 AB 7 Prepare the system for a run 7 4 Connect a column 7 4 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 AKTAavant WARNING Before connecting a column to the AKTAavant instrument read the instructions for use of the column To avoid exposing the column to excessive pressure make sure that the pressure limit is set to the specified maximum pressure of the column Methods automatically include a pressure alarm based on the tolerance of the chosen column type However when running manual runs you have to set the pressure limits yourself CAUTION Do not overtighten when connecting columns Overtightening might rupture the connectors or squeeze the tubing and thereby result in high backpressure Getting Started with AKTAavant and UNICORN 6 28 9440 69 AB 83 7 Prepare the system for a run 7 4 Connect a column Attach a column holder and connect a column to the instrument 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 appropriate tubing and connectors Step Action 1 Choose a column a
56. he 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 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 included 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 solution 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 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 AKTAavant and UNICORN 6 28 9440 69 AB 111
57. her procedures using solvent based buffers collect fractions through the outlet valve Getting Started with AKTAavant and UNICORN 6 28 9440 69 AB 71 7 Prepare the system for a run 7 2 Prepare the flow path 7 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 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 W1_ Column 4 PreC 0 00 MPa DeltaC 0 00 MPa UV 1 0 27 MAU 0 00 mS cm pH 0 00 Inlet B B 1 72 Getting Started with AKTAavant and UNICORN 6 28 9440 69 AB 7 Prepare the system for a run 7 2 Prepare the flow path Part Description 1 Sample inlet valve 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 Getting Started with KTAavant and UNICORN 6 28 9440 69 AB 73 7 Prepare the system for a run 7 2 Prepare the flow path Tubing and connectors
58. hod 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 40 Getting Started with AKTAavant and UNICORN 6 28 9440 69 AB 4 The instrument and the 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 gradient used in the method is displayed in a Gradient illustration 4 Cohn yee Hho Cato ateme 1 nf x Camme Tip EON cam wen Sam ie Appixaton Psm epeen 0 PRR Unk sion 2 Frenne ink dotaci 030 Mah 20009 3 a baan EZ Use fom seston Column Wash Flom Fate Une an v ed Cohan gestion Penton OPE tamea seecae See Wavelerghe 190 700 em Pom vate 100 nin M00 2 000 we Dm z n Corte te fw te peed overens Owe Couma Wash ows lee ee X Use maruah pepsed tutes B 2 Cable ph montong w a ww et ee atea C Um asma tafa mapaa Bafo Enable mt sereas sae Fece Brna a Bries O Sange riat 1 55 89 becomen n n Coe Deinet iy mpe hr suivonponm
59. hromatography e you have read and understood the Safety instructions chapter in the user documentation e the instrument and software are installed configured and calibrated according to AkTAavant and UNICORN 6 Installation Guide e a user account has been created according to UNICORN 6 Administration and Technical Manual 6 Getting Started with AKTAavant and UNICORN 6 28 9440 69 AB 1 About this guide 1 1 Get started 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 KTAavant and UNICORN 6 Installation Guide administration of UNICORN 6 Administration and Technical Manual databases instrument modulesand KTAavant and UNICORN 6 User Manual functions calibration KTAavant and UNICORN 6 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 AKTAavant and UNICORN 6 28 9440
60. iew 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 128 Getting Started with AKTAavant and UNICORN 6 28 9440 69 AB 9 Evaluate and print the results 9 3 Print the results Step Action 5 In the Customize Report dialog click the Print icon Result The Print dialog opens Properties Printer Microsoft Office Document Image Writer ts 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 report is printed Getting Started with AKTAavant and UNICORN 6 28 9440 69 AB 129 10 Maintenance 10 Maintenance Introduction This chapter provides schedules for preventive maintenance that should be performed by the user of AKTAavant Regular maintenance is essential for reliable function and results Refer to AKTAavant and UNICORN 6 User Manual for detailed instructions WARNING Always use appropriate personal protective equipment during operation and maintenance of AKTAavant system Contents This chapter contains the following sections Section See page 10 1 Maintenance program 131 130 Getting Sta
61. iles All users have a designated home folder where all methods and result files created by the user will automatically be located The files and folders are displayed in Navigator panes This chapter briefly describes the Navigator panes available in the different modules It also describes how to handle the files and folders This chapter contains the following sections Section See page 5 1 Navigator overview 48 5 2 Handling files and folders 51 Getting Started with AKTAavant and UNICORN 6 28 9440 69 AB 47 5 Files and folders in UNICORN 5 1 Navigator overview 5 1 Navigator overview Introduction This section describes the navigator panes that are available in all UNICORN modules with exception for the Administration module The navigator panes can be used to locate and open methods and results and to handle files and folders In the modules Method Editor and System Control the navigator panes are called Method Navigator and are used primarily to access methods In the Evaluation module the navigator pane is called Result Navigator In this navigator pane three different panes can be used to access the results Results Recent Runs and Find Results Show the navigator pane 48 To open and display the navigator pane in any of the UNICORN modules click the Open toolbar icon The navigator panes may either be displayed statically in the position where they open or the Auto Hide function can be selected by
62. information concerning the decommissioning of equipment materials in excess of the limits established by the Chinese 207 standard SJ T11363 2006 Requirements for Concentration This symbol indicates that the product contains hazardous waste and must be collected separately Please contact an authorized representative of the manufacturer for Limits for Certain Hazardous Substances in Electronic Information Product 22 Getting Started with AKTAavant and UNICORN 6 28 9440 69 AB 2 Safety instructions 2 3 Emergency procedures 2 3 Emergency procedures Introduction This section describes how to do an emergency shutdown of the AKTAavant system The section also describes the result 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 Getting Started with AKTAavant and UNICORN 6 28 9440 69 AB 23 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 AkTAavant instrument e The run is interrupted immediately e The data collected u
63. ing of flowthrough Column Wash Washes out unbound sample after sample application or removes strongly bound proteins after elution Performance Test Elution Elutes the sample from the column Defines parameters for the elution and fractionation settings Column Prepares the column before use by removing the storage Preparation solution 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 proteins By adding steps several cleaning solutions can be used sequentially System Prepares the system before a run by removing storage solution Preparation 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 Tests the efficiency of a packed column in terms of height equivalent to a theoretical plate HETP and the peak asymmetry factor AJ Getting Started with AKTAavant and UNICORN 6 28 9440 69 AB 67 6 Create a method 6 2 Print a method 6 2 Print a 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 yo
64. k integration 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 mA f e Under the Peak Table tab Select which data to display in the peak tables from the Select peak table columns list Click OK to display the settings and close the dialog Customize Chrom 1 Curve Style and Color Tf Edit Texts Layout Library Header CurveNames Y Anis Xsis Cuve Peak Table Select table to display Select peak table columns A Z Peak name Peaktable 4 7 Retention Peatere es endpoint retentions 7 rea Peaktable D P f total Peaktable E ercent of total area Peaktable F Percent of total peak area Peaktable G Height Peaktable H Width Peaktablet Width at half height Peaktable J Sigma Peaktable K Fraction tuba id Peaktable L fecuon SURA H Peaktable M Type of peak limits Peaktable N Peak endpoint heights Filter Peaks Fill settings Height less than 0 Fil peaks Width less than Pattern C Area less than C Width more than L T Peak must be one of 20 largest Do not show integration summary C Apply to all chromatograms Getting Started with AKTA
65. lick Eeue Result A pump wash of the first inlet in the Instruction execution list starts Getting Started with KTAavant and UNICORN 6 28 9440 69 AB 79 7 Prepare the system for a run 7 3 Prime buffer inlets and purge System pumps 80 Step Action 5 Connect a 25 30 ml syringe to one of the purge valves of the selected system pump Make sure that the syringe fits tightly into the purge connector 6 Open the purge valve by turning it counterclockwise about 3 4 of a turn Draw 5 10 ml of liquid slowly into the syringe 7 Close the purge valve by turning it clockwise Disconnect the syringe and discard its contents 8 Click the Continue button on the Instrument display to start a pump wash of the next inlet in the Instruction execution list 9 Repeat steps 4 7 until all inlets in the Instruction execution list have been primed filled with liquid Getting Started with AKTAavant and UNICORN 6 28 9440 69 AB 7 Prepare the system for a run 7 3 Prime buffer inlets and purge System pumps Purge the system pumps Follow the instruction below to purge System pump A and or System pump B Note that both pump heads of each system pump have to be purged Also note that both system pumps have to be purged after priming of buffer inlets TIP Prime all buffer inlets and purge System pump B before finally purging System pump A with start buffer The instrument is then ready for start of a method run Step Action 1
66. lose 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 AKTAavant and UNICORN 6 28 9440 69 AB 89 7 Prepare the system for a run 7 6 Prepare the Fraction collector 7 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 AKTAavant and UNICORN 6 User Manual WARNING Fraction collector Do not fractionate flammable liquids When running RPC methods or other procedures using solvent based buffers collect fractions through the outlet valve Prepare the Fraction collector Follow the instruction below to prepare the Fraction collector before a run Choose deep well plates and or tubes according to the fractionation settings in the method to be run and choose appropriate cassettes Step Action 1 If you are to use cassettes with the QuickRelease function open the cassettes 90 Getting Started with AKTAavant and UNICORN 6 28 9440 69 AB 7 Prepare the system for a run 7 6 Prepare the Fraction collector Step Action 2 Place the tubes and deep well
67. ment during operation and maintenance of AKTAavant system Hazardous substances When using hazardous chemical and biological agents take all suitable protective measures such as wearing protective glasses and gloves resistant to the substances used Follow local and or national regulations for safe operation and maintenance of the system Getting Started with AKTAavant and UNICORN 6 28 9440 69 AB 2 Safety instructions 2 1 Safety precautions e Spread of biological agents The operator has to take all necessary actions to avoid spreading hazardous biological agents in the vicinity of the instrument The facility should comply with the national code of practice for biosafety e High pressure The AKTAavant instrument operates under high pressure Wear protective glasses at all times Cautions CAUTION To avoid hazardous situations when working with the AKTAavant system take the following measures for personal protection 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 Cutinjuries The tubing cutter is very sharp and must be handled with care to avoid injuries Installing and moving the instrument Warnings WARNING To avoid personal injury when installing or moving the AKTAavant instrument follow the instructions below e Heavy object The AKTAavant instrument weighs about
68. n 6 1 1 Create and edit methods Step Action 8 Click the Save the method icon Result The Save As dialog opens Save As Methods Eh Folder name System Last modified Created by J UNICORN Database E Anita 01 14 2009 12 35 Default Anna 12 01 2008 14 12 lotta B Anne 12 01 2008 14 06 lotta B Bims 01 14 2009 12 42 Default B Bimo 01 14 2009 12 44 Default E Christer 12 01 2008 14 38 Default E Christine 12 02 2008 17 54 Default E Claes 11 21 2008 08 32 Default amp DefaultHome 09 25 2008 13 07 System SAsaDH 01 12 2009 15 56 Default ES Name UNTITLED System System3 o 9 In the Save As dialog e Select a target folder to enable the Save button e Type a Name for the method e Select a System from the list e Click the Save button Save Result The created method is saved in the selected folder 62 Getting Started with AKTAavant and UNICORN 6 28 9440 69 AB 6 Create a method 6 1 Guide to method creation 6 1 1 Create and edit methods Add phases to a method Follow the instruction below to add phases to a method Step Action 1 Select the desired phase in the Phase Library pane 2 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 Delete phases fr
69. nd UNICORN 6 28 9440 69 AB 119 9 Evaluate and print the results 9 2 Peak integration 9 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 integration 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 120 Getting Started with AKTAavant and UNICORN 6 28 9440 69 AB 9 Evaluate and print the results 9 2 Peak integration Step Action 3 In the Peak Integrate dialog e Select which curve to integrate 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 Peak Integrate Chromatogram Target peak table Chrom 1 001 U hrom 1 Manual Run 001 002 UV 2_0_Chrom 1 Manual Run 001 003 UY 3_0_Chrom 1 Manual Run 001 004 Cond_Chrom 1 Manual Run 001 005 Cond_Chrom 1 Manual Run 001 006 Conc B_Chrom 1 Manual Run 001 007 pH_Chrom 1 Manual Run 001 008 System flow_Chrom 1 Manual Run 001 009 System pressure_Chrom 1 Manual Run 001 010
70. nd UNICORN 6 28 9440 69 AB 101 8 Runa method 8 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 Disable column logging for this run Apply to all methods with the same column type Methods Remark Column Barcode Column type C g grun 1 ng 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 Result The first New Column dialog opens New Column exp ID came O O C C The Column has a uniTag has a fixed Code and exp Continue Cancel 102 Getting Started with KTAavant and UNICORN 6 28 9440 69 AB 8 Runa method 8 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 towa
71. nial Avenue P O Box 1327 Piscataway NJ 08855 1327 USA GE Healt hcare Bio Sciences KK Sanken Bldg 3 25 1 Hyakunincho Shinjuku ku Tokyo 169 0073 Japan 28 9440 69 AB 09 2009
72. om 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 Result 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 Getting Started with AKTAavant and UNICORN 6 28 9440 69 AB 63 6 Create a method 6 1 Guide to method creation 6 1 1 Create and edit methods Choose a maintenance method 64 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 Prepare tubing and instrument modules before a run Phase Method System Preparation Solution suitable buffers Prepare column before a run Column Preparation suitable buffers Clean tubing and modules System CI
73. on provides an overview of the AKTAavant instrument Technical details about the instrument and the individual modules are found in AKTAavant and UNICORN 6 User Manual Exterior design AkKTAavant 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 instrument 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 instrument s operational limits Parameter Limits Flow rate Up to 25 ml min Max operating pressure 20 MPa Wavelength 190 700 nm 32 Getting Started with AKTAavant and UNICORN 6 28 9440 69 AB 4 The instrument and the software 4 1 AKTAavant instrument overview Illustration of the main parts of the instrument The illustration below shows the location of the main parts of the instrument Part Description 1 Fraction collector 2 Buffer tray 3 Instrument display 4 Wet side 5 Foldable door 6 Pump cover 7 Holder rails 8 Swivel foot Lock Unlock knob 9 Swivel foot 10 Swing out toolbox 11 Power switch
74. oncsention Recos m 20 70 boconmendedi Coc Dsfned by tecipe har mubicorponent tasters Bastoni Vaeraties Getting Started with KTAavant and UNICORN 6 28 9440 69 AB Stat Protocol eo Und selection Metodat v x Flow Rate Uet en Morda etre Wavelerashe 980 700 rm wr 20 rm Owe m Dw E Latio sh nontong Eratie on soroa in Dira Eirin D Sanok riet Colam Loge Erte kozga O Ciian Patonin Tout oor 59 6 Create a method 6 1 Guide to method creation 6 1 1 Create and edit methods 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 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 60 Getting Started with AKTAav
75. ou 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 114 Getting Started with AKTAavant and UNICORN 6 28 9440 69 AB 9 Evaluate and print the results 9 Evaluate and print the results Introduction 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 Contents This chapter contains the following sections Section See page 9 1 View the results 116 9 2 Peak integration 120 9 3 Print the results 125 Getting Started with AKTAavant and UNICORN 6 28 9440 69 AB 115 9 Evaluate and print the results 9 1 View the results 9 1 View the results Introduction A result holds a complete record of the run including method system settings chromatogram 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 Step Action 1 Open the Evaluation module and click the Open Result Navigator icon Result The Result Navigator opens Result Navigator Results Recent Runs Find Results lv Open Gp Results DoE Results Folder name System Sij Eric amp Methods Results fa Run System3 fh Runz System3 2 Select the Results tab
76. own list 3 Prepare for calibration Press Result The pH valve switches to the calibration position Getting Started with AKTAavant and UNICORN 6 28 9440 69 AB 8 Runamethod 8 5 After run procedures Step Action 4 Prepare at least 10 ml storage solution by mixing equal volumes of a standard buffer pH 4 and a 1 M Potassium Nitrate KNO3 solution 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 6 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 log off UNICORN select File Log off Log off Eric Exit UNICORN Result All open UNICORN modules close and the Log On dialog opens Getting Started with AKTAavant and UNICORN 6 28 9440 69 AB 113 8 Runa method 8 5 After run procedures If you want to then exit UNICORN select 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 y
77. p to the time of the power failure is available in UNICORN 24 Getting Started with AKTAavant and UNICORN 6 28 9440 69 AB 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 connected 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 eliminate data loss after a power failure and allow time for a controlled shut down of the AKTAavant system Getting Started with AKTAavant and UNICORN 6 28 9440 69 AB 25 3 Start up 5 Start up Introduction Contents 26 This chapter describes how to start the system both the software and the instrument and also how to log off and exit UNICORN 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 This chapter contains the following sections Section See page 3 1 Start the system and log on to UNICORN 27 Getting Started with AKTAavant and UNICORN 6 28 9440
78. pose of the guide and provides regulatory information relevant for AKTAavant system Contents This chapter contains the following sections Section See page 1 1 Get started 6 1 2 Regulatory information 10 Getting Started with AKTAavant and UNICORN 6 28 9440 69 AB 5 1 About this guide 1 1 Get started 1 1 Get started Introduction Read this section for an understanding of the purpose and conventions of this guide as well as the requirements that you must fulfill before using the AKTAavant system 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 AKTAavant 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 AKTAavant 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 e you understand the concepts of liquid c
79. pped before opening the drawer Getting Started with AKTAavant and UNICORN 6 28 9440 69 AB 93 7 Prepare the system for a run 7 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 HH ESSE eo eoe EEETAERNEEER 0 8 0 9 o 23 PY INOS HO1937109 NOILOVHS ee oe eeeo ee ee O Gus 000o 000o 94 Getting Started with KTAavant and UNICORN 6 28 9440 69 AB 8 Runa method 8 Runa method Introduction This chapter describes how to start up and run a method and also how to handle the system after the run Contents This chapter contains the following sections Section See page 8 1 Before you start 96 8 2 Applying the sample 98 8 3 Start a method run 101 8 4 Monitor the run 106 8 5 After run procedures 111 Getting Started with AKTAavant and UNICORN 6 28 9440 69 AB 95 8 Runa method 8 1 Before you start 8 1 Before you start Introduction Checklist Before starting a run it is necessary to read and understand the information in this section and to perform the checks listed below Make sure that the system is correctly prepared Check that the system is prepared according to chapter 6 a suitable column has been selected for the application consider target protein and pressure range the
80. rds 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 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 t Alias optional i o Technique Desalting Column type HiPrep 26 10 Desalting C Use medium batch ID Getting Started with KTAavant and UNICORN 6 28 9440 69 AB 103 8 Runa method 8 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 click OK TIP Alias can be used for easy identification of a column Result The entered information is saved and the dialog closes 104 Getting Started with KTAavant and UNICORN 6 28 9440 69 AB 8 Runa method 8 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 lot exp ID Enter ID O Select ID 28 9298 13 12345676 0000 00 0005 HiPrep 261 Disable column logging for this run Apply to all methods with the same column type Methods Column type Column Handlin
81. rted with AKTAavant and UNICORN 6 28 9440 69 AB 10 1 Maintenance program Introduction 10 Maintenance 10 1 Maintenance program This section describes the preventive maintenance to be performed on AKTAavant The maintenance is divided into Daily maintenance Weekly maintenance Monthly maintenance Bi annual maintenance Maintenance when required Periodic maintenance program The following periodic maintenance should be performed by the user of AKTAavant Interval Maintenance action Daily Calibrate the pH monitor Weekly Calibrate pressure monitors Weekly Change pump rinsing solution Monthly Check the flow restrictor Bi annual Clean the UV flow cell Bi annual Replace pH electrode Getting Started with KTAavant and UNICORN 6 28 9440 69 AB 131 10 Maintenance 10 1 Maintenance program Maintenance when required The following maintenance should be performed by the user of AKTAavant when required Maintenance action 132 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 Replace online filter Replace o ring in mixer Replace UV flow cell Replace flow restrictor Replace inl
82. sssssssssssssssssssssssssssssssssssssssssasstnnsnsnnssssntnsnnnntnnssnsnnnensesseseseeeeee 43 4 3 3 Evaluation module wieeeccecccssescssssessssssesssssssssssssesssssesecssssssssssssecsssssssssssueessssssessssuesssauesssssnesssssseeessane 45 5 Filesand folders in UNICORN vscisscossessiscisveesscssecossezsvscsesasesctaveosedesbssssveevsassesessasses 47 5 1 Navigator OVeFV EW weccececssssssccsssssssesscssssussccscsssssessssssssssecescessuusecsssssssesscssssssusesesssssusesesssssueesecesssasecs 48 52 HCA AIAG FIIES GA FOVA STS siniseen nA anna 51 GB Cregted Method essorer rees sE r r E ENE E EES 55 6 1 Guide to method creation wa seeeeceseeeescssssesssssessssssesessssssssssssesssssuesssssuessssseesssssucsssssusecssseesessneesesssuesessee 56 6 1 1 Create and edit methodS ss sssisseisssrssesrsesrsserssrressrrssrrssresstrrsnnreserrsnnrrsntrrsnnrsnerrsntnrntrrnnnrne 57 61 2 Predefingd Methods issiseiniieniarnoiiinaniiiie i ea ATE 65 61 3 Predefined phases irinosisuiiiirinimenuiiniii ig E 67 62 Printa methodi ar EE E 68 7 Prepare the system fof aA FUN is dicsnidea ki decccchsdecrericcrshstiaeatiaaedcennectehtekitnecet 70 7 1 Before you prepare the SyStem u c eccsssssssssesssscccsssssssssssssessssssecsssssssssessssessesesssssssuusessssscessssesssees 71 Te Prepare the Now pathi sins ssccsscvsetaseecictccsssseesssasnavssuchenteGhevuvsssslastsosebescuvstsosiutbebbeasbonssnvoivissiect etectoesties 72 7 3 Prime buffer inlets and purge System pumps 78
83. struction pane press the F1 key select Manual Execute Manual Instructions expand a heading and select the instruction of interest click the Help icon in the dialog Getting Started with AKTAavant and UNICORN 6 28 9440 69 AB 4 The instrument and the software 4 3 UNICORN software modules 4 3 UNICORN software modules Introduction Contents 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 This section contains the following subsections Section See page 4 3 1 Method Editor module 40 4 3 2 System Control module 43 4 3 3 Evaluation module 45 Getting Started with AKTAavant and UNICORN 6 28 9440 69 AB 39 4 The instrument and the 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 met
84. sult The Manual instructions dialog opens 78 Getting Started with AKTAavant and UNICORN 6 28 9440 69 AB 7 Prepare the system for a run 7 3 Prime buffer inlets and purge System pumps Step Action 4 TIP If desired the flow rate and volume of the pump wash can be edited before the pump wash is started In the Manual instructions dialog select Pumps and pressures Wash settings and edit the settings for System flow rate and System pump wash volume Click Execute In the Manual instructions dialog e Select Pumps and pressures Pump wash and select the inlet to be primed from the Inlet A or Inlet B drop down list Manual instructions System3 Instructions s ae 7 d arameters for Pump was Pumps and pressures A Inlet A 0 System flow 1 Sample flow At uE V 4 Gradient Inlet B 4 Pump wash f System wash Off g i Wash settings BufferPro Q inlets BufferPro pH r L Pressure control parameters Off vja Fow path Sample inlet G Monitors id Fraction collection Fe Off oe ia L Save result as Browse vV Auto update parameter fields will be updated during method run d e If several inlets are to be used click Insert to insert a selected inlet in the Instruction execution list Insert all inlets to be used in the Instruction execution list instruction execution list Pump wash A1 Off Off Off IP ff Off Off ash A3 Off Off Off Execute Close c
85. t Protocol System Calibration Evaluation Log Questions 0 00 min Method Run 2009 01 23 09 55 33 01 00 Method Dokumentation Result Moa Dokumentation Dokumentation 001 User Mia 0 00 min Batch ID sdfgfihertniztatnz 0 00 min Base Volume 20 106 ml GSTPrep FF 16 10 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 Base SameAsMain 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 Alarm delta column pressure Enabled 3 50 MPa 0 00 MPa Completed 0 01 min Wavelength 280 nm Off nm Off rm Issued Processing 0 02 min Wavelength 280 rm Off nm Off nm Completed 0 02 min Noise reduction UV 5 0 sec Issued Processing NN min Nanise rediiction HIV 5M feer Camnleted lt Adjust retention zero to injection number Included entry types Included feedback types Base Set Mark 7 Issued Time Block Start Processing O Volume Method Failed Manual Ignored v v Z Alarms Wamings and Errors X Completed v v v System Cancelled a Getting Started with AKTAavant a
86. t b r Column Leghook s Dawen i Varabies Getting Started with KTAavant and UNICORN 6 28 9440 69 AB 41 4 The instrument and the software 4 3 UNICORN software modules 4 3 1 Method Editor module Method Editor toolbar icons The table below shows the Method Editor toolbar icons that are referred to in this Getting Started guide Icon Function New Method Opens the New Method dialog where methods can be created Icon 5 Function Open Method Navigator Opens the Method Navigator where available methods are listed T Save Saves the active method Print Opens the Print dialog from where a method can be printed D Copy Copies the selected method or folder Paste Pastes a copied method or folder to a new location Ls 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 42 Getting Started with AKTAavant and UNICORN 6 28 9440 69 AB 4 The instrument and the 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 da
87. ta 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 2 pree ee ae i gt ine 250 t n 2m 0 1 O00 ete O23 MPs ars Via y Cond 55 snl 1 ears 5 ae 0157 wl EM sem 1 Fone la m hau oSUHEEESEBESE Pum Log Fater en 2 20 of System fw 1 000 inter Pre cohen permen Deued Placecure 2 21 mi Fisetoraten Vama 3 ni tuber 11 in Newt tde femal Pocmsrg N mi Fiacteraten Volume 3 mi biter 01 Intl New Ade Compietedt X mi Last tube tiled Pane Pienued Procesung P mi Lact kibe flod Pana Compared 223 ai End Boc Juua Pocensingt Completed 222 mt Block Lorem gader Pissed Pocesing Longines 222 mi Dave Semen 2 22 mi Grader 100 00 38 10 00 KV prud Pocesung 2 22 mi Graders 100 09 R0 10 00 CV Fongirted Getting Started with AKTAavant and UNICORN 6 28 9440 69 AB 43 4 The instrument and the 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 guide Icon Function Open Method Navigator Opens the Method Navigator where available methods are listed Icon Function Run Starts a method run Hold Suspends the method run while current flow rate and valve positions are sustained tal
88. tart buffer Anion Exchange Chromatography AIEX After equilibration and sample application negatively charged proteins are adsorbed to the column ligand After a wash to remove 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 Cation Exchange After equilibration and sample application positively charged Performance Test Chromatography proteins are adsorbed to the column ligand After a wash to CIEX remove 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 Chromatofocusing After equilibration and sample application elution is performed CF using a pH gradient The proteins separate and elute according to their isoelectric points Finally the column is re equilibrated 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 Suggestions for cleaning steps are available for a number of column types Column After equilibration of the column sample is injected via a capillary loop and eluted isocratically Anon adsorbing sample like acetone or salt should be used After the run calculate column performance in the Evalu
89. tional standards This product fulfills the requirements of the following standards Standard EN 61010 1 IEC CAN CSA C22 2 No 61010 1 61010 1 UL 61010 1 Description Safety requirements for electrical equipment for measurement control and laboratory use 1 About this guide 1 2 Regulatory information Notes EN 61326 1 EMC emissions and immunity requirements for electrical equipment for measurement control and laboratory use Harmonized with 2004 108 EC CISPR 11 Group 1 Class B 12100 2 EN ISO 12100 1 Safety of machinery Basic concepts general principles for design Harmonized with 2006 42 EC 14121 2 EN ISO 14121 1 Safety of machinery Principles of risk assessment Harmonized with 2006 42 EC 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 the KTAavant system 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 Getting Started with KTAavant and UNICORN 6 28 9440 69 AB 2 Safety instructions 2 Safety instructions Introduction Contents 12 This chapter contains instructions of ho
90. trument module is removed or installed or a cable is connected or disconnected 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 connectors Do not disconnect or move the UniNet 9 bus cable System operation Warnings WARNING To avoid personal injury when operating the AKTAavant system follow the instructions below 16 Rotating the instrument Make sure that there is always at least 20 cm of free space around the AKTAavant instrument to allow rotation on the swivel foot When rotating the system take care not to stretch or squeeze tubing or cables A disconnected cable may cause power interruption or network interruption Stretched tubes may cause bottles to fall resulting in liquid spillage and shattered glass Squeezed tubing may cause increase in pressure or block liquid flow To avoid the risk of turning over bottles always place bottles on the buffer tray and close the doors before rotating the instrument Getting Started with AKTAavant and UNICORN 6 28 9440 69 AB 2 Safety instructions 2 1 Safety precautions e Fasten sample bottles Always fasten bottles and cassettes to the rails at the sample tray Use appropriate holders for bottles Shattered glass from falling bottles may cause injury Spilled liquid may cause fire hazard and personal injury e Electrical shock hazar
91. ults 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 report 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 a suitable zoom factor has been selected Getting Started with AKTAavant and UNICORN 6 28 9440 69 AB 125 9 Evaluate and print the results 9 3 Print the results Print a chromatogram and a peak table Follow the instruction below to print a chromatogram and if peak integration has been performed the related peak table Step Action 1 Open the Evaluation module and open a result 2 Click the Print icon Result The Print Chromatograms dialog opens Print Chromatograms Printer Microsoft Office Document Image Writer K Print Format EO amp amp bY mm 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 Preview Result A preview of the chromatogram displays in the Customize Report
92. ur computer How to print a method Follow the instruction below to print a method 68 Step Action 1 Open the Method Editor module and click the Open Method Navigator icon in the toolbar Result The Method Navigator pane opens Method Navigator Open amp Methods Method Q Folder name System a Bec E Methods Method1 System3 Method2 Systema E Results 2 Select the method to print and click the Open a Method icon in the navigator toolbar Open Result The selected method opens Getting Started with AKTAavant and UNICORN 6 28 9440 69 AB 6 Create a method 6 2 Print a method Step Action 3 Click the Print icon Result The Print dialog opens Print Method 1 Printer Adobe PDF w Properties Print items Phase range T Text instructions All phases M Variable list O Phase Method Settings 4 In the Print dialog e select Printer e click OK Result The method is printed Getting Started with AKTAavant and UNICORN 6 28 9440 69 AB 69 7 Prepare the system for a run 7 Prepare the system for a run Introduction Follow this chapter to perform the necessary preparations of the system before starting a run WARNING Always use appropriate personal protective equipment during operation and maintenance of AKTAavant system Contents This chapter contains the following sections Section See page 7 1 Before you prep
93. w to handle the AKTAavant instrument in a safe way This chapter contains the following sections Section See page 2 1 Safety precautions 13 2 2 Labels 20 2 3 Emergency procedures 23 Getting Started with AKTAavant and UNICORN 6 28 9440 69 AB 2 Safety instructions 2 1 Safety precautions 2 1 Safety precautions Introduction The AKTAavant 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 General precautions Personal protection Installing and moving the instrument System operation Maintenance General precautions WARNING Always follow these General precautions to avoid injury when using the AKTAavant instrument Do not operate the AKTAavant instrument in any other way than described in the AKTAavant and UNICORN manuals Operation and user maintenance of the AKTAavant instrument should be performed by properly trained personnel only Before connecting a column to the AKTAavant instrument read the instructions for use of the column To avoid exposing the column to excessive pressure make sure that the pressure limit is set to the specified maximum pressure of the column

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