GE AKTA avant User manual

Type
User manual
ÄKTA
â„¢
avant
User Manual
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Table of Contents
71 Introduction ..........................................................................................................
81.1 Important user information .............................................................................................................
101.2 ÄKTA avant overview ..........................................................................................................................
121.3 Associated Documentation .............................................................................................................
152 The ÄKTA avant instrument ...............................................................................
162.1 Overview illustrations .........................................................................................................................
302.2 Runs in a cold environment .............................................................................................................
312.3 Removing the foldable door and pump cover ........................................................................
352.4 Liquid flow path .....................................................................................................................................
372.5 Instrument display ...............................................................................................................................
432.6 Accessories ..............................................................................................................................................
442.6.1 Column holders ...............................................................................................................................
482.6.2 Tubing holders and other accessories ..................................................................................
532.6.3 Extension box ..................................................................................................................................
553 Standard instrument modules ...........................................................................
563.1 Pumps ........................................................................................................................................................
603.2 Mixer ...........................................................................................................................................................
623.3 Valves, overview ....................................................................................................................................
643.4 Inlet valves ...............................................................................................................................................
703.5 Injection valve ........................................................................................................................................
733.6 Column valve ..........................................................................................................................................
763.7 Outlet valve ..............................................................................................................................................
783.8 Pressure monitors ................................................................................................................................
803.9 pH valve and pH monitor ..................................................................................................................
833.10 Built-in fraction collector ...................................................................................................................
843.10.1 Function .............................................................................................................................................
863.10.2 Illustrations of the built-in fraction collector ......................................................................
893.10.3 Cassettes, cassette tray and racks .........................................................................................
963.11 UV monitor ...............................................................................................................................................
983.12 Conductivity monitor ..........................................................................................................................
1004 Optional instrument modules ............................................................................
1014.1 System extension overview .............................................................................................................
1024.2 Installation of optional modules ....................................................................................................
1104.3 Extra inlet valves ...................................................................................................................................
1144.4 Second Column valve .........................................................................................................................
1174.5 Extra Outlet valves ...............................................................................................................................
1204.6 External air sensors .............................................................................................................................
1244.7 Loop valve ................................................................................................................................................
1284.8 Versatile valve ........................................................................................................................................
1304.9 Fraction collector F9-R .......................................................................................................................
1374.10 Second Conductivity monitor ..........................................................................................................
ÄKTA avant User Manual 29035184 AE 3
Table of Contents
1394.11 Second UV monitor ..............................................................................................................................
1434.12 I/O-box .......................................................................................................................................................
1444.12.1 Overview of the I/O-box ..............................................................................................................
1464.12.2 Analog connector and signals ..................................................................................................
1484.12.3 Digital connector and signals ...................................................................................................
1504.12.4 Connect external equipment to the I/O-box ......................................................................
1534.13 General system settings ....................................................................................................................
1555 Operation ..............................................................................................................
1565.1 Before you prepare the system ......................................................................................................
1575.2 Prepare the flow path .........................................................................................................................
1625.3 Start UNICORN and connect to the system .............................................................................
1655.4 Prime inlets and purge pump heads ...........................................................................................
1665.4.1 Prime buffer inlets and purge system pumps ...................................................................
1735.4.2 Prime sample inlets and purge Sample Pump ..................................................................
1785.4.3 Prime Q inlets ...................................................................................................................................
1825.5 Connect a column ................................................................................................................................
1875.6 Set pressure alarms .............................................................................................................................
1895.7 Sample application ..............................................................................................................................
1915.7.1 Sample application using direct injection into the column .........................................
1945.7.2 Sample application using a Superloop ................................................................................
1995.7.3 Sample application using a sample loop ............................................................................
2025.8 Fractionation ..........................................................................................................................................
2035.8.1 Prepare the built-in fraction collector ...................................................................................
2105.8.2 Prepare Fraction collector F9-R ...............................................................................................
2185.8.3 Fractionation overview ................................................................................................................
2255.9 Create a method and perform a run ...........................................................................................
2265.9.1 Create a method .............................................................................................................................
2305.9.2 Perform a run ..................................................................................................................................
2335.9.3 Monitor the run ...............................................................................................................................
2365.9.4 After run procedures ....................................................................................................................
2396 Performance tests ...............................................................................................
2406.1 General performance test actions ...............................................................................................
2436.2 Air sensor tests ......................................................................................................................................
2456.3 Built-in Fraction collector test .........................................................................................................
2506.4 Fraction collector F9-R test ..............................................................................................................
2536.5 Quaternary valve test .........................................................................................................................
2566.6 System test ..............................................................................................................................................
2606.7 UV monitor U9-L test ..........................................................................................................................
2627 Maintenance .........................................................................................................
2637.1 Maintenance Manager .......................................................................................................................
2687.2 Maintenance program .......................................................................................................................
2727.3 Weekly maintenance ..........................................................................................................................
2737.3.1 Change pump rinsing solution .................................................................................................
2757.3.2 Replace the inline filter ................................................................................................................
2767.3.3 Clean built-in fraction collector sensors ...............................................................................
4 ÄKTA avant User Manual 29035184 AE
Table of Contents
2787.4 Monthly maintenance ........................................................................................................................
2797.4.1 Check the flow restrictor .............................................................................................................
2817.4.2 Check the function of the pump flow restrictors ..............................................................
2827.5 Semiannual maintenance ................................................................................................................
2837.5.1 Clean the UV flow cell ..................................................................................................................
2867.5.2 Replace the pH electrode ............................................................................................................
2877.6 Maintenance when required ...........................................................................................................
2887.6.1 Clean the instrument externally ..............................................................................................
2897.6.2 Perform System CIP ......................................................................................................................
2977.6.3 Perform Column CIP .....................................................................................................................
3017.6.4 Clean built-in fraction collector ................................................................................................
3047.6.5 Clean Fraction collector F9-R ....................................................................................................
3057.6.6 Storage of the pH electrode ......................................................................................................
3077.6.7 Clean the pH electrode ................................................................................................................
3117.6.8 Regenerate dried out pH electrode ........................................................................................
3127.6.9 Clean pump head check valves ...............................................................................................
3157.6.10 Clean pump flow restrictors ......................................................................................................
3177.6.11 Wipe off excess oil from the pump head .............................................................................
3187.6.12 Clean the Conductivity flow cell ...............................................................................................
3207.7 Calibration procedures ......................................................................................................................
3217.7.1 Calibrate the pH monitor ............................................................................................................
3237.7.2 Calibrate the pressure monitors ..............................................................................................
3277.7.3 Calibrate the Conductivity Monitor ........................................................................................
3317.7.4 Calibrate the UV monitors ..........................................................................................................
3357.8 Replacement procedures ..................................................................................................................
3367.8.1 Replace tubing and connectors ...............................................................................................
3387.8.2 Replace the Mixer ..........................................................................................................................
3407.8.3 Replace the O-ring inside the Mixer .......................................................................................
3427.8.4 Replace the UV monitor U9-M flow cell ...............................................................................
3457.8.5 Replace UV monitor U9-L flow cell .........................................................................................
3477.8.6 Replace Flow restrictor ................................................................................................................
3487.8.7 Replace the inlet filters ................................................................................................................
3497.8.8 Replace pump head check valves ..........................................................................................
3527.8.9 Replace pump rinsing system tubing ....................................................................................
3547.8.10 Replace a valve module ..............................................................................................................
3577.8.11 Replace pump pistons, piston seals, O-rings and rinse membranes ......................
3588 Troubleshooting ...................................................................................................
3598.1 Introduction to troubleshooting .....................................................................................................
3618.2 Troubleshooting: General Checklist .............................................................................................
3638.3 Troubleshooting: Monitors ................................................................................................................
3818.4 Troubleshooting: Valves .....................................................................................................................
3848.5 Troubleshooting: Fraction collector .............................................................................................
3978.6 Troubleshooting: Pumps ....................................................................................................................
4048.7 Troubleshooting: Other components ..........................................................................................
4138.8 Troubleshooting: Instrument communication ........................................................................
4158.9 Error codes ...............................................................................................................................................
ÄKTA avant User Manual 29035184 AE 5
Table of Contents
4219 Reference information ........................................................................................
4229.1 System specifications .........................................................................................................................
4249.2 Module specifications .........................................................................................................................
4329.3 Tubing and connectors ......................................................................................................................
4409.4 Chemical resistance guide ...............................................................................................................
4419.4.1 General information about biocompatibility and chemical resistance ..................
4439.4.2 Chemical resistance specifications ........................................................................................
4479.5 Wetted materials ..................................................................................................................................
4499.6 Predefined methods and phases ..................................................................................................
4599.7 System settings .....................................................................................................................................
4609.7.1 System settings - UV .....................................................................................................................
4619.7.2 System settings - Conductivity .................................................................................................
4639.7.3 System settings - pH .....................................................................................................................
4649.7.4 System settings - Pumps and pressures ..............................................................................
4669.7.5 System settings - Air sensor ......................................................................................................
4679.7.6 System settings - I/O-box ...........................................................................................................
4689.7.7 System settings - Fraction collection .....................................................................................
4719.7.8 System settings - Watch parameters ...................................................................................
4729.7.9 System settings - Advanced ......................................................................................................
4749.7.10 System settings - Data collection ............................................................................................
4759.8 Using Manual instructions ................................................................................................................
4769.8.1 Manual instructions - Pumps and pressures .....................................................................
4799.8.2 Manual instructions - Flow path .............................................................................................
4819.8.3 Manual instructions - Monitors ................................................................................................
4839.8.4 Manual instructions - Fraction collection ............................................................................
4879.8.5 Manual instructions - I/O-box ...................................................................................................
4889.8.6 Manual instructions - Alarms ....................................................................................................
4929.8.7 Manual instructions - Advanced ..............................................................................................
4959.8.8 Manual instructions - Watch parameters ...........................................................................
4969.8.9 Manual instructions - Other .......................................................................................................
4979.9 Available Run data ...............................................................................................................................
5019.10 Available Curves ....................................................................................................................................
5049.11 Injection volumes and peak broadening ...................................................................................
5059.12 Delay volumes ........................................................................................................................................
5099.13 Component volumes ...........................................................................................................................
5129.14 Pressure control ....................................................................................................................................
5159.15 Check and change the Node ID of a module ..........................................................................
51910 Ordering information ..........................................................................................
530Index .......................................................................................................................
6 ÄKTA avant User Manual 29035184 AE
Table of Contents
1 Introduction
Purpose of the User Manual
The User Manual provides you with instructions and information to run the ÄKTA avant
system. It also includes relevant guidance for practical handling and maintenance of
instrument components.
In this chapter
This chapter contains the following sections:
See pageSection
81.1 Important user information
101.2 ÄKTA avant overview
121.3 Associated Documentation
ÄKTA avant User Manual 29035184 AE 7
1 Introduction
1.1 Important user information
Read this before operating the
product
All users must read the entire Operating Instructions before installing, operating or
maintaining the product.
Always keep the Operating Instructions at hand when operating the product.
Do not operate the product in any other way than described in the user documentation.
If you do, you may be exposed to hazards that can lead to personal injury and you may
cause damage to the equipment.
Intended use of the product
ÄKTA avant is a liquid chromatography system intended for method and process devel-
opment in purification of biomolecules. The system can be used to screen for optimal
choice of columns, media and running parameters to purify selected proteins.
The ÄKTA avant system is intended for research use only, and shall not be used in any
clinical procedures, or for diagnostic procedures.
Prerequisites
In order to follow this manual and use the system in the manner it is intended, it is im-
portant that:
•
You have a general understanding of how the computer and Microsoft® Windows®
work.
•
You understand the concepts of liquid chromatography.
•
You have read and understood the Safety instructions chapter in ÄKTA avant
Operating Instructions.
•
A user account has been created according to the UNICORNâ„¢ Administration and
Technical Manual.
8 ÄKTA avant User Manual 29035184 AE
1 Introduction
1.1 Important user information
Safety notices
This user documentation contains safety notices (WARNING, CAUTION, and NOTICE)
concerning the safe use of the product. See definitions below.
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.
CAUTION
CAUTION indicates a hazardous situation which, if not avoided,
could result in minor or moderate injury. It is important not to pro-
ceed until all stated conditions are met and clearly understood.
NOTICE
NOTICE indicates instructions that must be followed to avoid
damage to the product or other equipment.
Notes and tips
A note is used to indicate information that is important for trouble-free and
optimal use of the product.
Note:
A tip contains useful information that can improve or optimize your procedures.
Tip:
ÄKTA avant User Manual 29035184 AE 9
1 Introduction
1.1 Important user information
1.2 ÄKTA avant overview
Introduction
ÄKTA avant is a preparative liquid chromatography system intended for purification of
proteins as well as other bio-molecules and used for method and process development.
The system is equipped with valves, detectors and fraction collector, integrated in the
flow path, needed for automated control and optimal performance. The system can be
used to screen for columns, media, and running parameters to purify selected proteins
This section gives an overview of the ÄKTA avant instrument and UNICORN software.
For detailed information about UNICORN, see the UNICORN manuals listed in User doc-
umentation on the CD, on page 12. For detailed information about the instrument, see
Chapter 2 The ÄKTA avant instrument, on page 15.
Main features
The main features of ÄKTA avant are listed below.
•
The ÄKTA avant is a complete system including all modules needed for normal method
and process development.
•
The ÄKTA avant system can be easily extended with additional modules, e.g. for extra
samples, buffers, outlets, columns and integration of optional components.
•
The ÄKTA avant instrument is compact with a built-in fraction collector.
•
Cooling of the built-in fraction collector enables secure storage of the purified samples
and unattended runs over night.
•
The built-in Instrument display shows instrument status and method state.
•
ÄKTA avant is controlled by the UNICORN software: a complete package for control,
experimental planning, supervision and evaluation of chromatography instruments
and purification runs.
-
Purification and maintenance methods are easily created using predefined
methods and method phases. Method phases are displayed graphically in a
method outline and phase properties, which makes methods and phases easy
to handle and edit.
-
Run statistics history for individual columns can be saved (e.g., total number of
runs and number of runs since last cleaning procedure).
-
BufferPro (automatic buffer preparation) facilitates screening for optimal buffer
compositions as well as preparation of single buffers.
-
Design of Experiments (DoE) is an integrated part of UNICORN that provides an
organized approach that connects experiments in a rational manner.
10 ÄKTA avant User Manual 29035184 AE
1 Introduction
1.2 ÄKTA avant overview
UNICORN modules overview
UNICORN consists of four modules: Administration, Method Editor, System Control and
Evaluation. The main functions of each module are described in the following table.
Main functionsModule
Perform user and system setup, system log and database
administration.
Administration
Create and edit methods using one or a combination of:Method Editor
•
Predefined methods with built-in application support
•
Drag-and-drop function to build methods with relevant
steps
•
Line-by-line text editing
The interface provides easy viewing and editing of run
properties.
Start, monitor and control runs. The current flow path is
illustrated in the Process Picture, which allows manual
interactions with the system and provides feedback on
run parameters.
System Control
Open results, evaluate runs and create reports.Evaluation
The default Evaluation module includes a user interface
optimized for workflows like quick evaluation, compare
results and work with peaks and fractions.
When working with the modules Administration, Method Editor, System Control and
Evaluation it is possible to access descriptions of the active window by pressing the F1
key. This can be especially helpful when editing methods
ÄKTA avant User Manual 29035184 AE 11
1 Introduction
1.2 ÄKTA avant overview
1.3 Associated Documentation
Introduction
This section describes the user documentation that is delivered with ÄKTA avant.
User documentation on the CD
The user documentation listed in the table below is available in printed or PDF format.
The complete documentation is also available on the User Documentation CD.
Main contentsDocument
Instructions for unpacking the instrument,
and how to lift the instrument onto a bench.
ÄKTA avant Unpacking Instructions
(29101559)
Instructions needed to install, operate and
maintain the system in a safe way.
ÄKTA avant Operating Instructions
(29101556)
Instructions for handling the system. Descrip-
tions of components. Information about how
to run and maintain the system.
ÄKTA avant User Manual
(29035184)
System specification and declaration of ma-
terial conformity.
ÄKTA avant 25 Product
Documentation
(28991000) OR
ÄKTA avant 150 Product
Documentation (28984249)
1
1
The instrument is delivered with the relevant document.
UNICORN user documentation
The user documentation listed in the following table is available from the Help menu in
UNICORN or from the UNICORN Online Help and Documentation software accessed
by pressing the F1 key in any UNICORN module.
Main contentsDocumentation
Descriptions of UNICORN dialog boxes (available from
the Help menu).
UNICORN Help
12 ÄKTA avant User Manual 29035184 AE
1 Introduction
1.3 Associated Documentation
Main contentsDocumentation
•
Video clips showing common workflows in the
Evaluation module.
•
Overview of features of the Evaluation module.
Getting started with
Evaluation
Note:
Available in UNICORN 7.0
and later.
•
Overview and detailed descriptions of the method
creation features in UNICORN.
•
Workflow descriptions for common operations.
UNICORN Method Manual
1
•
Overview and detailed description of network
setup and complete software installation.
•
Administration of UNICORN and the UNICORN
database.
UNICORN Administration
and Technical Manual
1
•
Overview and detailed descriptions of the
Evaluation Classic module in UNICORN.
•
Description of the evaluation algorithms used in
UNICORN.
UNICORN Evaluation
Manual
1
•
Overview and detailed description of the system
control features in UNICORN.
•
Includes general operation, system settings and
instructions on how to perform a run.
UNICORN System Control
Manual
1
1
Current UNICORN version is added to the title of the manual.
Data files, application notes and
user documentation on the web
To order or download data files, application notes or user documentation, see the instruc-
tion below.
ActionStep
Go to www.gelifesciences.com/avant.1
Click Product support.2
Click Related Documents.3
Select to download the chosen literature.4
ÄKTA avant User Manual 29035184 AE 13
1 Introduction
1.3 Associated Documentation
Additional literature
For practical tips on chromatography, refer to ÄKTA Laboratory-scale: Chromatography
Systems Instrument Management Handbook (product code 29010831).
14 ÄKTA avant User Manual 29035184 AE
1 Introduction
1.3 Associated Documentation
2 The ÄKTA avant instrument
About this chapter
This chapter provides an overview of the ÄKTA avant instrument and the standard and
optional instrument modules.
In this chapter
This chapter contains the following sections:
See pageSection
162.1 Overview illustrations
302.2 Runs in a cold environment
312.3 Removing the foldable door and pump cover
352.4 Liquid flow path
372.5 Instrument display
432.6 Accessories
ÄKTA avant User Manual 29035184 AE 15
2 The ÄKTA avant instrument
2.1 Overview illustrations
Introduction
This section provides an overview of the system and its available modules.
Instrument configurations
ÄKTA avant is available with two standard module configurations, one for flow rates up
to 25 ml/min and one for flow rates up to 150 ml/min. In this manual they are referred
to as ÄKTA avant 25 (25 ml/min) and ÄKTA avant 150 (150 ml/min).
Illustration of the main parts of
the instrument
The following illustration shows the location of the main parts of the instrument.
FunctionPartFunctionPart
Buffer tray2Fraction collector1
Wet side4Instrument display3
Pump cover6Foldable door5
16 ÄKTA avant User Manual 29035184 AE
2 The ÄKTA avant instrument
2.1 Overview illustrations
FunctionPartFunctionPart
Swivel foot lock/unlock knob8Holder rails7
Swing out toolbox10Swivel foot9
Power switch11
Illustration of the wet side
modules of the instrument
The following illustration shows the modules of the wet side of the instrument.
1 2 3 4 5 6
7
8
9
1011121314151617
18
19
20
21
FunctionPartFunctionPart
Flow restrictor2Injection valve1
Column valve4pH valve3
Conductivity monitor6UV monitor5
Quaternary valve8Inlet valve B7
Pressure monitor of system
pumps
10System pump B9
Inlet valve A12System pump flow restrictor11
ÄKTA avant User Manual 29035184 AE 17
2 The ÄKTA avant instrument
2.1 Overview illustrations
FunctionPartFunctionPart
Mixer14System pump A13
Pressure monitor of sample
pump
16Sample pump flow restrictor15
Pump rinsing solution tube18Sample pump17
Outlet valve20Sample inlet valve19
Holder rails21
Standard modules
ÄKTA avant is delivered with the modules listed in the following table.
Label inModule
ÄKTA avant 150ÄKTA avant 25
P9H AP9 ASystem pump A
P9H BP9 BSystem pump B
P9H SP9-SSample pump
R9R9Pressure monitor
M9M9Mixer
V9H-InjV9-InjInjection valve
Q9Q9Quarternary valve
V9H-IAV9-IAInlet valve A
V9H-IBV9-IBInlet valve B
V9H-ISV9-ISSample inlet valve
V9H-CV9-CColumn valve
V9H-pHV9-pHpH valve
V9H-OV9-OOutlet valve
U9-MU9-MUV monitor
C9C9Conductivity monitor
18 ÄKTA avant User Manual 29035184 AE
2 The ÄKTA avant instrument
2.1 Overview illustrations
Label inModule
ÄKTA avant 150ÄKTA avant 25
NANABuilt-in fraction collector
Optional modules
One or more of the modules in the following table may be added to the flow path of
ÄKTA avant.
Label inModule
ÄKTA avant 150ÄKTA avant 25
V9H-A2V9-A2Second Inlet valve A
V9H-B2V9-B2Second Inlet valve B
V9H-IXV9-IXExtra Inlet valve X1
V9H-IXV9-IXExtra Inlet valve X2
V9H-S2V9-S2Second Sample inlet valve
V9H-VV9-VVersatile valve
V9H-LV9-LLoop valve
V9H-C2V9-C2Second Column valve
V9H-O2V9-O2Second Outlet valve
V9H-O3V9-O3Third Outlet valve
L9-1.5L9-1.5External air sensor L9-1.5
L9-1.2L9-1.2External air sensor L9-1.2
E9E9I/O-box
U9-LU9-LSecond UV monitor
C9C9Second Conductivity
monitor
F9-RF9-RSecond Fraction collector
ÄKTA avant User Manual 29035184 AE 19
2 The ÄKTA avant instrument
2.1 Overview illustrations
Illustration convention
In the valve illustrations below, the following convention is used to point out the location
of the ports on the valve head. Loop valve V9-L is used as an example.
3E
3F
V9-L
1
E
2
E
4
E
5
E
5
F
4
F
2
F
1
F
E
F
•
Ports located on the valve head rim are indicated outside
the black ring (e.g., 1E, 2E, etc.).
•
Ports located on the pivot part of the valve head are in-
dicated on the inside of the black ring (e.g., 3E and 3F).
•
Ports located on the valve head front are indicated by
an arrow (e.g., E and F).
Description of standard modules
The following modules are installed in the instrument when delivered.
DescriptionModule
A high precision pump, which delivers buffer in
purification runs.
System pump A P9 A or P9H A
For further information, refer to Section 3.1 Pumps,
on page 56.
20 ÄKTA avant User Manual 29035184 AE
2 The ÄKTA avant instrument
2.1 Overview illustrations
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