GE AKTAavant Getting Started
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GE AKTAavant Getting Started

GE AKTAavant Getting Started

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Getting Started with
ÄKTAavant and UNIC ORN 6
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Summary of Contents for GE AKTAavant

  • Page 1 GE Healthcare Getting Started with ÄKTAavant and UNIC ORN 6 ™ ™...

  • Page 3: Table Of Contents

    Table of Contents Table of Contents About this guide ....................Get started ............................... Regulatory information ........................Safety instructions ....................Safety precautions ..........................Labels ................................. Emergency procedures ........................Start up ......................... Start the system and log on to UNICORN .................. The instrument and the software ..............ÄKTAavant instrument overview ....................
  • Page 4 Table of Contents Monitor the run ............................After run procedures ........................... Evaluate and print the results ................View the results ............................. Peak integration ............................ Print the results ............................10 Maintenance ......................10.1 Maintenance program ........................Getting Started with ÄKTAavant and UNICORN 6 28-9440-69 AB...

  • Page 5: About This Guide

    1 About this guide 1 About this guide Introduction This chapter describes the purpose of the guide and provides regulatory information relevant for ÄKTAavant system. Contents This chapter contains the following sections: Section See page 1.1 Get started 1.2 Regulatory information Getting Started with ÄKTAavant and UNICORN 6 28-9440-69 AB...

  • Page 6: Get Started

    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 ÄKTAavant 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 ÄKTAavant instrument.
  • Page 7 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..
  • Page 8 1 About this guide 1.1 Get started Intended use ÄKTAavant 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 ÄKTAavant system is intended for research use only, and shall not be used in any clinical procedures, or for diagnostic purposes.
  • Page 9 Hardware items are identified in the text by bold text (e.g., Power switch). Text entries that UNICORN generates or that the user must type are represented by a monotype typeface (e.g., \Program Files\GE Healthcare\UNICORN\bin\UNICORN Instrument Server.exe.config). Getting Started with ÄKTAavant and UNICORN 6 28-9440-69 AB...

  • Page 10: Regulatory Information

    • used in the same state as it was delivered from GE Healthcare, except for alterations described in the user documentation. Getting Started with ÄKTAavant and UNICORN 6 28-9440-69 AB...
  • Page 11 1 About this guide 1.2 Regulatory information International standards This product fulfills the requirements of the following standards: Standard Description Notes EN 61010-1, IEC Safety requirements for electrical 61010-1, UL 61010-1, equipment for measurement, CAN/CSA-C22.2 control, and laboratory use No.61010-1 EN 61326-1 EMC emissions and immunity Harmonized with...

  • Page 12: Safety Instructions

    2 Safety instructions 2 Safety instructions Introduction This chapter contains instructions of how to handle the ÄKTAavant instrument in a safe way. Contents This chapter contains the following sections: Section See page 2.1 Safety precautions 2.2 Labels 2.3 Emergency procedures Getting Started with ÄKTAavant and UNICORN 6 28-9440-69 AB...

  • Page 13: Safety Precautions

    • Do not use any accessories not supplied or recommended by GE Healthcare. • Do not use the ÄKTAavant instrument if it is not working properly, or if it has suffered...
  • Page 14 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 ÄKTAavant instrument, follow these precautions to avoid any risk of fire or explosion. •...
  • Page 15 • Protective ground. The ÄKTAavant instrument must always be connected to a grounded power outlet. • Power cord. Only use power cords delivered or approved by GE Healthcare. Getting Started with ÄKTAavant and UNICORN 6 28-9440-69 AB...
  • Page 16 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. •...
  • Page 17 2 Safety instructions 2.1 Safety precautions • 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. •...
  • Page 18 2 Safety instructions 2.1 Safety precautions Cautions CAUTION To avoid hazardous situations when operating the ÄKTAavant 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.
  • Page 19 ÄKTAavant instrument, follow the instructions below. • Electrical shock hazard. All repairs should be done by service personnel authorized by GE Healthcare. Do not open any covers or replace parts unless specifically stated in the user documentation. • Disconnect power. Always switch off power to the instrument before replacing any component on the instrument, unless stated otherwise in the user documentation.

  • Page 20: Labels

    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 ÄKTAavant instrument. For information about marking of the computer equipment, refer to the manufacturer’s instructions. Labels on the ÄKTAavant instrument The illustrations below show the labels that are attached to the ÄKTAavant instrument.
  • Page 21 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.
  • Page 22 2 Safety instructions 2.2 Labels Labels concerning hazardous substances The following symbols on the labels concern hazardous substances: Label Meaning This symbol indicates that electrical and electronic equipment must not be disposed of as unsorted municipal waste and must be collected separately. Please contact an authorized representative of the manufacturer for information concerning the decommissioning of equipment.

  • Page 23: Emergency Procedures

    2 Safety instructions 2.3 Emergency procedures 2.3 Emergency procedures Introduction This section describes how to do an emergency shutdown of the ÄKTAavant 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...
  • Page 24 2 Safety instructions 2.3 Emergency procedures Step Action 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...
  • Page 25 2 Safety instructions 2.3 Emergency procedures Power failure to... will result in... Computer The UNICORN computer shuts down • The instrument display shows status Not • connected The run is interrupted immediately • Data generated up to 10 seconds before the •...

  • Page 26: Start Up

    3 Start up 3 Start up Introduction 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.

  • Page 27: Start The System And Log On To Unicorn

    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.
  • Page 28 3 Start up 3.1 Start the system and log on to UNICORN Step Action In the Log On dialog: Select your User Name from the drop-down list. • Enter your Password. • Click the Options button. • Result: Checkboxes for each of the UNICORN modules are displayed in the Log On dialog.
  • Page 29 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. Step Action Switch on the instrument by pressing the Power switch to the I position. Open the the System Control module in UNICORN and click the Connect to Systems icon.
  • Page 30 3 Start up 3.1 Start the system and log on to UNICORN Step Action In the Connect to Systems dialog: Select the instrument. • Select Control mode. • Click OK. • Result: The selected instrument can now be controlled by the software. Getting Started with ÄKTAavant and UNICORN 6 28-9440-69 AB...

  • Page 31: The Instrument And The Software

    4 The instrument and the software 4 The instrument and the software Introduction This chapter gives an overview of the ÄKTAavant system: instrument, software and accessories. Illustration of the system The illustration below shows the ÄKTAavant instrument with UNICORN software installed on a computer.

  • Page 32: Äktaavant Instrument Overview

    4 The instrument and the software 4.1 ÄKTAavant instrument overview 4.1 ÄKTAavant instrument overview Introduction This section provides an overview of the ÄKTAavant instrument. Technical details about the instrument and the individual modules are found in ÄKTAavant and UNICORN 6 User Manual.
  • Page 33 4 The instrument and the software 4.1 ÄKTAavant 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 Fraction collector Buffer tray Instrument display Wet side Foldable door Pump cover Holder rails...
  • Page 34 4 The instrument and the software 4.1 ÄKTAavant instrument overview Illustration of the wet side modules of the instrument The illustration below shows the modules of the wet side of the instrument. Part Description Injection valve Column valve Flow restrictor pH valve UV monitor Conductivity monitor...
  • Page 35 4 The instrument and the software 4.1 ÄKTAavant instrument overview Part Description System pump A Mixer Pressure monitor of Sample pump Sample pump Pump piston rinsing system tubes Sample inlet valve Outlet valve Holder rails Getting Started with ÄKTAavant and UNICORN 6 28-9440-69 AB...

  • Page 36: Unicorn Software Overview

    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 and Evaluation.
  • Page 37 4 The instrument and the software 4.2 UNICORN software overview 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...
  • Page 38 4 The instrument and the software 4.2 UNICORN software overview If you want to... then... 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 •...

  • Page 39: Unicorn Software Modules

    4 The instrument and the software 4.3 UNICORN software modules 4.3 UNICORN software modules Introduction 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.

  • Page 40: Method Editor Module

    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.
  • Page 41 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).
  • Page 42 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 Icon Function New Method: Opens the Open Method Navigator: New Method dialog where...

  • Page 43: System Control Module

    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 data as curves during the entire method run.
  • Page 44 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 Icon Function Open Method Navigator: Run: Starts a method run.

  • Page 45: Evaluation Module

    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.
  • Page 46 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 Icon Function Open Result Navigator: Save: Saves the changes Opens the Result Navigator made to the current result.

  • Page 47: Files And Folders In Unicorn

    5 Files and folders in UNICORN 5 Files and folders in UNICORN Introduction In the UNICORN user interface, method and result data are represented by files. All users have a designated home folder where all methods and result files created by the user will automatically be located.

  • Page 48: Navigator Overview

    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.
  • Page 49 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: Opens the selected file. New folder: Creates a new folder in the folder that is currently selected. Refresh: Updates all items in the navigator pane to the current status.
  • Page 50 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 navigator may also show items that primarily are used in another UNICORN module. The options are: •...

  • Page 51: Handling Files And Folders

    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.
  • Page 52 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.
  • Page 53 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: 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.
  • Page 54 5 Files and folders in UNICORN 5.2 Handling files and folders Step Action 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.

  • Page 55: Create A Method

    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...

  • Page 56: Guide To Method Creation

    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.

  • Page 57: Create And Edit Methods

    6 Create a method 6.1 Guide to method creation 6.1.1 Create and edit methods 6.1.1 Create and edit methods Introduction Method creation in UNICORN 6 is performed in the Method Editor module. Predefined methods are available for a number of different chromatography techniques, for example affinity chromatography and gel filtration.
  • Page 58 6 Create a method 6.1 Guide to method creation 6.1.1 Create and edit methods Part Description Phase Library: Displays all available phases. Method Outline: Presents an overview of the phases included in the active method. Phase Properties: Presents detailed settings for the selected phase. Gradient: Illustrates the gradient used in the active method.
  • Page 59 6 Create a method 6.1 Guide to method creation 6.1.1 Create and edit methods Step Action 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.
  • Page 60 6 Create a method 6.1 Guide to method creation 6.1.1 Create and edit methods Step Action In the Phase Properties pane of the Method Settings phase, click the Result Name & Location button to specify the name and location of the results from the method runs.
  • Page 61 6 Create a method 6.1 Guide to method creation 6.1.1 Create and edit methods Step Action In the Phase Properties pane of the Method Settings phase, click the Start Protocol button if you wish to include a start protocol. Result: The Start Protocol dialog opens. In the Start Protocol dialog: Select items to display at method start.
  • Page 62 6 Create a method 6.1 Guide to method creation 6.1.1 Create and edit methods Step Action Click the Save the method icon. Result: The Save As dialog opens. In the Save As dialog: Select a target folder to enable the Save button. •...
  • Page 63 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 Select the desired phase in the Phase Library pane. Drag-and-drop the phase to requested position in the Method Outline pane.
  • Page 64 6 Create a method 6.1 Guide to method creation 6.1.1 Create and edit methods Choose a maintenance method A number of predefined methods for preparation and cleaning are available. Use these maintenance methods to prepare and clean the system and columns, and to fill the system and columns with storage solution.

  • Page 65: Predefined Methods

    6 Create a method 6.1 Guide to method creation 6.1.2 Predefined methods 6.1.2 Predefined methods The predefined methods that are available are described in the table below. Method Description Affinity After equilibration and sample application, the protein of chromatography interest is adsorbed to the column ligand. After a wash to (AC) remove unbound sample, elution is performed either by using a buffer containing a competitor to displace the protein of...
  • Page 66 6 Create a method 6.1 Guide to method creation 6.1.2 Predefined methods Method Description Desalting (DS) After equilibration and sample application, the proteins are eluted isocratically. This technique is commonly used for buffer exchange. Gel filtration (GF) After equilibration and sample application, proteins separate and elute according to their size (largest first).

  • Page 67: Predefined Phases

    6 Create a method 6.1 Guide to method creation 6.1.3 Predefined phases 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.

  • Page 68: Print A Method

    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 your computer. How to print a method Follow the instruction below to print a method.
  • Page 69 6 Create a method 6.2 Print a method Step Action Click the Print icon. Result: The Print dialog opens. In the Print dialog: select Printer • click OK • Result: The method is printed. Getting Started with ÄKTAavant and UNICORN 6 28-9440-69 AB...

  • Page 70: Prepare The System For A Run

    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 ÄKTAavant system. Contents This chapter contains the following sections: Section...

  • Page 71: Before You Prepare The System

    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.

  • Page 72: Prepare The Flow Path

    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.
  • Page 73 7 Prepare the system for a run 7.2 Prepare the flow path Part Description Sample inlet valve Sample pump Inlet valve A System pump A Quaternary valve Inlet valve B System pump B Injection valve Capillary loop or Superloop Column valve Column UV monitor Conductivity monitor...
  • Page 74 7 Prepare the system for a run 7.2 Prepare the flow path Tubing and connectors The table below shows what types of tubing and connectors to use for different purposes. Scope of use Tubing Connector Photo Inlet tubing Teflon™, o.d. Tubing connector, 1/8"...
  • Page 75 7 Prepare the system for a run 7.2 Prepare the flow path Valves and ports Illustrations Inlet valve B Inlet ports: B1-B7 • Sample inlet valve Inlet ports: S1-S7, Buff • Getting Started with ÄKTAavant and UNICORN 6 28-9440-69 AB...
  • Page 76 7 Prepare the system for a run 7.2 Prepare the flow path Waste ports The table below shows the waste ports of Injection valve, pH valve, and Outlet valve. Valves and ports Illustrations Injection valve Waste ports: W1, W2 • pH valve Waste port: •...
  • Page 77 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 Connect inlet and outlet tubing to the ports that are to be used. Immerse all inlet tubing that is to be used during the method run in the correct buffers.

  • Page 78: Prime Buffer Inlets And Purge System Pumps

    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).
  • Page 79 7 Prepare the system for a run 7.3 Prime buffer inlets and purge System pumps Step Action If desired, the flow rate and volume of the pump wash TIP: 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.
  • Page 80 7 Prepare the system for a run 7.3 Prime buffer inlets and purge System pumps Step Action 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. Open the purge valve by turning it counterclockwise about 3/4 of a turn.
  • Page 81 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.
  • Page 82 7 Prepare the system for a run 7.3 Prime buffer inlets and purge System pumps Step Action 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. Open the purge valve by turning it counterclockwise about 3/4 of a turn.

  • Page 83: Connect A Column

    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 ÄKTAavant.
  • Page 84 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.
  • Page 85 7 Prepare the system for a run 7.4 Connect a column Step Action 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. Open the System Control module and select Manual:Execute Manual Instructions.
  • Page 86 7 Prepare the system for a run 7.4 Connect a column Step Action In the Manual instructions dialog: Select Pumps and pressure:System flow. • Enter a low Flow rate (e.g., 0.2 ml/min). • • Click Result: A system flow of 0.2 ml/min starts. 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.
  • Page 87 7 Prepare the system for a run 7.4 Connect a column Step Action Connect a tube to the bottom of the column. 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 example port 2B.

  • Page 88: Calibrate The Ph Monitor

    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.
  • Page 89 7 Prepare the system for a run 7.5 Calibrate the pH monitor Step Action 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.

  • Page 90: Prepare The Fraction Collector

    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 ÄKTAavant and UNICORN 6 User Manual.
  • Page 91 7 Prepare the system for a run 7.6 Prepare the Fraction collector Step Action Place the tubes and deep well plates in the cassettes. Make sure that the deep well plates are rotated so that the well marked A1 is positioned above the A1 marking on the cassette.
  • Page 92 Action 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.
  • Page 93 Action 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. Close the Frac drawer. Make sure that it snaps into closed position.
  • Page 94 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. Getting Started with ÄKTAavant and UNICORN 6 28-9440-69 AB...

  • Page 95: Run A Method

    8 Run a method 8 Run a 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 8.2 Applying the sample 8.3 Start a method run...

  • Page 96: Before You Start

    8 Run a method 8.1 Before you start 8.1 Before you start Introduction Before starting a run, it is necessary to read and understand the information in this section and to perform the checks listed below. Checklist Make sure that the system is correctly prepared. Check that: •...
  • Page 97 8 Run a method 8.1 Before you start If you want to... then... resume, for example, a held or paused click the Continue icon. method run. An ended method Note: cannot be continued. permanently end the run click the End icon. Note: When ending a method run in advance, it is possible to save the partial result.

  • Page 98: Applying The Sample

    8 Run a 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.
  • Page 99 8 Run a method 8.2 Applying the sample Empty the capillary loop with a buffer volume that exceeds the volume TIP: 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 Connect a suitable capillary loop to Injection valve ports LoopF (fill) and...
  • Page 100 8 Run a method 8.2 Applying the sample Step Action Load sample into the capillary loop. To avoid sample loss due to siphoning, leave the syringe in the port until the sample has been injected onto the column during the run. It is recommended to overload the loop to make sure that TIP: the loop is completely filled.

  • Page 101: Start A Method Run

    8 Run a 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.
  • Page 102 8 Run a method 8.3 Start a method run Step Action Click Finish on the last page of the Start Protocol. Result: 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.
  • Page 103 8 Run a method 8.3 Start a method run Step Action Register the column using the 2D Barcode scanner as follows: Make sure that the pointer is placed in the first position of the Code field. • Point the 2D Barcode scanner towards the data matrix tag on the column. •...
  • Page 104 8 Run a method 8.3 Start a method run Step Action In the expanded New Column dialog: enter Alias (optional) • select Technique and Column type from the drop-down lists. • click OK. • Alias can be used for easy identification of a column. TIP: Result: The entered information is saved and the dialog closes.
  • Page 105 8 Run a method 8.3 Start a method run Step Action In the Select Columns dialog: Select Enter ID. • Use the 2D Barcode scanner (see step 7) to enter the column ID. • Alternatively, choose Select ID and select the column individual to be •...

  • Page 106: Monitor The Run

    8 Run a 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.
  • Page 107 8 Run a method 8.4 Monitor the run Part Description Run Data: Presents current run data values. Chromatogram: Illustrates data as curves. Run Log: Presents all registered actions. Flow Scheme: Illustrates the current flow path. Customize Run Data The Run Data pane shows real-time data from available monitors during the run.
  • Page 108 8 Run a method 8.4 Monitor the run • Make appropriate selections under the tabs described in the table below. Customization Curves Select curves to be displayed. Curve style and color Customize the appearance of the displayed curves. X-axis Select base unit (time or volume) and set X-axis scale. Y-axis Select which Y-axes to display and set Y-axis scale for the different curves.
  • Page 109 8 Run a method 8.4 Monitor the run Customize Run Log The Run Log pane displays all registered actions during the run. Scroll up to see the entire log. To change what items to display in the pane: • Click the Customize icon to open the Customize dialog. •...
  • Page 110 8 Run a method 8.4 Monitor the run Flow scheme The Flow Scheme pane displays the current flow path during the run. Color indication is applied, as shown in the table below. Real-time data from monitors are also displayed in the flow scheme. See illustration below. Color Indication Green...

  • Page 111: After Run Procedures

    8 Run a method 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 the columns should be cleaned between the runs. This will prevent, for example, sample contamination, protein precipitation and column clogging.
  • Page 112 8 Run a method 8.5 After run procedures Column cleaning After a method run is completed, perform the following: • 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: •...
  • Page 113 8 Run a method 8.5 After run procedures Step Action Prepare at least 10 ml storage solution by mixing equal volumes of a standard buffer pH 4 and a 1 M Potassium Nitrate (KNO ) solution. Fill a syringe with approximately 10 ml of the storage solution. Connect the syringe to the pH valve port Cal, and inject the storage solution.
  • Page 114 8 Run a method 8.5 After run procedures If you want to... then... exit UNICORN select File:Exit UNICORN. Result: All open UNICORN modules close. If an edited method or result is open and not saved when you try to exit Note: or log off UNICORN, you will see a warning.

  • Page 115: Evaluate And Print The Results

    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...

  • Page 116: View The Results

    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.
  • Page 117 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. Customize Chromatogram To change the chromatogram properties: • Click the Customize icon to open the Customize dialog. •...
  • Page 118 9 Evaluate and print the results 9.1 View the results 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.
  • Page 119 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.

  • Page 120: Peak Integration

    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.
  • Page 121 9 Evaluate and print the results 9.2 Peak integration Step Action In the Peak Integrate dialog: Select which curve to integrate (e.g., UV1_280 for proteins). • Select a target peak table for storage of the result. • Select Calculate baseline from the Baseline list. •...
  • Page 122 9 Evaluate and print the results 9.2 Peak integration Illustration of Chromatogram and Peak Table The illustration below shows a Chromatogram pane including a Peak Table. Getting Started with ÄKTAavant and UNICORN 6 28-9440-69 AB...
  • Page 123 9 Evaluate and print the results 9.2 Peak 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. • Click the Customize icon to open the Customize dialog. •...
  • Page 124 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.

  • Page 125: Print The Results

    9 Evaluate and print the results 9.3 Print the results 9.3 Print the results Introduction This section describes how to print a chromatogram and a standard format report. UNICORN uses the printers and printer settings that are installed on your computer. Customize the view Before printing, make sure that the view is adjusted to show what is needed in the report/printout.
  • Page 126 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 Open the Evaluation module and open a result. Click the Print icon.
  • Page 127 9 Evaluate and print the results 9.3 Print the results Step Action Are you satisfied with the layout? If Yes, continue to step 5. • If No, select File:Exit to return to the Print Chromatograms dialog and • choose another print format. In the Customize Report window, click the Print icon.
  • Page 128 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 Open the Evaluation module and open a result. Click the Report icon. Result: The Generate Report dialog opens. In the Generate Report dialog: Select any of the predefined formats from the list.
  • Page 129 9 Evaluate and print the results 9.3 Print the results Step Action In the Customize Report dialog, click the Print icon. Result: The Print dialog opens. In the Print dialog, select Printer from the list and click OK. Result: Your report is printed. Getting Started with ÄKTAavant and UNICORN 6 28-9440-69 AB...

  • Page 130: Maintenance

    10 Maintenance 10 Maintenance Introduction This chapter provides schedules for preventive maintenance that should be performed by the user of ÄKTAavant. Regular maintenance is essential for reliable function and results. Refer to ÄKTAavant and UNICORN 6 User Manual for detailed instructions. WARNING Always use appropriate personal protective equipment during operation and maintenance of ÄKTAavant system.

  • Page 131: Maintenance Program

    10 Maintenance 10.1 Maintenance program 10.1 Maintenance program Introduction This section describes the preventive maintenance to be performed on ÄKTAavant. 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 ÄKTAavant.
  • Page 132 10 Maintenance 10.1 Maintenance program Maintenance when required The following maintenance should be performed by the user of ÄKTAavant when required. Maintenance action Clean the instrument externally Perform System CIP Perform Column CIP Clean the Fraction collector Replace tubing and connectors Storage of pH electrode Clean the pH electrode Clean the conductivity flow cell...
  • Page 134 Sweden 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 www.gelifesciences.com/AKTA...

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