Getting Started with ÄKTA avant

Getting Started with ÄKTA avant
GE Healthcare
Life Sciences
Getting Started with
ÄKTA™ avant
Original instructions
Table of Contents
Table of Contents
1
Introduction ..........................................................................................................
1.1
1.2
5
Important user information .............................................................................................................
Regulatory information ......................................................................................................................
6
10
Safety instructions ...............................................................................................
14
2.1
2.2
2.3
2.4
2.5
Safety precautions ...............................................................................................................................
Labels .........................................................................................................................................................
Emergency procedures ......................................................................................................................
Recycling information .........................................................................................................................
Declaration of Hazardous Substances (DoHS) ........................................................................
15
26
29
32
33
3
Start up .................................................................................................................
36
4
The instrument and software ............................................................................
40
4.1
ÄKTA avant instrument overview ..................................................................................................
4.2
UNICORN software overview ..........................................................................................................
4.3
UNICORN software modules ...........................................................................................................
4.3.1
Method Editor module ......................................................................................................................
4.3.2
System Control module ...................................................................................................................
4.3.3
Evaluation module .............................................................................................................................
41
44
47
48
51
53
Create a method ..................................................................................................
55
5.1
Guide to method creation .................................................................................................................
5.1.1
Create and edit methods ................................................................................................................
5.1.2
Predefined methods ..........................................................................................................................
5.1.3
Predefined phases .............................................................................................................................
5.2
Print a method .......................................................................................................................................
56
57
65
67
68
Prepare the system for a run .............................................................................
70
2
5
6
6.1
6.2
6.3
6.4
6.5
6.6
6.7
7
Before you prepare the system ......................................................................................................
Prepare the flow path .........................................................................................................................
Prime buffer inlets and purge system pumps .........................................................................
Connect a column ................................................................................................................................
Calibrate the pH monitor ..................................................................................................................
Prepare the Fraction collector ........................................................................................................
Prepare for a run in a cold cabinet ...............................................................................................
71
73
81
89
94
96
102
Run a method ........................................................................................................ 104
7.1
7.2
7.3
7.4
7.5
Before you start .....................................................................................................................................
Applying the sample ............................................................................................................................
Start a method run ..............................................................................................................................
Monitor the run ......................................................................................................................................
After run procedures ...........................................................................................................................
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105
108
111
116
121
3
Table of Contents
8
Evaluate and print the results ........................................................................... 125
8.1
8.2
8.3
9
4
View the results .....................................................................................................................................
Peak integration ....................................................................................................................................
Print the results ......................................................................................................................................
126
130
135
Maintenance ......................................................................................................... 140
Getting Started with ÄKTA avant 29-0351-83 AA
1 Introduction
1
Introduction
About this chapter
This chapter contains important user information, description of safety notices, regulatory information and a general description of the intended use of ÄKTA avant.
In this chapter
This chapter contains the following sections:
Section
1.1 Important user information
1.2 Regulatory information
Getting Started with ÄKTA avant 29-0351-83 AA
See page
6
10
5
1 Introduction
1.1 Important user information
1.1
Important user information
Read this before using the ÄKTA
avant system
All users must read the entire ÄKTA avant Installation Guide and Getting Started with
ÄKTA avant, before installing, operating, or maintaining the instrument.
Do not operate the ÄKTA avant system in any other way than described in the user
documentation. Otherwise, you may be exposed to hazards that can lead to personal
injury, and you may cause damage to the equipment.
Purpose of Getting Started
The purpose of this guide is to present a quick and easy guide to the system for a user
with limited or no experience of UNICORN™ software and ÄKTA avant instrument. The
work flow is presented as practical instructions on how to operate the software and the
instrument. The instructions form a basic framework that you can expand on by reading
selected parts in the other manuals. This Getting Started guide includes the following
topics:
•
Basic features of ÄKTA avant and UNICORN
•
Create methods
•
Prepare the system for runs
•
Perform runs
•
Make simple evaluations
•
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:
•
6
you have a general understanding of how the computer and Windows™ work.
Getting Started with ÄKTA avant 29-0351-83 AA
1 Introduction
1.1 Important user information
•
you understand the concepts of liquid chromatography.
•
you have read and understood the Safety instructions chapter in the user documentation.
•
the instrument and software are installed, configured and calibrated according to
ÄKTA avant Installation Guide.
•
a user account has been created according to UNICORN 6 Administration and
Technical Manual.
Pushing the limits beyond
Getting Started
Users who are familiar with the instrument and software and want to learn more about
the system’s advanced features should refer to the list below.
To find out more
about...
please read..
installation
ÄKTA avant Installation Guide
administration of
databases
UNICORN 6 Administration and Technical Manual
instrument modules and
functions
ÄKTA avant User Manual
calibration
ÄKTA avant User Manual
predefined methods
UNICORN 6 Method Manual
column handling
UNICORN 6 Method Manual
manual method editing
UNICORN 6 Method Manual
BufferPro–automatic
buffer preparation
UNICORN 6 Method Manual
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 ÄKTA avant 29-0351-83 AA
7
1 Introduction
1.1 Important user information
Intended use
ÄKTA avant is a liquid chromatography system intended for process development. The
system can be used to screen for optimal choice of columns, media and running 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 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.
Notes and tips
Note:
8
A Note is used to indicate information that is important for trouble-free and
optimal use of the product.
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1 Introduction
1.1 Important user information
Tip:
A tip contains useful information that can improve or optimize your procedures.
Typographical conventions
Software items are identified in the text by bold italic text. A colon separates menu levels,
thus File:Open refers to the Open command in the File menu.
Hardware controls, indicators and connections are identified in the text by bold text (e.g.,
Power switch).
Text entries that UNICORN generates or that the user must type are represented by a
monotype typeface (e.g., \Program Files\GE Healthcare\UNICORN\bin\UNICORN Instrument Server.exe.config).
Getting Started with ÄKTA avant 29-0351-83 AA
9
1 Introduction
1.2 Regulatory information
1.2
Regulatory information
Introduction
This section describes the directives and standards that are fulfilled by ÄKTA avant.
Manufacturing information
The table below summarizes the required manufacturing information. For further information, see the EC Declaration of Conformity document.
Requirement
Content
Name and address of
manufacturer
GE Healthcare Bio-Sciences AB, Björkgatan 30, SE-751
84 Uppsala, Sweden
CE conformity
This product complies with the European directives listed in the table below, by fulfilling
the corresponding harmonized standards. For further information, see the EC Declaration
of Conformity document.
Directive
Title
2006/42/EC
Machinery Directive (MD)
2006/95/EC
Low Voltage Directive (LVD)
2004/108/EC
ElectroMagnetic Compatibility (EMC) Directive
1999/5/EC
Radio Equipment and Telecommunications Terminal
Equipment (R&TTE) Directive
CE marking
The CE marking and the corresponding EC Declaration of Conformity, is valid for the instrument when it is:
•
10
used as a stand-alone unit, or
Getting Started with ÄKTA avant 29-0351-83 AA
1 Introduction
1.2 Regulatory information
•
connected to other GE Healthcare instruments, or
•
connected to other products recommended or described in the user documentation,
and
•
used in the same state as it was delivered from GE Healthcare, except for alterations
described in the user documentation.
International standards
This product fulfills the requirements of the following standards:
Standard
Description
Notes
EN ISO 12100
Safety of machinery. General principles for design. Risk assessment and
risk reduction.
EN ISO standard is harmonized with EU directive 2006/42/EC
EN 61010-1, IEC
61010-1, UL
61010-1, CAN/CSA
C22.2 No. 61010-1
Safety requirements for electrical
equipment for measurement, control, and laboratory use.
EN standard is harmonized with EU directive
2006/95/EC
EN 61326-1, IEC
61326-1
Electrical equipment for measurement, control and laboratory use EMC requirements
EN standard is harmonized with EU directive
2004/108/EC
EN 301 489-1, EN
301 489-3
Electromagnetic compatibility and
Radio spectrum Matters (ERM);
ElectroMagnetic Compatibility (EMC)
standard for radio equipment and
services.
EN standard is harmonized with EU directives
1999/5/EC and
2004/108/EC.
EN 300 330-2
Electromagnetic compatibility and
Radio spectrum Matters (ERM); Short
Range Devices (SRD); Radio equipment in the frequency range 9 kHz
to 25 MHz and inductive loop systems in the frequency range 9 kHz
to 30 MHz.
EN standard is harmonized with EU directive
1999/5/EC.
(Emission according to CISPR 11,
Group 1, class A)
Getting Started with ÄKTA avant 29-0351-83 AA
11
1 Introduction
1.2 Regulatory information
FCC statement
The ÄKTA avant instrument, excluding transmitted radio frequency energy from the
Mixer M9 and UV detector U9-D, complies with FCC 47 CFR Part 15b (Federal Communications Commission (FCC) title 47 of the Code of Federal Regulations (CFR), Part 15b,
Radio Frequency Devices).
Note:
This equipment has been tested and found to comply with the limits for a Class
A digital device, pursuant to part 15 of the FCC Rules. These limits are designed
to provide reasonable protection against harmful interference when the
equipment is operated in a commercial environment. This equipment generates,
uses and can radiate radio frequency energy and, if not installed and used in
accordance with the instruction manual, may cause harmful interference to
radio communications. Operation of this equipment in a residential area is
likely to cause harmful interference in which case the user will be required to
correct the interference at his own expense.
The modules Mixer M9 and UV detector U9-D, comply with FCC 47 CFR Part 15c.
Note:
This device complies with Part 15c rules. Operation is subject to the following
two conditions:
•
This device may not cause harmful interference, and
•
This device must accept any interference received, including interference
that may cause undesired operation.
The user is cautioned that any changes or modifications not expressly approved
by the manufacturer could void the user’s authority to operate the equipment.
Software declaration of
conformity
UNICORN 6 is technically compatible with all relevant sections of FDA 21 CFR Part 11.
A part 11-system assessment checklist is available on request through the local GEHC
representative.
Regulatory compliance of
connected equipment
Any equipment connected to ÄKTA avant should meet the safety requirements of EN
61010-1/IEC 61010-1, or other relevant harmonized standards. Within EU, connected
equipment must be CE marked.
12
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1 Introduction
1.2 Regulatory information
Environmental Conformity
Regulation
Title
2011/65/EU
Restriction of Hazardous Substances (RoHS) Directive
2002/96/EC
Waste Electrical and Electronic Equipment (WEEE) Directive
Regulation (EC) No
1907/2006
Registration, Evaluation, Authorization and restriction
of CHemicals (REACH)
ACPEIP
Administration on the Control of Pollution Caused by
Electronic Information Products, China Restriction of
Hazardous Substances (RoHS)
Getting Started with ÄKTA avant 29-0351-83 AA
13
2 Safety instructions
2
Safety instructions
About this chapter
This chapter contains instructions of how to handle the ÄKTA avant instrument in a safe
way.
In this chapter
This chapter contains the following sections:
Section
14
See page
2.1 Safety precautions
15
2.2 Labels
26
2.3 Emergency procedures
29
2.4 Recycling information
32
2.5 Declaration of Hazardous Substances (DoHS)
33
Getting Started with ÄKTA avant 29-0351-83 AA
2 Safety instructions
2.1 Safety precautions
2.1
Safety precautions
Introduction
The ÄKTA avant instrument is powered by mains voltage and handles pressurized liquids
that may be hazardous. Before installing, operating or maintaining the system, you must
be aware of the hazards described in this manual. Follow the instructions provided to
avoid personal injuries or damage to the equipment.
The safety precautions in this section are grouped into the following categories:
•
General precautions
•
Flammable liquids
•
Personal protection
•
Installing and moving the instrument
•
System operation
•
Maintenance
Getting Started with ÄKTA avant 29-0351-83 AA
15
2 Safety instructions
2.1 Safety precautions
General precautions
WARNING
Always follow these General precautions to avoid injury when using
the ÄKTA avant instrument.
•
Do not operate the ÄKTA avant instrument in any other way
than described in the ÄKTA avant and UNICORN manuals.
•
Operation and user maintenance of the ÄKTA avant instrument
should be performed by properly trained personnel only.
•
To avoid exposing the column to excessive pressure, make sure
that the pressure limit is set to the specified maximum pressure
of the column. Before connecting a column to the ÄKTA avant
instrument, read the instructions for use of the column.
•
Do not use any accessories not supplied or recommended by
GE Healthcare.
•
Do not use the ÄKTA avant instrument if it is not working
properly, or if it has suffered any damage, for example:
-
damage to the power cord or its plug
-
damage caused by dropping the equipment
-
damage caused by splashing liquid onto it
NOTICE
Avoid condensation by letting the instrument equilibrate to ambient
temperature.
16
Getting Started with ÄKTA avant 29-0351-83 AA
2 Safety instructions
2.1 Safety precautions
Using flammable liquids
WARNING
When using flammable liquids with the ÄKTA avant instrument,
follow these precautions to avoid any risk of fire or explosion.
•
Fire Hazard. Before starting the system, make sure that there
are no unintentional leakage in the instrument or tubing.
•
Explosion hazard. To avoid building up an explosive atmosphere when using flammable liquids, make sure that the room
ventilation meets the local requirements.
•
Fraction collector. Do not fractionate flammable liquids. When
running RPC methods, or other procedures using organic solvents, collect fractions through the Outlet valve.
•
RPC runs with 100% acetonitrile in ÄKTA avant 25. Always
replace the green PEEK tubing between the used system pump
and the pump pressure monitor with orange PEEK tubing, i.d.
0.5 mm, before running RPC with 100% acetonitrile. Set the
System pressure alarm to 10 MPa.
•
RPC runs with 100% acetonitrile in ÄKTA avant 150. Always
replace the beige PEEK tubing between the used system pump
and the pump pressure monitor with orange PEEK tubing, i.d.
0.5 mm, before running RPC with 100% acetonitrile.
Getting Started with ÄKTA avant 29-0351-83 AA
17
2 Safety instructions
2.1 Safety precautions
Personal protection
WARNING
To avoid hazardous situations when working with the ÄKTA avant
system, take the following measures for personal protection.
•
Always use appropriate personal protective equipment during
operation and maintenance of ÄKTA avant 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.
•
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.
•
High pressure. The ÄKTA avant instrument operates under
high pressure. Wear protective glasses at all times.
CAUTION
To avoid hazardous situations when working with the ÄKTA avant
system, take the following measures for personal protection.
18
•
Always use appropriate personal protective equipment when
decommissioning the equipment.
•
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.
•
Cut injuries. The tubing cutter is very sharp and must be handled with care to avoid injuries.
Getting Started with ÄKTA avant 29-0351-83 AA
2 Safety instructions
2.1 Safety precautions
Installing and moving the
instrument
WARNING
To avoid personal injury when installing or moving the ÄKTA avant
instrument, follow the instructions below.
•
Heavy object. The ÄKTA avant instrument weighs about 116
kg. Use proper lifting equipment, or use four or more persons
when moving the instrument. All lifting and moving must be
performed in accordance with local regulations.
•
Moving the instrument horizontally. Three persons are recommended when moving the instrument horizontally.
•
Supply voltage. Make sure that the supply voltage at the wall
outlet corresponds to the marking on the instrument, before
connecting the power cord.
•
Protective ground. The ÄKTA avant instrument must always
be connected to a grounded power outlet.
•
Power cord. Only use grounded power cords delivered or approved by GE Healthcare.
•
Access to power switch and power cord. Do not block the rear
and side panel of the instrument. The Power switch must always
be easy to access. The power cord must always be easy to
disconnect.
•
Installing the computer. The computer should be installed and
used according to the instructions provided by the manufacturer of the computer.
CAUTION
Make sure that the waste vessels will hold all the produced volume
of the run. For ÄKTA avant 25, a suitable waste vessel should typically have a volume of 2 to 10 liters. For ÄKTA avant 150, a waste
vessel should have a volume of 40 liters.
Getting Started with ÄKTA avant 29-0351-83 AA
19
2 Safety instructions
2.1 Safety precautions
NOTICE
To avoid damage to ÄKTA avant or other equipment when installing
or moving the instrument, follow the instructions below.
20
•
The maximum level of the waste vessel for the waste tubing
from the valves must be lower than 30 cm above the lab bench.
•
The maximum level of the waste vessel for the waste tubing
from the Fraction collector and the Buffer tray must be lower
than the bench height.
•
Vents on the ÄKTA avant 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 ÄKTA avant instrument before an instrument 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.
Getting Started with ÄKTA avant 29-0351-83 AA
2 Safety instructions
2.1 Safety precautions
System operation
WARNING
To avoid personal injury when operating the ÄKTA avant system,
follow the instructions below.
•
Rotating the instrument. Make sure that there is always at
least 20 cm of free space around the ÄKTA avant instrument
to allow for sufficient ventilation and rotation on the swivel foot.
When rotating the instrument, take care not to stretch or
squeeze tubing or cables. A disconnected cable may cause
power interruption or network interruption. Stretched tubing
may cause bottles to fall, resulting in liquid spillage and shattered glass. Squeezed tubing may cause increase in pressure,
or block liquid flow. To avoid the risk of knocking over bottles,
always place bottles on the buffer tray, and close the doors
before rotating the instrument.
•
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.
•
Electrical shock hazard after spillage. If there is a risk that
large volumes of spilled liquid may penetrate the casing of the
ÄKTA avant instrument, immediately switch off the instrument,
disconnect the power cord, and contact an authorized service
engineer.
Getting Started with ÄKTA avant 29-0351-83 AA
21
2 Safety instructions
2.1 Safety precautions
WARNING
22
•
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.
•
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.
•
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.
•
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 ÄKTA avant 29-0351-83 AA
2 Safety instructions
2.1 Safety precautions
CAUTION
To avoid hazardous situations when operating the ÄKTA avant
system, follow the instructions below.
•
Risk of explosion. Do not use Mixer chamber 15 ml in ÄKTA
avant 25. The maximum pressure for Mixer chamber 15 ml is
5 MPa.
•
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 Buffer tray. Do not place containers with a
volume of more than 10 liters each on the Buffer tray. The total
allowed weight on the Buffer tray is 40 kg.
•
Max. size of bottles on front panel. Do not fasten bottles with
a volume of more than 1 liter in the front panel rails.
•
Risk of explosion. Do not use Mixer chamber 15 ml in ÄKTA
avant 25. The maximum pressure for Mixer chamber 15 ml is
5 MPa.
Getting Started with ÄKTA avant 29-0351-83 AA
23
2 Safety instructions
2.1 Safety precautions
NOTICE
To avoid damage to ÄKTA avant or other equipment when operating
the instrument, follow the instructions below.
24
•
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.
•
Glass tube splinter. Make sure to set the sample pressure below the max pressure of the Superloop before executing a flow
in the Manual instructions dialog when the Superloop is connected.
•
Avoid condensation. If ÄKTA avant is kept in a cold room, cold
cabinet or similar, keep it switched on in order to avoid condensation.
•
Avoid overheating. If ÄKTA avant is kept in a cold cabinet and
the cold cabinet is switched off, make sure to switch off ÄKTA
avant and keep the cold cabinet open to avoid overheating.
•
Place the computer in room temperature. If the ÄKTA avant
instrument is placed in a cold room, use a cold room compatible
computer or place the computer outside the cold room and
use the Ethernet cable delivered with the instrument to connect
to the computer.
Getting Started with ÄKTA avant 29-0351-83 AA
2 Safety instructions
2.1 Safety precautions
Maintenance
WARNING
To avoid personal injury when performing maintenance on the
ÄKTA avant instrument, follow the instructions below.
•
Electrical shock hazard. All repairs should be done by service
personnel authorized by GE Healthcare. Do not open any covers
or replace parts unless specifically stated in the user documentation.
•
Disconnect power. Always switch off power to the instrument
before replacing any component on the instrument, unless
stated otherwise in the user documentation.
•
Hazardous chemicals during maintenance. When using hazardous chemicals for system or column cleaning, wash the
system or columns with a neutral solution in the last phase or
step.
NOTICE
To avoid damage to ÄKTA avant or other equipment when performing maintenance on the ÄKTA avant instrument, follow the instructions below.
•
Cleaning. Keep the ÄKTA avant instrument dry and clean. Wipe
regularly with a soft damp tissue and, if necessary, a mild
cleaning agent. Let the ÄKTA avant instrument dry completely
before use.
•
Advanced maintenance. Read the instruction carefully before
disassembly of the pump head.
Getting Started with ÄKTA avant 29-0351-83 AA
25
2 Safety instructions
2.2 Labels
2.2
Labels
Introduction
This section describes the safety labels and labels concerning hazardous substances
that are attached to the ÄKTA avant instrument. For information about marking of the
computer equipment, refer to the manufacturer’s instructions.
Labels on the ÄKTA avant
instrument
The illustrations below show the labels that are attached to the ÄKTA avant instrument.
26
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2 Safety instructions
2.2 Labels
Safety symbols
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!
Do NOT fractionate
flammable liquids.
Fraction collector. Do not fractionate flammable liquids. When running RPC methods, or other procedures
using organic solvents, 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.
This symbol indicates that the system has been certified by a Nationally Recognized Testing Laboratory
(NRTL). An NRTL is an organization that the Occupational Safety and Health Administration (OSHA) has recognized as meeting the legal requirements in USA title
29 of the Code of Federal Regulations (29 CFR) Part
1910.7.
Getting Started with ÄKTA avant 29-0351-83 AA
27
2 Safety instructions
2.2 Labels
Labels concerning hazardous
substances
The following symbols on the labels concern hazardous substances:
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.
This symbol indicates that the product might contain
hazardous materials in excess of the limits established by
the Chinese standard SJ/T11363-2006 Requirements for
Concentration Limits for Certain Hazardous Substances in
Electronic Information Product.
28
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2 Safety instructions
2.3 Emergency procedures
2.3
Emergency procedures
Introduction
This section describes how to perform an emergency shutdown of the ÄKTA avant system.
This section also describes the results in the event of power failure or network interruption.
Emergency shutdown
In an emergency situation, follow the steps below to stop the run:
Step
Action
1
Press the Pause button on the instrument display. This will stop all pumps
in the instrument.
To pause the run from UNICORN, click the Pause icon:
Getting Started with ÄKTA avant 29-0351-83 AA
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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.
30
Power failure to...
will result in...
ÄKTA avant instrument
•
The run is interrupted immediately
•
The data collected up to the time of the power
failure is available in UNICORN.
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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
power failure can be recovered
Note:
The UNICORN client may close down during a
temporary overload of the processor. This may
appear as a computer failure. The run continues
and you can restart the UNICORN client to regain
control.
Uninterruptible power supply
(UPS)
A UPS can prevent data loss during a power failure, and allow time for a controlled shutdown of ÄKTA avant.
For UPS power requirements, see the system specifications in the ÄKTA avant User
Manual. Remember to also take into account the specifications for the computer and
monitor. Refer to the manufacturers' documentation.
Getting Started with ÄKTA avant 29-0351-83 AA
31
2 Safety instructions
2.4 Recycling information
2.4
Recycling information
Introduction
This section describes the procedures for disposal and recycling of ÄKTA avant
Decommissioning and disposal
of the equipment
When taking ÄKTA avant 25 and 150 out of service:
•
The equipment must be decontaminated (performed by an Administrator).
•
The components must be separated and recycled according to national and local
environmental regulations
CAUTION
Always use appropriate personal protective equipment when
decommissioning the equipment.
Disposal of electrical
components
Waste comprising electrical and electronic equipment must not be disposed of as unsorted municipal waste and must be collected separately. Please contact an authorized
representative of the manufacturer for information concerning the decommissioning of
equipment.
32
Getting Started with ÄKTA avant 29-0351-83 AA
2 Safety instructions
2.5 Declaration of Hazardous Substances (DoHS)
2.5
Declaration of Hazardous Substances (DoHS)
Introduction
The following product pollution control information is provided according to SJ/T113642006 Marking for Control of Pollution caused by Electronic Information Products.
根据SJ/T11364-2006《电子信息产品污染控制标识要求》特提供如下有关污染 控制
方面的信息
Symbols used in pollution control
label
电子信息产品污染控制标志说明
Label
Meaning
This symbol indicates the product contains hazardous materials in excess of the limits established by the Chinese standard SJ/T11363-2006
Requirements for Concentration Limits for Certain Hazardous Substances in Electronic Information Products. The number in the symbol
is the Environment-friendly Use Period (EFUP), which indicates the period
during which the toxic or hazardous substances or elements contained
in electronic information products will not leak or mutate under normal
operating conditions so that the use of such electronic information
products will not result in any severe environmental pollution, any
bodily injury or damage to any assets. The unit of the period is “Year”.
In order to maintain the declared EFUP, the product shall be operated
normally according to the instructions and environmental conditions
as defined in the product manual, and periodic maintenance schedules
specified in Product Maintenance Procedures shall be followed strictly.
Consumables or certain parts may have their own label with an EFUP
value less than the product. Periodic replacement of those consumables
or parts to maintain the declared EFUP shall be done in accordance
with the Product Maintenance Procedures.
This product must not be disposed of as unsorted municipal waste,
and must be collected separately and handled properly after decommissioning.
Getting Started with ÄKTA avant 29-0351-83 AA
33
2 Safety instructions
2.5 Declaration of Hazardous Substances (DoHS)
Label
Meaning
该标志表明本产品含有超过SJ/T11363-2006《电子信息产品中有毒
有害物质的限 量要求》中限量的有毒有害物质。标志中的数字为本
产品的环保使用期,表明本 产品在正常使用的条件下,有毒有害物
质不会发生外泄或突变,用户使用本产品 不会对环境造成严重污染
或对其人身、财产造成严重损害的期限。单位为年。
为保证所申明的环保使用期限,应按产品手册中所规定的环境条件
和方法进行正 常使用,并严格遵守产品维修手册中规定的期维修和
保养要求。
产品中的消耗件和某些零部件可能有其单独的环保使用期限标志,
并且其环保使 用期限有可能比整个产品本身的环保使用期限短。应
到期按产品维修程序更换那 些消耗件和零部件,以保证所申明的整
个产品的环保使用期限。
本产品在使用寿命结束时不可作为普通生活垃圾处理,应被单独收
集妥善处理
List of hazardous substances and
their concentrations
产品中有毒有害物质或元素的名称及含量
Indication for each major part if substance exceeds limit
Value
Meaning
O
Indicates that this toxic or hazardous substance contained in all of the
homogeneous materials for this part is below the limit requirement in
SJ/T11363-2006.
表示该有毒有害物质在该部件所有均质材料中的含量均在SJ/T113632006 标准规定的限量要 求以下
X
Indicates that this toxic or hazardous substance contained in at least
one of the homogeneous materials used for this part is above the limit
requirement in SJ/T11363-2006.
•
Data listed in the table represents best information available at the
time of publication
表示该有毒有害物质至少在该部件的某一均质材料中的含量超出
SJ/T11363-2006 标准规定的
限量要求
•
34
此表所列数据为发布时所能获得的最佳信息
Getting Started with ÄKTA avant 29-0351-83 AA
2 Safety instructions
2.5 Declaration of Hazardous Substances (DoHS)
Component
name
Hazardous substance (0/X)
有毒有害物质或元素 (0/X)
部件名称
1
Pb
Hg
Cd
Cr6+
PBB
PBDE
铅
汞
镉
六价铬
多溴联苯
多溴二苯醚
ÄKTA avant 25,
28-9308-42 1
0
0
0
0
0
0
ÄKTA avant 150,
28-9763-371
0
0
0
0
0
0
The product has not been tested as per the Chinese standard SJ/T11363-2006 Requirements
for Concentration Limits for Certain Hazardous Substances in Electronic Information Product.
Getting Started with ÄKTA avant 29-0351-83 AA
35
3 Start up
3
Start up
Introduction
This chapter describes how to start up the system, both the software and the instrument.
It is assumed that your user profile is already created. For information about how to set
up and define users, user groups and access rights, please refer to UNICORN 6
Administration and Technical Manual.
Start the instrument and the
computer
Follow the instructions below to start the instrument and the computer.
Step
Action
1
Switch on the instrument by pressing the Power switch to the I position.
Result: The instrument starts and the Instrument display states Not connected.
2
36
Turn on the computer and monitor according to the manufacturer's instructions.
Getting Started with ÄKTA avant 29-0351-83 AA
3 Start up
Start UNICORN and log on
Follow the instruction below to start UNICORN and log on to the program.
Step
Action
1
Double-click the UNICORN icon on the desktop.
Result: The Log On dialog opens.
Note:
If there is no connection to the database it is still possible to log on to UNICORN
and control a running system. The Log On dialog will give the option to start
System Control without a database. Click Start System Control to proceed
to the next Log On dialog.
Getting Started with ÄKTA avant 29-0351-83 AA
37
3 Start up
Step
Action
2
In the Log On dialog:
•
select User Name.
and
•
enter Password.
Note:
It is also possible to select the Use Windows Authentication checkbox
and enter a network ID in the User Name field.
•
click the Options button, and select which UNICORN modules to start.
•
click OK.
Result: The selected UNICORN modules open.
Connect to system
Follow the instructions below to connect the instrument to UNICORN.
38
Getting Started with ÄKTA avant 29-0351-83 AA
3 Start up
Step
Action
1
In the System Control module, click the Connect to Systems icon.
Result: The Connect to Systems dialog opens.
2
In the Connect to Systems dialog:
•
Select a system.
•
Select Control mode.
•
Click OK.
Result: The selected instrument can now be controlled by the software.
Getting Started with ÄKTA avant 29-0351-83 AA
39
4 The instrument and software
4
The instrument and software
About this chapter
This chapter gives an overview of ÄKTA avant: instrument, software and accessories.
Illustration of the system
The illustration below shows the ÄKTA avant instrument with UNICORN software installed
on a computer.
In this chapter
This chapter contains the following sections:
Section
40
See page
4.1 ÄKTA avant instrument overview
41
4.2 UNICORN software overview
44
4.3 UNICORN software modules
47
Getting Started with ÄKTA avant 29-0351-83 AA
4 The instrument and software
4.1 ÄKTA avant instrument overview
4.1
ÄKTA avant instrument overview
Introduction
This section shows an overview of the ÄKTA avant instrument. Technical details about
the instrument and the individual modules are found in the ÄKTA avant User Manual.
Exterior design
ÄKTA avant has a modular design, with all the liquid handling modules placed on the
exterior of the instrument. Buffer vessels are placed on the Buffer tray on top of the 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 operational limits of the instruments.
ÄKTA avant 25
Parameter
Limits
Flow rate
Up to 25 ml/min
Note! When running the Column packing
flow instruction, the maximum flow rate
for ÄKTA avant 25 is 50 ml/min.
Max. operating pressure
20 MPa
Wavelength
190 - 700 nm
ÄKTA avant 150
Parameter
Limits
Flow rate
Up to 150 ml/min
Note! When running the Column packing
flow instruction, the maximum flow rate
for ÄKTA avant 150 is 300 ml/min.
Max. operating pressure
Getting Started with ÄKTA avant 29-0351-83 AA
5 MPa
41
4 The instrument and software
4.1 ÄKTA avant instrument overview
Parameter
Limits
Wavelength
190-700 nm
Illustration of the main parts of
the instrument
The illustration below shows the location of the main parts of the instrument.
42
Part
Function
Part
Function
1
Fraction collector
7
Holder rails
2
Buffer tray
8
Swivel foot Lock/Unlock knob
3
Instrument display
9
Swivel foot
4
Wet side
10
Swing out toolbox
5
Foldable door
11
Power switch
6
Pump cover
Getting Started with ÄKTA avant 29-0351-83 AA
4 The instrument and software
4.1 ÄKTA avant 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
Function
Part
Function
1
Injection valve
11
Inlet valve A
2
Flow restrictor
12
System pump A
3
pH valve
13
Mixer
4
Column valve
14
Pressure monitor of Sample
pump
5
UV monitor
15
Sample pump
6
Conductivity monitor
16
Pump piston rinsing system
tubes
7
Inlet valve B
17
Sample inlet valve
8
Quaternary valve
18
Outlet valve
9
System pump B
19
Holder rails
10
Pressure monitor of System
pumps
Getting Started with ÄKTA avant 29-0351-83 AA
43
4 The instrument and software
4.2 UNICORN software overview
4.2
UNICORN software overview
Introduction
This section gives a brief overview of the UNICORN software: a complete package for
control, supervision and evaluation of chromatography instruments and purification
runs. It also describes how to access the help utility that is included in UNICORN.
UNICORN modules overview
UNICORN consists of four modules: Administration, Method Editor, System Control and
Evaluation. The main functions of each module are described in the table below.
Module
Main functions
Administration
Perform user and system setup, system log and database
administration.
Method Editor
Create and edit methods.
System Control
Start, view and control runs.
Evaluation
Open results, evaluate runs and create reports.
Enter a UNICORN module
To enter a module:
•
click the Taskbar button of the module of interest,
or
•
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.
44
Getting Started with ÄKTA avant 29-0351-83 AA
4 The instrument and 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...
find information about
a UNICORN module
select Help:Help for... in the UNICORN module of interest
find information about
the item currently selected and in focus (e.g., a
pane, a dialog, or a
method phase)
•
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
navigate the online help
search for a specific
term in the online help
Getting Started with ÄKTA avant 29-0351-83 AA
•
click the Help icon in the open dialog
•
select Help:Help for... in any of the UNICORN modules
(see illustration above)
•
in the TOC (Table of contents) pane, expand the
headings of interest to navigate the content structure
•
click the heading of interest to open a section
•
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
45
4 The instrument and software
4.2 UNICORN software overview
If you want to...
then...
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 6
online documentation portal and select 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:
•
open a method
•
select the instruction of interest in the Instruction box
in the Text instruction pane
•
press the F1 key
In the System Control module:
•
select Manual:Execute Manual Instructions
•
expand a heading and select the instruction of interest
•
press the F1 key
or
click the Help icon in the dialog
46
Getting Started with ÄKTA avant 29-0351-83 AA
4 The instrument and software
4.3 UNICORN software modules
4.3
UNICORN software modules
About this section
Three of the four UNICORN modules are used in this Getting Started guide and described
in this section: Method Editor, System Control and Evaluation. The Administration
module and its icons are described in UNICORN 6 Administration and Technical Manual.
In this section
This section contains the following subsections:
Section
See page
4.3.1 Method Editor module
48
4.3.2 System Control module
51
4.3.3 Evaluation module
53
Getting Started with ÄKTA avant 29-0351-83 AA
47
4 The instrument and software
4.3 UNICORN software modules
4.3.1 Method Editor module
4.3.1
Method Editor module
Introduction
In UNICORN software, the instructions to control a chromatography run are defined in
a method. The Method Editor module is used to create or edit such methods. The
graphical user interface combines default settings with the ability to change settings.
This simplifies method creation and replaces the method wizard in earlier versions of
UNICORN.
Predefined methods and phases
A method is built up by a number of phases. Each phase represents a major process
step in the method, for example, equilibration or elution. Predefined methods, that include
all the phases necessary to run the system, are available for different chromatography
techniques and also for system cleaning. Refer to UNICORN 6 Method Manual for information about predefined methods and phases and how to create a method.
48
Getting Started with ÄKTA avant 29-0351-83 AA
4 The instrument and software
4.3 UNICORN software modules
4.3.1 Method Editor module
Method Editor panes
As illustrated below, four panes show in the Method Editor by default. Available phases
are found in the Phase Library (1), and an overview of the phases included in the active
method is displayed in a Method Outline (2). Detailed information about the method is
presented in the upper right-hand pane (3), containing the two tabs Phase Properties
and Text Instructions. The gradient used in the method is displayed in a Gradient illustration (4).
1
2
3
4
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.
Getting Started with ÄKTA avant 29-0351-83 AA
Icon
Function
Open Method Navigator:
Opens the Method Navigator where available methods are listed.
49
4 The instrument and software
4.3 UNICORN software modules
4.3.1 Method Editor module
Icon
50
Function
Icon
Function
Save: Saves the active
method.
Print: Opens the Print dialog from where a method
can be printed.
Copy: Copies the selected
method or folder.
Paste: Pastes a copied
method or folder to a new
location.
Undo: Restores the method
to the state it was in before
the last change.
Redo: Restores the method
to the state it was in before
the Undo command was
used.
Getting Started with ÄKTA avant 29-0351-83 AA
4 The instrument and software
4.3 UNICORN software modules
4.3.2 System Control module
4.3.2
System Control module
Introduction
The System Control module is used to start, view, and control a method run.
System Control panes
As illustrated below, four panes show in the System Control by default. The Run Data
pane (1) presents current data in numerical values, while the Chromatogram pane (2)
illustrates data as curves during the entire method run. Information about the method
progression is presented in the Run Log (3) and the current flow path is illustrated in the
Flow Scheme (4).
1
2
3
Getting Started with ÄKTA avant 29-0351-83 AA
4
51
4 The instrument and software
4.3 UNICORN software modules
4.3.2 System Control module
System Control toolbar icons
The table below shows the System Control toolbar icons that are referred to in this
Getting Started.
Icon
52
Function
Icon
Function
Open Method Navigator:
Opens the Method Navigator where available methods are listed.
Run: Starts a method run.
Hold: Suspends the method
run, while current flow rate
and valve positions are
sustained.
Pause: Suspends the
method run and stops all
pumps.
Continue: Resumes for example a held or paused
method run.
End: Permanently ends the
method run.
Customize: Opens the Customize dialog where curve
settings, run data groups
and run log contents can be
set.
Connect to Systems: Opens
the Connect to Systems dialog where systems can be
connected, and currently
connected users are displayed.
Getting Started with ÄKTA avant 29-0351-83 AA
4 The instrument and software
4.3 UNICORN software modules
4.3.3 Evaluation module
4.3.3
Evaluation module
Introduction
The Evaluation module is used to evaluate the results from 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.
1
2
3
Getting Started with ÄKTA avant 29-0351-83 AA
53
4 The instrument and software
4.3 UNICORN software modules
4.3.3 Evaluation module
Evaluation toolbar icons
The table below shows the Evaluation toolbar icons that are referred to in this Getting
Started.
Icon
54
Function
Icon
Function
Open Result Navigator:
Opens the Result Navigator
where available results are
listed.
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.
Undo: Restores the result 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.
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.
Getting Started with ÄKTA avant 29-0351-83 AA
5 Create a method
5
Create a method
About this chapter
A method is created in the Method Editor module. This chapter describes method creation
based on predefined methods, and also how to print a method. Please refer to UNICORN
6 Method Manual for more detail about method creation.
In this chapter
This chapter contains the following sections:
Section
See page
5.1 Guide to method creation
56
5.2 Print a method
68
Getting Started with ÄKTA avant 29-0351-83 AA
55
5 Create a method
5.1 Guide to method creation
5.1
Guide to method creation
About this section
This section describes how to create a new method based on a predefined method, and
how to edit a method. The section also provides an overview of the predefined methods
and phases that are available.
For further information about method creation, see UNICORN 6 Method Manual.
In this section
This section contains the following subsections:
Section
56
See page
5.1.1 Create and edit methods
57
5.1.2 Predefined methods
65
5.1.3 Predefined phases
67
Getting Started with ÄKTA avant 29-0351-83 AA
5 Create a method
5.1 Guide to method creation
5.1.1 Create and edit methods
5.1.1
Create and edit methods
Introduction
Method creation in UNICORN 6 is performed in the Method Editor module.
Predefined methods are available for a number of different chromatography techniques,
for example affinity chromatography and gel filtration. There are also predefined methods
for column preparation and system cleaning.
A method is comprised of a number of phases. Each phase represents a major process
step in the method, for example, equilibration or elution. If desired, a predefined method
can be edited by adding, deleting and/or rearranging phases, and also by editing the
settings of each phase included in the method.
Illustration of the Method Editor
user interface
The illustration and table below show the Method Editor module and its panes.
1
2
3
4
Area
Description
1
Phase Library: Displays all available phases.
Getting Started with ÄKTA avant 29-0351-83 AA
57
5 Create a method
5.1 Guide to method creation
5.1.1 Create and edit methods
Area
Description
2
Method Outline: Presents an overview of the phases included in the active
method.
3
Phase Properties: Presents detailed settings for the selected phase.
4
Gradient: Illustrates the gradient used in the active method.
Create a method
Follow the instruction below to create and edit a chromatographic method based on a
predefined method.
Step
Action
1
Open the Method Editor module and click the New Method icon.
Result: The New Method dialog opens.
2
Select System and Predefined Method in the dialog. Click OK.
Result: The phases included in the chosen method is shown in the Method
Outline pane, and the default settings for each of the phases is shown in
the Phase Properties pane.
58
Getting Started with ÄKTA avant 29-0351-83 AA
5 Create a method
5.1 Guide to method creation
5.1.1 Create and edit methods
Step
Action
3
In the Phase Properties pane of the Method Settings phase, edit general
settings like Column type and Method Base Unit. UNICORN automatically
calculates correct settings for volume, flow rate, and pressure limits based
on the selected column type.
The illustration below shows the Method Outline pane and the Phase
Properties pane of the Method Settings phase.
Getting Started with ÄKTA avant 29-0351-83 AA
59
5 Create a method
5.1 Guide to method creation
5.1.1 Create and edit methods
Step
Action
4
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.
Result: The Result Name & Location dialog opens.
In the Result Name & Location dialog:
60
•
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.
•
Select Result name. With the default selection Method name the results
will be named with the method name together with a serial number.
•
Click OK to confirm and close the dialog.
Getting Started with ÄKTA avant 29-0351-83 AA
5 Create a method
5.1 Guide to method creation
5.1.1 Create and edit methods
Step
Action
5
In the Phase Properties pane of the Method Settings phase, click the Start
Protocol button if you wish to include a start protocol.
Result: The Start Protocol dialog opens.
In the Start Protocol dialog:
•
Select items to display at method start. Result Name and Location is
selected by default.
•
Click OK to confirm and close the dialog.
6
Select the next phase in the Method Outline pane by clicking it, and choose
appropriate parameter values in the Phase Properties pane.
7
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 ÄKTA avant 29-0351-83 AA
61
5 Create a method
5.1 Guide to method creation
5.1.1 Create and edit methods
Step
Action
8
Click the Save the method icon.
Result: The Save As dialog opens.
9
In the Save As dialog:
•
Select a target folder to enable the Save button.
•
Type a Name for the method.
•
Select a System from the list.
•
Click the Save button.
Result: The created method is saved in the selected folder.
Add phases to a method
Follow the instruction below to add phases to a method.
Step
Action
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.
62
Getting Started with ÄKTA avant 29-0351-83 AA
5 Create a method
5.1 Guide to method creation
5.1.1 Create and edit methods
Delete phases from a method
Follow the instruction below to delete phases from a method.
Step
Action
1
Select the phase to delete in the Method Outline pane.
Note:
The Method Settings phase cannot be deleted.
2
Click the Delete button located below the Method Outline pane.
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.
Choose a maintenance method
A number of predefined methods for preparation and cleaning are available. Use these
maintenance methods to prepare and clean the system and columns, and to fill the
system and columns with storage solution. The table below gives suggestions for what
methods and solutions to choose for the different purposes.
If you wish to...
Phase/Method
Solution
Prepare tubing and instrument modules
before a run
System Preparation
suitable buffers
Prepare column before a run
Column Preparation
suitable buffers
Getting Started with ÄKTA avant 29-0351-83 AA
63
5 Create a method
5.1 Guide to method creation
5.1.1 Create and edit methods
64
If you wish to...
Phase/Method
Solution
Clean tubing and modules
System CIP
0.5-1 M NaOH
Clean column
Column CIP
0.5-1 M NaOH
Leave system in storage solution
System CIP
20% Ethanol
Leave column in storage solution
Column CIP
20% Ethanol
Tip:
CIP (Cleaning-In-Place) and preparation of columns and system may be run
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 ÄKTA avant 29-0351-83 AA
5 Create a method
5.1 Guide to method creation
5.1.2 Predefined methods
5.1.2
Predefined methods
The predefined methods that are available are described in the table below.
Method
Description
Affinity 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 reequilibrated with start 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
Chromatography
(CIEX)
After equilibration and sample application, positively 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.
Chromatofocusing (CF)
After equilibration and sample application, elution is performed
using a pH gradient. The proteins separate and elute according
to their isoelectric points. Finally, the column is re-equilibrated.
Only available for ÄKTA avant 25.
Column CIP
The column is filled with a cleaning solution. Select inlet positions.
Enter the solution identity, volume, flow rate and incubation time.
By adding steps, several cleaning solutions can be used. Suggestions for cleaning steps are available for a number of column
types.
Column Performance Test
After equilibration of the column, sample is injected and eluted
isocratically. A non-adsorbing sample like acetone or salt should
be used. After the run, calculate column performance in the
Evaluation module. The efficiency of the column is determined
in terms of height equivalent to a theoretical plate (HETP), and
the peak asymmetry factor (As). 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.
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65
5 Create a method
5.1 Guide to method creation
5.1.2 Predefined methods
Method
Description
Desalting (DS)
After equilibration and sample application, the proteins are
eluted isocratically. This technique is commonly used for buffer
exchange.
Gel Filtration (GF)
After equilibration and sample application, proteins separate
and elute according to their size (largest first).
Hydrophobic 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.
Intelligent Packing
Packs AxiChrom™ columns, with a predetermined column type,
by a flow of hydraulic liquid that pushes the adaptor down. The
user initiates the start of compression at the exact point when
the adapter reaches the consolidated bed surface. The adapter
compresses the bed according to the packing factor or target
bed height as selected. Two Column Performance Test (upflow/downflow) phases are automatically performed after the
AxiChrom column has been packed.
Only available for ÄKTA avant 150.
66
Reversed Phase
Chromatography
(RPC)
After equilibration and sample application, hydrophobic proteins
adsorb to the column ligand. After a wash to remove unbound
sample, elution is performed by generating a gradient of a nonpolar, 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 ÄKTA avant 29-0351-83 AA
5 Create a method
5.1 Guide to method creation
5.1.3 Predefined phases
5.1.3
Predefined phases
The predefined phases that are available in the predefined methods and in the Phase
Library are described in the table below.
Phase
Description
Method Settings
The first, and mandatory, phase in any method. Defines 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 handling of flowthrough.
Column Wash
Washes out unbound sample after sample application or removes strongly bound proteins after elution.
Elution
Elutes the sample from the column. Defines parameters for
the elution and fractionation settings.
Column Preparation
Prepares the column before use by removing the storage 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 Preparation
Prepares the system before a run by removing storage solution
and filling the system and inlets with buffer solution. One
preparation solution is used per phase.
System CIP
Cleans the system after purification runs by rinsing the system
with a cleaning solution. One cleaning solution is used per
phase.
Column Performance Test
Tests the efficiency of a packed column in terms of height
equivalent to a theoretical plate (HETP), and the peak asymmetry factor (As).
Intelligent Packing
A flow of hydraulic liquid pushes the adapter down. The user
initiates the start of compression at the exact point when the
adapter reaches the consolidated bed surface. The adapter
compresses the bed according to the packing factor or target
bed height as selected.
Only available for ÄKTA avant 150.
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67
5 Create a method
5.2 Print a method
5.2
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.
Step
Action
1
Open the Method Editor module and click the Open Method Navigator icon
in the toolbar.
Result: The Method Navigator pane opens.
2
Select the method to print and click the Open a Method icon in the navigator
toolbar.
Result: The selected method opens.
68
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5 Create a method
5.2 Print a method
Step
Action
3
Click the Print icon.
Result: The Print dialog opens.
4
In the Print dialog:
•
select Printer
•
click OK
Result: The method is printed.
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69
6 Prepare the system for a run
6
Prepare the system for a run
About this chapter
This chapter describes the preparations necessary to prepare the system before starting
a run.
WARNING
Always use appropriate personal protective equipment during operation and maintenance of ÄKTA avant system.
In this chapter
This chapter contains the following sections:
Section
6.1 Before you prepare the system
71
6.2 Prepare the flow path
73
6.3 Prime buffer inlets and purge system pumps
81
6.4 Connect a column
89
6.5 Calibrate the pH monitor
94
6.6 Prepare the Fraction collector
96
6.7 Prepare for a run in a cold cabinet
70
See page
102
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6 Prepare the system for a run
6.1 Before you prepare the system
6.1
Before you prepare the system
Introduction
It is important to prepare the system in accordance with the settings in the method to
be run. Before preparing the system, check the settings in the Method Editor and make
sure that all accessories to be used are available.
Checklist
Make sure to prepare the system in accordance with the settings in the method to be
run. Remember to check:
•
which valve ports to use for inlets and outlets
•
which column type to use
•
which column position to use
•
which buffers and samples to prepare
•
which sample application technique to use
•
that the pH electrode is connected, if applicable
•
which cassettes with corresponding deep well plates and/or tubes to use in the
Fraction collector, if applicable
•
if it is a reversed phase chromatography (RPC) run
WARNING
Fraction collector. Do not fractionate flammable liquids. When
running RPC methods, or other procedures using organic solvents, collect fractions through the Outlet valve.
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6 Prepare the system for a run
6.1 Before you prepare the system
WARNING
RPC runs with 100% acetonitrile in ÄKTA avant 25. Always
replace the green PEEK tubing between the used system pump
and the pump pressure monitor with orange PEEK tubing, i.d.
0.5 mm, before running RPC with 100% acetonitrile. Set the
System pressure alarm to 10 MPa.
RPC runs with 100% acetonitrile in ÄKTA avant 150. Always
replace the beige PEEK tubing between the used system pump
and the pump pressure monitor with orange PEEK tubing, i.d.
0.5 mm, before running RPC with 100% acetonitrile.
72
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6 Prepare the system for a run
6.2 Prepare the flow path
6.2
Prepare the flow path
Introduction
The flow path contains tubing, valves, pumps and monitors. This section gives an overview
of the flow path and describes how to prepare the flow 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.
9
10
11
8
15
12
14
13
16
5
1
3
2
Part
Description
1
Sample inlet valve
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6
4
7
73
6 Prepare the system for a run
6.2 Prepare the flow path
Part
Description
2
Sample pump
3
Inlet valve A
4
System pump A
5
Quaternary valve
6
Inlet valve B
7
System pump B
8
Injection valve
9
Capillary loop or Superloop
10
Column valve
11
Column
12
UV monitor
13
Conductivity monitor
14
pH valve with pH monitor and Flow restrictor
15
Outlet valve
16
Fraction collector
Tubing and connectors
The table below shows what types of tubing and connectors to use for different purposes.
ÄKTA avant 25
74
Scope of use
Tubing
Connector
Photo
Inlet tubing
FEP, o.d. 1/8",
i.d. 1.6 mm
Tubing connector,
5/16" + Ferrule (yellow),
1/8"
Tubing to connect columns
PEEK, o.d.
1/16", i.d. 0.50
mm
Fingertight connector,
1/16"
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6 Prepare the system for a run
6.2 Prepare the flow path
Scope of use
Tubing
Connector
Outlet and
waste tubing
ETFE, o.d.
1/16", i.d. 1.0
mm
Fingertight connector,
1/16"
Scope of use
Tubing
Connector
Inlet tubing
FEP, o.d. 3/16",
i.d. 2.9 mm
Tubing connector,
5/16" + Ferrule (blue),
3/16"
Tubing to connect columns
PEEK, o.d.
1/16", i.d. 1.0
mm
Fingertight connector,
1/16"
Outlet tubing
FEP, o.d. 1/8",
i.d. 1.6 mm
Tubing connector,
5/16" + Ferrule (yellow),
1/8"
Waste tubing
ETFE, o.d.
1/16", i.d. 1.0
mm
Fingertight connector,
1/16"
Photo
ÄKTA avant 150
Getting Started with ÄKTA avant 29-0351-83 AA
Photo
75
6 Prepare the system for a run
6.2 Prepare the flow path
Inlet ports
The table below shows the inlet ports of Inlet valve A, Inlet valve B and Sample inlet valve.
Valves and ports
Illustrations
Inlet valve A
Inlet ports:
•
A1-A7
Inlet valve B
Inlet ports:
•
B1-B7
Sample inlet valve
Inlet ports:
•
76
S1-S7, Buff
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6 Prepare the system for a run
6.2 Prepare the flow path
Prepare the inlet tubing
Connect inlet tubing to the inlet ports that are to be used, and immerse all inlet tubing
that is to be used during the method run in the correct buffers.
Prepare the outlet tubing
Connect outlet tubing to the outlet ports of Outlet valve that are to be used during the
run. If a fraction collector is to be used, make sure that tubing is connected between the
Outlet valve Frac port and the Fraction collector, and prepare the fraction collector.
Otherwise, immerse the outlet tubing in suitable tubes or flasks.
Waste ports
The table below shows the waste ports of Injection valve, pH valve, and Outlet valve.
Valves and ports
Illustrations
Injection valve
Waste ports:
•
W1, W2
pH valve
Waste port:
•
W3
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6 Prepare the system for a run
6.2 Prepare the flow path
Valves and ports
Illustrations
Outlet valve
Waste port:
•
W
Location of waste tubing
All waste tubing is found on the rear of the instrument, see the illustration below.
1
78
2
Part
Description
1
Waste tubing from the Injection valve, the pH valve and the Outlet valve
(W, W1, W2 and W3).
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6 Prepare the system for a run
6.2 Prepare the flow path
Part
Description
2
Waste tubing from the Fraction collector and the Buffer tray.
Prepare the waste tubing
Follow the instruction below to prepare the waste tubing.
Step
Action
1
Make sure that the four pieces of waste tubing from the Injection valve, the
pH valve and the Outlet valve (W, W1, W2 and W3) are placed in a vessel
placed below the bench.
NOTICE
The maximum level of the waste vessel for the waste
tubing from the valves must be lower than 30 cm
above the lab bench.
2
Make sure that the three pieces of waste tubing from the Fraction collector
and the Buffer tray are placed in a waste vessel placed below the bench.
NOTICE
The maximum level of the waste vessel for the waste
tubing from the Fraction collector and the Buffer tray
must be lower than the bench height.
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6 Prepare the system for a run
6.2 Prepare the flow path
Step
Action
3
Make sure that the waste tubing from the Fraction collector and the Buffer
tray is cut to appropriate length. It is important that the tubing is not bent
and will not be submerged in liquid during the run.
Note:
If the tubing is too short, replace it with new tubing. Do not lengthen the tubing
as this might cause obstruction of the tubing and flooding in the Frac chamber.
CAUTION
Make sure that the waste vessels will hold all the produced volume
of the run. For ÄKTA avant 25, a suitable waste vessel should typically have a volume of 2 to 10 liters. For ÄKTA avant 150, a waste
vessel should have a volume of 40 liters.
80
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6 Prepare the system for a run
6.3 Prime buffer inlets and purge system pumps
6.3
Prime buffer inlets and purge system pumps
Introduction
Before usage of the System pumps, it is important to:
1
Prime the inlets (fill the buffer inlets with liquid).
2
Purge the System pumps (remove air from the pump heads).
Overview
The procedure consists of the following stages:
1
Prime all inlet tubing B to be used during the run
2
Prime all inlet tubing A to be used during the run
3
Prepare the system before purging the System pumps
4
Purge System pump B
5
Purge System pump A
6
End the run
Prime inlet tubing B
Follow the instructions below to fill all B inlet tubing to be used in the run with appropriate
buffer/solution.
Step
Action
1
Make sure that all B inlet tubing that is to be used during the method run is
immersed in the correct buffers.
Getting Started with ÄKTA avant 29-0351-83 AA
81
6 Prepare the system for a run
6.3 Prime buffer inlets and purge system pumps
Step
Action
2
In the Manual instructions dialog:
•
Select Flow path:Inlet B.
•
Select the Position of the inlet to be filled from the drop-down list. Start
at the inlet position with the highest number and end at the inlet position
with the lowest number.
•
Click
.
Result: Inlet valve B switches to the selected port.
82
3
Connect a 25-30 ml syringe to the purge valve of one of the pump heads of
System pump B. Make sure that the syringe fits tightly into the purge connector.
4
Open the purge valve by turning it counter-clockwise about 3 quarters of a
turn. Draw liquid slowly into the syringe until the liquid reaches the pump.
Getting Started with ÄKTA avant 29-0351-83 AA
6 Prepare the system for a run
6.3 Prime buffer inlets and purge system pumps
Step
Action
5
Close the purge valve by turning it clockwise. Disconnect the syringe and
discard its contents.
6
Repeat steps 2-5 for each piece of inlet tubing B that is to be used during
the run.
Prime inlet tubing A
Follow the instruction below to fill all A inlet tubing, to be used in the run, with appropriate
buffer/solution.
Step
Action
1
Make sure that all A inlet tubing that is to be used during the method run is
immersed in the correct buffers.
2
In the Manual instructions dialog:
•
Select Flow path:Inlet A.
•
Select the Position of the inlet to be filled from the drop-down list.
•
Click
.
Result: Inlet valve A switches to the selected port.
3
Connect a 25-30 ml syringe to the purge valve of one of the pump heads of
System pump A. Make sure that the syringe fits tightly into the purge connector.
4
Open the purge valve by turning it counter-clockwise about 3 quarters of a
turn. Draw liquid slowly into the syringe until the liquid reaches the pump.
5
Close the purge valve by turning it clockwise. Disconnect the syringe and
discard its contents.
6
Repeat steps 2-5 for each piece of inlet tubing A that is to be used during
the run.
Getting Started with ÄKTA avant 29-0351-83 AA
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6 Prepare the system for a run
6.3 Prime buffer inlets and purge system pumps
Prepare the system before
purging the System pump heads
Follow the instructions below to prepare the system.
Step
Action
1
Make sure that the piece of waste tubing connected to Injection valve port
W1 is placed in a waste vessel.
2
Open the System Control module and select Manual:Execute Manual Instructions.
Result: The Manual instructions dialog opens.
3
In the Manual instructions dialog:
•
Select Flow path:Injection valve and select System pump waste from
the Position drop-down list. Click
.
Result: The Injection valve switches to waste position. This is necessary
to achieve a low backpressure during the purge procedure.
•
Select Pumps and pressures:System flow. For ÄKTA avant 25, set the
Flow rate to 1.0 ml/min. For ÄKTA avant 150, set the Flow rate to 10.0
ml/min. Click
.
Result: A system flow starts.
84
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6 Prepare the system for a run
6.3 Prime buffer inlets and purge system pumps
Purge System pump B
Follow the instruction below to purge both pump heads of System pump B.
Step
Action
1
In the Manual instructions dialog:
•
Select Pumps and pressures:Gradient.
•
Set Target to 100% B and Length to 0 min.
•
Click
.
Result: Only System pump B is active.
2
In the Manual instructions dialog:
•
Select Flow path:Inlet B.
•
Select the Position of one of the inlets to be used from the drop-down
list.
•
Click
.
Result: Inlet valve B switches to the selected port.
Getting Started with ÄKTA avant 29-0351-83 AA
85
6 Prepare the system for a run
6.3 Prime buffer inlets and purge system pumps
86
Step
Action
3
Connect a 25 to 30 ml syringe to the purge valve of the left pump head of
System pump B. Make sure that the syringe fits tightly into the purge connector.
4
Open the purge valve by turning it counter-clockwise about 3 quarters of a
turn. Draw a small volume of liquid slowly into the syringe (with a rate of
about 1 ml per second).
5
Close the purge valve by turning it clockwise. Disconnect the syringe and
discard its contents.
6
Connect the syringe to the purge valve on the right pump head of System
pump B, and repeat steps 4 and 5. Keep the system flow running.
Getting Started with ÄKTA avant 29-0351-83 AA
6 Prepare the system for a run
6.3 Prime buffer inlets and purge system pumps
Purge System pump A
Follow the instructions below to purge both pump heads of System pump A.
Step
Action
1
In the Manual instructions dialog:
•
Select Pumps and pressures:Gradient.
•
Set Target to 0% B and Length to 0 min.
•
Click
.
Result: Only System pump A is active.
2
In the Manual instructions dialog:
•
Select Flow path:Inlet A.
•
Select the Position of one of the inlets to be used from the drop-down
list.
•
Click
.
Result: Inlet valve A switches to the selected port.
3
Connect a 25-30 ml syringe to the purge valve of the left pump head of
System pump A. Make sure that the syringe fits tightly into the purge connector.
4
Open the purge valve by turning it counter-clockwise about 3 quarters of a
turn. Draw a small volume of liquid slowly into the syringe (with a rate of
about 1 ml per second).
Getting Started with ÄKTA avant 29-0351-83 AA
87
6 Prepare the system for a run
6.3 Prime buffer inlets and purge system pumps
Step
Action
5
Close the purge valve by turning it clockwise. Disconnect the syringe and
discard its contents.
6
Connect the syringe to the purge valve on the right pump head of System
pump A, and repeat steps 5-6. Keep the system flow running.
End the run
Click the End icon in the System Control toolbar to end the run.
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6 Prepare the system for a run
6.4 Connect a column
6.4
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 ÄKTA avant.
WARNING
To avoid exposing the column to excessive pressure, make sure
that the pressure limit is set to the specified maximum pressure of
the column. Before connecting a column to the ÄKTA avant instrument, read the instructions for use of the column.
Methods automatically include a pressure alarm based on the specifications of the
chosen column type. However, when running manual runs you have to set the pressure
limits yourself.
Note:
Do not overtighten when connecting columns. Overtightening might rupture
the connectors or squeeze the tubing and thereby result in high back pressure.
Attach a column holder and
connect a column
Follow the instruction below to connect a column to the instrument. Always use a column
holder. The column is connected to two opposite parts of the Column valve, using appropriate tubing and connectors.
Step
Action
1
Choose a column according to the column selections made in the method
to be run.
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89
6 Prepare the system for a run
6.4 Connect a column
Step
Action
2
Attach an appropriate column holder to the rail on the instrument.
3
Attach the column to the column holder.
4
Connect a suitable tubing to a Column valve port, for example port 2A if
column position 2 was chosen in the method to be run.
5
Open the System Control module and select Manual:Execute Manual Instructions.
Result: The Manual instructions dialog opens.
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6 Prepare the system for a run
6.4 Connect a column
Step
Action
6
In the Manual instructions dialog:
•
Select Flow path:Column position.
•
Choose the Position used for connection of the column, in this example
Position 2.
•
Choose the Flow Direction Down flow.
•
Click
.
Result: The Column valve switches to position 2.
7
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.
Getting Started with ÄKTA avant 29-0351-83 AA
91
6 Prepare the system for a run
6.4 Connect a column
92
Step
Action
8
When buffer leaves the tubing on port 2A in a continuous mode and the top
part of the column is filled with buffer, connect the tubing to the top of the
column.
9
Connect a piece of tubing to the bottom of the column.
10
When buffer leaves the tubing at the bottom of the column in a continuous
mode, connect this piece of tubing to the Column valve. Use the port opposite
to the one already connected to the column, in this example port 2B.
Getting Started with ÄKTA avant 29-0351-83 AA
6 Prepare the system for a run
6.4 Connect a column
Step
Action
11
Click the End icon in the System Control toolbar to end the run.
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93
6 Prepare the system for a run
6.5 Calibrate the pH monitor
6.5
Calibrate the pH monitor
Introduction
If pH will be measured during the chromatographic run, the pH monitor should be 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
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.
2
3
Set the pH monitor as the Monitor to calibrate by selecting pH from the list.
Click
.
Result: The pH valve switches to the calibration position.
94
4
Enter the pH of the first pH standard buffer in the pH for buffer 1 field.
5
Fill a syringe with approximately 10 ml of the first pH standard buffer. Connect the syringe to the Luer connector in pH valve port Cal, and inject the
buffer.
Getting Started with ÄKTA avant 29-0351-83 AA
6 Prepare the system for a run
6.5 Calibrate the pH monitor
Step
6
Action
When the Current value is stable, click
.
7
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 ≥ 80% and the Asymmetry potential at
pH 7 inside the interval ±60 mV?
•
•
If Yes: Click
to switch the pH valve back to the default position,
and to close the Calibration dialog.
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.
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95
6 Prepare the system for a run
6.6 Prepare the Fraction collector
6.6
Prepare the Fraction collector
Introduction
This section describes how to prepare the Fraction collector. For information regarding
the types of deep well plates, tubes and Cassettes, see ÄKTA avant User Manual.
WARNING
Fraction collector. Do not fractionate flammable liquids. When
running RPC methods, or other procedures using organic solvents,
collect fractions through the Outlet valve.
Prepare the Fraction collector
Follow the instructions below to prepare the Fraction collector before a run.
Cassettes and Cassette tray
96
Step
Action
1
If you are to use Cassettes with the QuickRelease function, open the Cassettes.
Getting Started with ÄKTA avant 29-0351-83 AA
6 Prepare the system for a run
6.6 Prepare the Fraction collector
Step
Action
2
Place the tubes and deep well plates in the Cassettes. Make sure that the
deep well plates are rotated so that the well marked A1 is positioned above
the A1 marking on the Cassette.
3
Close the Cassettes that have the QuickRelease function.
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6 Prepare the system for a run
6.6 Prepare the Fraction collector
98
Step
Action
4
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.
5
Open the Frac drawer by pressing the handle upwards, and pulling out the
drawer.
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.
Getting Started with ÄKTA avant 29-0351-83 AA
6 Prepare the system for a run
6.6 Prepare the Fraction collector
Step
Action
7
Close the Frac drawer. Make sure that it snaps into closed position.
50 ml tube tray
Step
Action
1
Place 50 ml tubes in the 50 ml tube tray.
2
Open the Frac drawer by pressing the handle upwards, and pulling out the
drawer.
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6 Prepare the system for a run
6.6 Prepare the Fraction collector
Step
Action
3
Place the 50 ml tube tray on the Tray support of the Frac drawer. Make sure
that the front of the tray (marked with the GE logo) faces the front of the
drawer and is hooked onto the two pins.
Note:
The Cassette tray shall not be used when the 50 ml tube tray or the 250 ml
bottle tray is placed in the Frac drawer.
4
Close the Frac drawer. Make sure that it snaps into closed position.
Cassette identification
After the Frac drawer has been closed, the Frac arm scans the Cassette type code of
each Cassette to identify the Cassette types. If deep well plates are used, the instrument
also identifies the types of deep well plates.
WARNING
Moving parts in Fraction collector. Do not open the Fraction 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.
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6 Prepare the system for a run
6.6 Prepare the Fraction collector
View Fraction collector content
To view the content of the Fraction collector, open the System control module and select
View:Fraction Collector Content.
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6 Prepare the system for a run
6.7 Prepare for a run in a cold cabinet
6.7
Prepare for a run in a cold cabinet
Introduction
To fit the ÄKTA avant instrument in a cold cabinet, the foldable door and pump cover
can be removed. For instructions, see ÄKTA avant User Manual. When using the instrument
in a cold room or cold cabinet, make sure to follow the precautions listed below.
Precautions concerning runs in
a cold cabinet
NOTICE
Avoid condensation. If ÄKTA avant is kept in a cold room, cold
cabinet or similar, keep it switched on in order to avoid condensation.
NOTICE
Avoid overheating. If ÄKTA avant is kept in a cold cabinet and the
cold cabinet is switched off, make sure to switch off ÄKTA avant
and keep the cold cabinet open to avoid overheating.
NOTICE
Place the computer in room temperature. If the ÄKTA avant instrument is placed in a cold room, use a cold room compatible
computer or place the computer outside the cold room and use
the Ethernet cable delivered with the instrument to connect to the
computer.
Note:
102
When the instrument is kept in a cold room, it is important to tighten all tubing
connectors, also the inlet manifold connectors. Otherwise air might get into
the flow path.
Getting Started with ÄKTA avant 29-0351-83 AA
6 Prepare the system for a run
6.7 Prepare for a run in a cold cabinet
Tip:
When runs are performed in a cold cabinet, make sure to adjust the target
temperature of the Fraction collector temperature control function. The target
temperature is 20ºC by default. Settings for the temperature control function
can be edited in the System Settings dialog of System Control, or in the Text
Instructions pane in Method Editor.
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7 Run a method
7
Run a method
About this chapter
This chapter describes how to start up and run a method, and also how to handle the
system after the run.
In this chapter
This chapter contains the following sections:
Section
104
See page
7.1 Before you start
105
7.2 Applying the sample
108
7.3 Start a method run
111
7.4 Monitor the run
116
7.5 After run procedures
121
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7 Run a method
7.1 Before you start
7.1
Before you start
Introduction
Before starting a run, it is necessary to read and understand the information in this
section and to perform the checks listed below.
WARNING
Always use appropriate personal protective equipment during operation and maintenance of ÄKTA avant.
WARNING
Hazardous substances. When using hazardous chemical and biological agents, take all suitable protective measures, such as
wearing protective glasses and gloves resistant to the substances
used. Follow local and/or national regulations for safe operation,
maintenance and decommissioning of the equipment.
WARNING
High pressure. ÄKTA avant operates under high pressure. Wear
protective glasses at all times.
Checklist
Make sure that the system is correctly prepared. Check that:
•
The system is prepared according to the settings in the method to be run.
•
A suitable column has been selected for the application (consider target protein and
pressure range).
•
The buffer inlet tubing is immersed in correct buffer vessels (consider solution identity and volume).
•
All waste tubing is immersed in appropriate waste vessels (consider vessel size,
placement and material).
•
No tubing is twisted and the flow path is free from leakage.
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7 Run a method
7.1 Before you start
Hold, pause or stop the run
At the end of a method the run stops automatically. All pumps stop and an acoustic end
signal sounds and End is displayed in the Run Log.
To interrupt a method during a run you may use the Hold, Pause or End icons in System
Control. A held or paused method run can be resumed by using the Continue icon. See
the instructions in the table below.
If you want to...
then...
temporarily hold the method, with current
flow rate and valve positions sustained
click the Hold icon.
temporarily pause the method, and stop
all pumps
click the Pause icon.
resume, for example, a held or paused
method run.
click the Continue icon.
Note:
An ended method cannot be continued.
permanently end the run
Note:
106
click the End icon.
When ending a method run in advance, it is possible to save the partial result.
Getting Started with ÄKTA avant 29-0351-83 AA
7 Run a method
7.1 Before you start
Warnings concerning use of
hazardous substances
WARNING
Hazardous chemicals during run. When using hazardous chemicals, run System CIP and Column CIP to flush the entire system
tubing with distilled water, before service and maintenance.
WARNING
Hazardous biological agents during run. When using hazardous
biological agents, run System CIP and Column CIP to flush the
entire system tubing with bacteriostatic solution (e.g., NaOH) followed by a neutral buffer and finally distilled water, before service
and maintenance.
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7 Run a method
7.2 Applying the sample
7.2
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.
108
Stage
Description
Load
The capillary loop is filled with sample.
Inject
The sample is injected onto the column.
Getting Started with ÄKTA avant 29-0351-83 AA
7 Run a method
7.2 Applying the sample
Sample application through a
capillary loop
A capillary loop is manually filled with sample using a syringe connected to the Injection
valve port Syr. During the method run, the sample is automatically injected onto the
column. The loop is emptied and washed out using buffer from the system pumps. The
total buffer volume to be used for emptying and washing the capillary loop is set in the
Phase Properties tab of the Sample Application phase at Empty loop with.
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.
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7 Run a method
7.2 Applying the sample
How to fill a capillary loop
Follow the instruction below to fill the capillary loop with sample.
Step
Action
1
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.
4
Load sample into the capillary loop. To avoid sample loss due to siphoning,
leave the syringe in the port until the sample has been injected onto the
column during the run.
Tip:
It is recommended to overload the loop to make sure that the loop is completely filled. Excess of sample will leave the valve through port W1.
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7 Run a method
7.3 Start a method run
7.3
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.
Result: The Method Navigator pane opens.
2
Select the method to run, and click the Run icon.
Result: The Start Protocol dialog opens.
3
Step through the displayed pages in the Start Protocol, add requested input
and make appropriate changes if necessary. Click Next.
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7 Run a method
7.3 Start a method run
Step
Action
4
Click Finish on the last page of the Start Protocol.
Result:
5
6
•
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.
•
If column logging was not chosen at installation of UNICORN and/or no
column type was selected at method creation, the run starts directly.
Is the column to be used already registered?
•
If No, continue to step 6.
•
If Yes, continue to step 9.
In the Select Columns dialog, click New.
Result: The first New Column dialog opens.
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7 Run a method
7.3 Start a method run
Step
Action
7
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.
•
Press and hold the trigger to create a beam.
•
When the 2D Barcode scanner beeps, the column ID is registered and
displayed in the dialog.
•
Alternatively, manually enter the column ID, that you find on the column
label, in the dialog using your keyboard.
•
Click Continue.
Result: The expanded New Column dialog opens.
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7 Run a method
7.3 Start a method run
Step
Action
8
In the expanded New Column dialog:
•
Enter Alias (optional)
•
Select Technique and Column type from the drop-down lists.
•
Check the Set medium expiration date check box and select a date from
the drop-down list.
•
Click OK.
Tip:
Alias can be used for easy identification of a column.
Result: The entered information is saved and the dialog closes.
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7 Run a method
7.3 Start a method run
Step
Action
9
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
used in the run from the drop-down list.
•
Click OK.
Result: The run starts. All necessary actions occur automatically according
to the method, including ending of the run.
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7 Run a method
7.4 Monitor the run
7.4
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 customize 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).
1
2
3
116
4
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7 Run a method
7.4 Monitor the run
Part
Description
1
Run Data: Presents current run data values.
2
Chromatogram: Illustrates data as curves.
3
Run Log: Presents all registered actions.
4
Flow Scheme: Illustrates the current flow path.
Customize Run Data
The Run Data pane shows real-time data
from available monitors during the run.
To change what parameters to display
in the pane:
•
Click the Customize icon to open the
Customize dialog.
•
Under the Run Data Groups tab, select the parameters to display.
•
Click OK to close the dialog.
Customize Chromatogram
The Chromatogram pane displays a graphical presentation of registered data from
available monitors during the run.
To change the chromatogram properties:
•
Click the Customize icon to open the Customize dialog.
•
Make appropriate selections under the tabs described in the table below.
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117
7 Run a method
7.4 Monitor the run
•
Tab
Customization
Curves
Select curves to be displayed.
Curve style and color
Customize the appearance of the displayed curves.
X-axis
Select base unit (time or volume) and set X-axis scale.
Y-axis
Select which Y-axes to display and set Y-axis scale for
the different curves.
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
Y-axis scale refers to
click the curve of interest.
zoom in the curves
select the area of interest by moving the pointer with the
left mouse button held down.
Result: The Y-axis scale changes to the unit applicable for
the curve.
Result: The selected area is enlarged.
zoom out again
click the right mouse button and select Reset Zoom.
Result: The original area is restored in one step.
Customize Run Log
The Run Log pane displays all registered actions during the run. Scroll up to see the entire
log.
To change what items to display in the pane:
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7 Run a method
7.4 Monitor the run
•
Click the Customize icon to open the Customize dialog.
•
Under the Run Log tab, select the items to display.
•
Click OK to close the dialog.
Note:
These operations determine which data are shown in the Run Log pane during
a run. All original raw data are saved and can never be modified or removed
from the result.
Flow scheme
The Flow Scheme pane displays the current flow path during the run. Color indication
is applied, as shown in the table below. Real-time data from monitors are also displayed
in the flow scheme. See illustration below.
Color
Indication
Green
Open flow path
Grey
Closed flow path
Red (not shown)
Alarm
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7 Run a method
7.4 Monitor the run
120
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7 Run a method
7.5 After run procedures
7.5
After run procedures
Introduction
This section describes how to clean the instrument and columns after a chromatographic
run, and how to prepare the system for storage.
The instrument and the columns should be cleaned between the runs. This will prevent,
for example, sample contamination, protein precipitation and column clogging. If the
instrument is not going to be used for a couple of days or longer, the instrument, columns
and the pH flow cell should be filled with storage solution. For further information about
cleaning and maintenance procedures, see the User Manual.
Tip:
To clean and fill the instrument and columns with storage solution, use System
CIP and Column CIP either as separate, predefined methods or as phases 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:
•
Rinse the instrument with one or several cleaning solution(s) (e.g., NaOH, buffer solution or distilled water) using System CIP.
•
Empty the Fraction collector.
•
Clean all spills on the instrument and on the bench using a moist tissue.
•
Empty the waste vessel.
•
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:
•
Fill the system and inlets with storage solution (e.g., 20% ethanol) using System CIP.
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7 Run a method
7.5 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:
•
Fill the column with storage solution (e.g., 20% ethanol) using Column CIP.
pH electrode storage
Follow the instructions below to fill the pH flow cell with storage solution. The calibration
function is used to switch the position of the pH valve. However, no calibration is performed.
Step
Action
1
Open the System Control module and select System:Calibrate.
Result: The Calibration dialog opens.
2
In the Calibration dialog, select pH from the Monitor to calibrate drop-down
list.
3
Press
.
Result: The pH valve switches to the calibration position.
4
122
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.
Getting Started with ÄKTA avant 29-0351-83 AA
7 Run a method
7.5 After run procedures
Step
Action
5
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.
6
Press
.
Result: The pH valve switches back to the default position and the Calibration
dialog closes. No calibration is performed.
Log off or exit UNICORN
Follow the instruction below to log off or exit UNICORN. This can be performed from any
of the UNICORN modules.
If you want to...
then...
log off UNICORN
select File:Log off.
Result: All open UNICORN modules close and the Log On
dialog opens.
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7 Run a method
7.5 After run procedures
If you want to...
then...
exit UNICORN
select File: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.
Shut down the instrument
Switch off the instrument by pressing the Power switch to the O position.
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8 Evaluate and print the results
8
Evaluate and print the results
About this chapter
This chapter describes how to use the Evaluation module to evaluate and print the results
of a run. For further information, please refer to UNICORN 6 Evaluation Manual.
In this chapter
This chapter contains the following sections:
Section
See page
8.1 View the results
126
8.2 Peak integration
130
8.3 Print the results
135
Getting Started with ÄKTA avant 29-0351-83 AA
125
8 Evaluate and print the results
8.1 View the results
8.1
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.
2
Select the Results tab.
3
Select the result to open and click the Open a Result button in the navigator
toolbar.
Result: The result is opened in a Chromatogram pane. Available chromatograms and peak tables are displayed.
126
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8 Evaluate and print the results
8.1 View the results
Illustration of the Chromatogram
pane
The Chromatogram pane displays the curves from the method run.
Customize Chromatogram
To change the chromatogram properties:
•
Click the Customize icon to open the Customize dialog.
•
Make appropriate selections under the tabs described in the table below.
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127
8 Evaluate and print the results
8.1 View the results
•
Tab
Customization
Curves
Select curves to be displayed.
Curve style and color
Customize the appearance of the displayed curves.
X-axis
Select base unit (time, CV or volume) and set X-axis
scale.
Y-axis
Select which y-axes to display and set Y-axis scale for
the different curves.
Header
Select which parameters (variables, questions and/or
notes) to display in the header information at top of the
chromatogram.
Click OK to save the changes and close the dialog.
Note:
These operations determine which curves are displayed and their appearance
in the Chromatogram pane and on printouts. The original raw data curves
can never be modified or removed from the result.
View details in Chromatogram
Follow the instructions below to view selected parts of the chromatogram.
If you want to...
then...
identify a curve
position the mouse pointer over the curve of interest.
Result: The curve name is displayed.
select which curve the
Y-axis scale refers to
click the curve of interest.
zoom in on the curves
select the area of interest by moving the pointer with the
left mouse button held down.
Result: The Y-axis scale changes to the unit applicable for
the curve.
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.
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8 Evaluate and print the results
8.1 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.
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8 Evaluate and print the results
8.2 Peak integration
8.2
Peak integration
Introduction
Peak integration is used to identify and measure curve characteristics, including peak
areas, retention times and peak widths. This section describes how to perform peak 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.
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8 Evaluate and print the results
8.2 Peak integration
Step
Action
3
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.
•
Click OK.
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.
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8 Evaluate and print the results
8.2 Peak integration
Illustration of Chromatogram
and Peak Table
The illustration below shows a Chromatogram pane including a Peak Table.
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.
•
Under the Peak Table tab:
-
132
Select which data to display in the peak tables from the Select peak table
columns list.
Getting Started with ÄKTA avant 29-0351-83 AA
8 Evaluate and print the results
8.2 Peak integration
-
Click OK to display the settings and close the dialog.
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).
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8 Evaluate and print the results
8.2 Peak integration
Save results
Follow the instruction below to save your changes.
134
If you want to...
then...
save your changes to
the original result
click the Save icon in the toolbar.
save the changed result
as a new result
•
select File:Save As to open the Save Result As dialog.
•
In the Save Result As dialog:
-
select the location for the new result
-
write a name for the new result
-
click Save.
Getting Started with ÄKTA avant 29-0351-83 AA
8 Evaluate and print the results
8.3 Print the results
8.3
Print the results
Introduction
This section describes how to print a chromatogram and a standard format report.
UNICORN uses the printers and printer settings that are installed on your computer.
Customize the view
Before printing, make sure that the view is adjusted to show what is needed in the report/printout. Customize the chromatogram and/or peak table and view details in the
chromatogram according to the instructions above. Before printing, check that:
•
correct curves are displayed
•
correct data are displayed in the peak table
•
a suitable scaling of the axes has been selected
•
a suitable zoom factor has been selected
Print a chromatogram and a
peak table
Follow the instruction below to print a chromatogram and, if peak integration has been
performed, the related peak table.
Step
Action
1
Open the Evaluation module and open a result.
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135
8 Evaluate and print the results
8.3 Print the results
Step
Action
2
Click the Print icon.
Result: The Print Chromatograms dialog opens.
3
In the Print Chromatograms dialog:
•
Select Printer and Print Format.
•
Click the Preview button.
Result: A preview of the chromatogram displays in the Customize Report
window.
4
136
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.
Getting Started with ÄKTA avant 29-0351-83 AA
8 Evaluate and print the results
8.3 Print the results
Step
Action
5
In the Customize Report window, click the Print icon.
Result: The Print dialog opens.
6
In the Print dialog, select Printer from the list and click OK.
Result: Your chromatogram is printed.
Print a report
Follow the instruction below to print a standard format report.
Step
Action
1
Open the Evaluation module and open a result.
Getting Started with ÄKTA avant 29-0351-83 AA
137
8 Evaluate and print the results
8.3 Print the results
Step
Action
2
Click the Report icon.
Result: The Generate Report dialog opens.
3
In the Generate Report dialog:
•
Select any of the predefined formats from the list.
•
Click the Preview button.
Result: A preview of the report displays in the Customize Report window.
4
138
Are you satisfied with the layout?
•
If Yes, continue to step 5.
•
If No, select File:Exit to return to the Generate Report dialog and choose
another format.
Getting Started with ÄKTA avant 29-0351-83 AA
8 Evaluate and print the results
8.3 Print the results
Step
Action
5
In the Customize Report dialog, click the Print icon.
Result: The Print dialog opens.
6
In the Print dialog, select Printer from the list and click OK.
Result: Your report is printed.
Getting Started with ÄKTA avant 29-0351-83 AA
139
9 Maintenance
9
Maintenance
Introduction
This chapter provides schedules for preventive maintenance that should be performed
by the user of ÄKTA avant. Regular maintenance is essential for reliable function and
results. Refer to the ÄKTA avant User Manual for detailed instructions.
WARNING
Always use appropriate personal protective equipment during operation and maintenance of ÄKTA avant system.
Maintenance program
An overview of the preventive maintenance to be performed on ÄKTA avant is outlined
below. See the ÄKTA avant User Manual for detailed information about the maintenance
procedures.
Maintenance is divided into:
•
Daily maintenance
•
Weekly maintenance
•
Monthly maintenance
•
Bi-annual maintenance
•
Maintenance when required
WARNING
Electrical shock hazard. All repairs should be done by service
personnel authorized by GE Healthcare. Do not open any covers
or replace parts unless specifically stated in the user documentation.
140
Getting Started with ÄKTA avant 29-0351-83 AA
9 Maintenance
Periodic maintenance program
The following periodic maintenance should be performed by the user of ÄKTA avant.
Interval
Maintenance action
Daily
Calibrate the pH monitor
Weekly
Calibrate pressure monitors
Weekly
Change pump rinsing solution
Weekly
Replace online filter in the Mixer
Monthly
Check the Flow restrictor
Bi-annual
Clean the UV flow cell
Bi-annual
Replace pH electrode
Maintenance when required
The following maintenance should be performed by the user of ÄKTA avant when required.
Maintenance action
Clean the instrument externally
Perform System CIP
Perform Column CIP
Clean the Fraction collector
Replace tubing and connectors
Storage of pH electrode
Clean the pH electrode
Clean the Conductivity flow cell
Calibrate the Conductivity monitor
Calibrate the UV monitor
Replace Mixer
Getting Started with ÄKTA avant 29-0351-83 AA
141
9 Maintenance
Maintenance action
Replace o-ring in Mixer
Replace the UV flow cell
Replace the Flow restrictor
Replace inlet filters
Clean the check valves
Replace check valves
Replace pump piston seals
Replace pump pistons
Replace pump rinsing system tubing
Replace valve modules
142
Getting Started with ÄKTA avant 29-0351-83 AA
For local office contact information, visit
www.gelifesciences.com/contact
GE, imagination at work and GE monogram are trademarks of General Electric
Company.
GE Healthcare Bio-Sciences AB
Björkgatan 30
751 84 Uppsala
Sweden
Windows is a trademark of Microsoft Corporation.
www.gelifesciences.com/AKTA
ÄKTA, AxiChrom and UNICORN are trademarks of GE Healthcare companies.
© 2009-2012 General Electric Company – All rights reserved.
First published Dec. 2012
All goods and services are sold subject to the terms and conditions of sale of
the company within GE Healthcare which supplies them. A copy of these terms
and conditions is available on request. Contact your local GE Healthcare representative for the most current information.
UNICORN: Any use of this software is subject to GE Healthcare Standard Software
End-User License Agreement for Life Sciences Software Products. A copy of this
Standard Software End-User License Agreement is available on request.
GE Healthcare Europe GmbH
Munzinger Strasse 5, D-79111 Freiburg, Germany
GE Healthcare UK Limited
Amersham Place, Little Chalfont, Buckinghamshire, HP7 9NA, UK
GE Healthcare Bio-Sciences Corp.
800 Centennial Avenue, P.O. Box 1327, Piscataway, NJ 08855-1327, USA
GE Healthcare Japan Corporation
Sanken Bldg. 3-25-1, Hyakunincho Shinjuku-ku, Tokyo 169-0073, Japan
imagination at work
29-0351-83 AA 03/2013
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