GE AKTA START (01) PDF MANUAL

FREE ENGLISH GE AKTA START (01) PDF USER GUIDE

FREE ENGLISH GE AKTA START (01) PDF USER MANUAL

FREE ENGLISH GE AKTA START (01) PDF OWNER GUIDE

FREE ENGLISH GE AKTA START (01) PDF OWNER MANUAL

FREE ENGLISH GE AKTA START (01) PDF REFERENCE GUIDE

FREE ENGLISH GE AKTA START (01) PDF INSTRUCTION GUIDE

FREE ENGLISH GE AKTA START (01) PDF REFERENCE MANUAL

FREE ENGLISH GE AKTA START (01) PDF INSTRUCTION MANUAL

FREE ENGLISH GE AKTA START (01) PDF OPERATING INSTRUCTIONS

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GE AKTA START (01) PDF SUMMARY:

What is the purpose of the Operating Instructions? The Operating Instructions provide the instructions needed to install, operate and maintain the ÄKTA start system in a safe way.

What should users do before operating the product? All users must read the entire Operating Instructions before installing, operating or maintaining the product. Always keep the Operating Instructions at hand when operating the product. Do not operate the product in any other way than described in the user documentation.

What is ÄKTA start intended for? ÄKTA start is a liquid chromatography system used for preparative purification of proteins at laboratory-scale. The system can be used for a variety of research purposes to fulfill the needs of the users in academia and the life sciences industry. ÄKTA start is intended for research use only, and shall not be used in any clinical procedures, or for diagnostic purposes.

What are the prerequisites for using the system? In order to follow the manual and use the system in the manner it is intended, it is important that you understand the concepts of liquid chromatography and that you have read and understood the Safety instructions chapter in the Operating Instructions.

What do the following terms mean: WARNING, CAUTION, and NOTICE?

WARNING indicates a hazardous situation which, if not avoided, could result in death or serious injury.
CAUTION indicates a hazardous situation which, if not avoided, could result in minor or moderate injury.
NOTICE indicates instructions that must be followed to avoid damage to the product or other equipment.

What are some general safety precautions to follow when using ÄKTA start?

Do not operate ÄKTA start in any other way than described in the ÄKTA start user documentation.
Operation and user maintenance of ÄKTA start should be performed according to the instructions described in ÄKTA start Operating Instructions and ÄKTA start Maintenance Manual.
Do not use any accessories not supplied or recommended by GE.
Do not use ÄKTA start if it is not working properly, or if it has suffered any damage, for example: damage to the power cord, its plug or the Frac30 cable; damage caused by dropping the equipment; damage caused by splashing liquid onto it.

What precautions should be followed when using flammable liquids with ÄKTA start?

Fire Hazard. Before starting the system, make sure that there is 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.
The products are not approved for work in a potentially explosive atmosphere and do not fulfill the requirements of the ATEX Directive.

What should be done with the 24% ethanol that ÄKTA start is filled with at delivery? ÄKTA start is filled with 24% ethanol at delivery, and this alcohol can be hazardous to humans if consumed. The alcohol must be flushed out before assembling, testing, or integrating ÄKTA start into the intended process context.

What personal protection measures should be taken when working with ÄKTA start?

Always use appropriate personal protective equipment during operation and maintenance of ÄKTA start.
When using ÄKTA start, use personal protective equipment like goggles, lab coat, protective shoes, and gloves to avoid any circumstances of spillage.
The tubing cutter is very sharp and must be handled with care to avoid injuries.
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.
The operator has to take all necessary actions to avoid spreading hazardous biological agents in the vicinity of the equipment. The facility should comply with the national code of practice for biosafety.

What precautions should be taken when installing or moving the instrument?

Moving the instrument horizontally: one person is 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.
Power cord: Only use grounded power cords delivered or approved by GE.
Access to power switch and power cord with plug: Do not block access to the power switch and power cord. The power switch must always be easy to access. The power cord with plug must always be easy to disconnect.
Installing the computer (optional): The computer should be installed and used according to the instructions provided by the manufacturer of the computer.
Disconnect power: Always switch off power to ÄKTA start before an instrument module is removed or installed, or a cable is connected or disconnected.
Protective ground: ÄKTA start must always be connected to a grounded power outlet.
Vents on ÄKTA start: To ensure adequate ventilation, keep papers and other objects away from the vents of the instrument.
Any computer used with the equipment shall comply with IEC 60950 and be installed and used according to the manufacturer’s instructions.
Frac30 should not be connected or disconnected from ÄKTA start when the instrument is powered ON.

What are some precautions to take during system operation?

Rotating the instrument: Make sure that there is always at least 20 cm of free space around ÄKTA start to allow for sufficient ventilation. When turning or moving 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 an increase in pressure, or block liquid flow. To avoid the risk of knocking over bottles, always place bottles on the buffer tray and turn or move carefully.
Hazardous chemicals during run: When using hazardous chemicals, run the System cleaning template to clean and flush the entire system tubing with distilled water, before service and maintenance.
Setting: Check that the correct outlet size settings are used. Make sure that tubing and fittings are properly connected and secured. Make sure that the pressure limit settings are correct before starting the run.
Max weight on Buffer tray: Do not place containers with a volume of more than 1 liter each on the Buffer tray. The maximum allowed weight on the Buffer tray is 5 kg.
Large spillage: Switch off ÄKTA start and unplug the power cord, if large spillage occurs.
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.
Prefill UV flow cell: Make sure that the UV flow cell is prefilled with liquid before starting the system.
Avoid condensation: If ÄKTA start is kept in a cold room, cold cabinet or similar, keep the instrument switched on in order to avoid condensation.
Avoid overheating: If ÄKTA start is kept in a cold cabinet and the cold cabinet is switched off, make sure to switch off the instrument and keep the cold cabinet open to avoid overheating.
Place the computer in room temperature: If ÄKTA start is placed in a cold room, place the computer outside the cold room and use the PC Connectivity cable delivered with the instrument to connect to the computer.
Keep the pump cover open when not using the system: Open the peristaltic pump cover after you switch off the equipment to enhance the life time of the pump tubing.

What are some precautions to take when performing maintenance on ÄKTA start?

Electrical shock hazard: Do not open any covers or parts unless specified in the user documentation. Except for the maintenance and service described in the user documentation, all other repairs should be done by service personnel authorized by GE.
Only spare parts and accessories that are approved or supplied by GE may be used for maintaining or servicing ÄKTA start.
Disconnect power: Always switch off power to the instrument before replacing any component on the instrument or cleaning the instrument, unless stated otherwise in the user documentation.
Spillage Hazard: Avoid spillage of fluids on the surfaces of the instrument which have cables, plugs, and other wirings. Be careful if there is spillage of fluids on the tray while trying to remove the tray from ÄKTA start.
NaOH is corrosive and therefore dangerous to health: When using hazardous chemicals, avoid spillage and wear protective glasses and other suitable Personal Protective Equipment (PPE).
Hazardous UV light: Always switch off power to the instrument before replacing the UV flow cell.
If hazardous chemicals are used for system or column cleaning, wash the system or column with a neutral solution in the last phase or step before maintenance.
Cleaning: Keep the instrument dry and clean. Wipe regularly with a soft damp tissue and, if necessary, a mild cleaning agent. Let the instrument dry completely before use.

What do the following labels on the instrument mean?

Do not open any covers on the instrument. This will void the warranty.
Keep the pump cover open when not using the system. Open the pump cover after you switch off the instrument.
Warning! Consult the Operating Instructions before using the system.
Pinch hazard. Switch off the Pump before loading tubing.

What do the symbols on the instrument labels mean?

Warning! Read the user documentation before using the system. Do not open any covers or replace parts unless specifically stated in the user documentation.
This symbol indicates that the waste of 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 contains hazardous materials in excess of the limits established by the Chinese standard GB/T 26572 Requirements of concentration limits for certain hazardous substances in electrical and electronic products.
The system complies with applicable European directives.
The equipment complies with the requirements for electromagnetic compliance (EMC) in Australia and New Zealand.
Eurasian Conformity mark: the single conformity mark indicates that the product is approved for circulation on the markets of the member states of the Eurasian Customs Union.
This symbol indicates that ÄKTA start has been certified by a Nationally Recognized Testing Laboratory (NRTL). This product Conforms to UL 61010-1, and is Certified to CAN/CSA-C22.2 No. 61010-1.
Degree of protection provided by the enclosure.
Year (YYYY) and month (MM) of manufacture.

How do you perform an emergency shutdown? Switch off power to the instrument by pressing the power switch to the O position or by disconnecting the power cord from the instrument. The run is interrupted immediately.

What happens in the event of a power failure to ÄKTA start?

The run is interrupted immediately.
The data collected up to the time of the power failure is saved in UNICORN start (if the system is connected to a computer), or on the USB memory stick.
On the ÄKTA start Instrument Display, all four touch buttons will be highlighted.

What happens in the event of a power failure to UNICORN start on a computer?

The computer with UNICORN start installed shuts down.
The run is interrupted immediately.
Data generated up to 10 seconds before the power failure can be recovered.

What must be done with the product before decommissioning? The product must be decontaminated before decommissioning. All local regulations must be followed with regard to scrapping of the equipment.

How should the product be disposed of? When taking the product out of service, the different materials must be separated and recycled according to national and local environmental regulations. Waste 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 the equipment.

What is the function of the Instrument Display? The Instrument Display enables the user to control the system by selecting the intended operation: Start a run and control an ongoing run; view the progress of the ongoing run; manage user defined methods; and perform maintenance and service.

What should you not use to operate the Instrument Display? Do not operate the Instrument Display with a sharp and hard object.

What are the different functionality menu options on the ÄKTA start home screen?

Method run: Use a quick start or a method template to perform a run.
Manual run: Perform a run by providing parameters manually.
Create method: Create, edit, import and delete user methods.
Settings and service: Configure settings and calibrate modules, perform diagnostics tests.

What are the functions of the touch buttons on the Instrument Display?

Help: Opens a new dialog screen, providing information about the content of the current screen or indicates where further information or instructions can be found.
Home: Opens the ÄKTA start home screen.
Forward: Opens the next screen in the current workflow.
Return: Opens the previous screen in the current workflow.
Increment (up arrow): The value in the text field can be increased or decreased by tapping up or down arrow.
Decrement (down arrow): Tapping the value opens the numpad and a new value can be typed in.
Next: Opens the next screen.
Back: Returns to the previous screen.
Run: Starts a run.
Pause: Pauses the ongoing run by stopping the Pump. The flow rate settings and the gradient values are retained.
Continue: Continues a run that has been paused.
Hold: Holds an ongoing run, with current flow rate, valve positions, and with set %B concentration. The gradient is held at the value displayed.
Resume: Resumes a run that is put on hold.
Edit run: Opens a new screen for editing the current run parameters.
Execute: Executes the edited run parameters during a run.
OK: Confirms a selection or an action.
Cancel: Cancels a selection or an action.

What does the End button do?

Terminates the ongoing run; always followed by a screen that requires confirmation of the action. When a run is completed, it closes the application and returns to the ÄKTA start home screen.

What does the Exit button do?

Opens the graphic view of the run in progress, displaying the plot of the UV absorbance (mAU) versus Time (min).

What does the Graph icon do?

Saves a user method that was either created or edited.

What does the Save button do?

Confirms and initiates a run from a specific template or user method.

What does the Select button do?

Creates a user method based on selected template. The run parameters have to be edited as needed.

What does the Create button do?

Confirms an action.

What does the No button do?

Rejects an action.

How do I access the Instrument Display Help?

The ÄKTA start Instrument Display Help is accessible from every screen on the Instrument Display, by tapping the question mark in the upper right corner. The Help text provides information about the content of the current screen or refers to more detailed documentation.

What options are available under Method run?

The display option Method run allows the user to run methods based on Quick start techniques or predefined method templates, user created methods, and predefined methods such as PumpWash, and System cleaning.

What is displayed when Method run is selected?

When Method run is selected, further options are displayed in the Method run screen.

What are the method types available under Method run?

Quick start
Templates
User defined
Prepare system

What do the Quick start templates display?

Displays the Quick start templates available with ÄKTA start. For description of the Quick start methods, see Section 6.4.2 Quick start, on page 161.

What do the Templates display?

Displays the method templates for chromatography techniques available with ÄKTA start. For a description of the templates, see Section 6.4.3 Templates, on page 167.

What does User defined display?

Displays the methods created by the user. For detailed instructions on creating and managing user defined methods, see Section 6.6 Manage methods and files, on page 183.

What does Prepare system display?

Displays the predefined system methods that are used to clean the system flow path. For detailed instructions on running system methods, see Section 6.4.5 Prepare system methods, on page 177.

What options are available under Create method?

The display option Create method allows the user to create new methods, edit or delete existing user methods and also import methods stored on a USB memory stick connected to the instrument.

What is displayed when Create method is selected?

When Create method is selected, further options are displayed in the Create method screen.

What operations are available under Create method?

Create
Edit
USB Import
Delete

What does the Create option display?

Displays the method templates that can be used to create a new method.

What does the Edit option display?

Displays the user methods stored on the instrument that can be edited, if required.

What does the USB Import option display?

Displays a list of user methods stored on a USB memory stick that can be imported to the instrument.

What does the Delete option display?

Displays the user methods stored on the instrument that can be deleted, if required.

What options are available under Settings and service?

The display option Settings and service allows the user to set the delay volume and perform maintenance, calibration, diagnostics, and troubleshooting of the modules on the wet side of the instrument.

What modules are covered in Settings and service – Screen 1?

Fraction collector
Pressure sensor
Pump
System

What modules are covered in Settings and service – Screen 2?

Buffer valve
Sample valve
Wash valve
Outlet valve

What modules are covered in Settings and service – Screen 3?

Main board
UV
Display
Conductivity

What are the functions for the Fraction collector module in Settings and service – Screen 1?

Enable Frac/Disable Frac: Activates/Deactivates the Fraction collector connection.
Diagnostics: Performs a feed tube test that checks if the Bowl assembly rotates correctly. Performs a Home test to verify that the Bowl assembly returns to Home position.
Run log: Shows the number of hours the Frac drive has been active and allows the number of hours of drive usage to be reset.

What are the functions for the Pressure sensor module in Settings and service – Screen 1?

P set: Shows the pressure in the flow path.
Zero offset: Sets the pressure reading to zero. The Pump has to be off and the flow path connection to the Pressure sensor has to be open.

What are the functions for the Pump module in Settings and service – Screen 1?

Calibration: Calibrates the Pump, so that it delivers liquid with an accurate flow rate.
Pump tubing log: Shows the number of hours the pump tubing has been in use (Tubing run) and allows the tubing usage count to be reset (after the pump tubing is replaced). Shows the number of hours the Pump has been in use (Pump run) and allows the Pump usage count to be reset (after the Pump is replaced).
Diagnostics: Adjusts the flow rate and start/stop the flow (Pump).

What are the functions for the System module in Settings and service – Screen 1?

Delay volume setting: Sets the delay volume. The delay volume represents the volume of the liquid in the flow path between the UV and the collection tubes.
Firmware update: Updates the system firmware.
Switch valve timing: Sets the Switch valve timing. It is recommended to optimize the timing of switch valve (Buffer valve) when wavy gradients are obtained or when fluctuations in the step gradient are observed during either system performance tests or chromatography runs.
Export system report to USB: Exports system error logs to a USB memory stick in a text format. A system error report is useful when trying to understand a possible error that requires remote service support.

What are the functions for the Buffer valve module in Settings and service – Screen 2?

Valve position: Shows which inlet connection in the valve is open, Buffer A or Buffer B.
Turn valve: Switches the valve between its two positions i.e., the A and B inlet ports.
Valve switches: Displays the number of counts the valve has switched. Reset: Resets the counts to zero when a valve has been replaced.

What are the functions for the Sample valve module in Settings and service – Screen 2?

Valve position: Shows which inlet connection in the valve is open, Buffer or Sample.
Turn valve: Switches the valve between its two positions i.e., the Buffer and Sample inlet ports.
Valve switches: Displays the number of counts the valve has switched. Reset: Resets the counts to zero when a valve has been replaced.

What are the functions for the Wash valve module in Settings and service – Screen 2?

Valve position: Shows which outlet connection in the valve is open, Waste or Column.
Turn valve: Switches the valve between its two positions i.e., the Waste and Column outlet ports.
Valve switches: Displays the number of counts the valve has switched. Reset: Resets the counts to zero when a valve has been replaced.

What are the functions for the Outlet valve module in Settings and service – Screen 2?

Valve position: Shows which outlet connection in the valve is open, Waste or Collection.
Turn valve: Switches the valve between its two positions i.e., the Waste and Collection/Frac outlet ports.
Valve switches: Displays the number of counts the valve has switched. Reset: Resets the counts to zero when a valve has been replaced.

What are the functions for the Main board module in Settings and service – Screen 3?

Power supply: Shows the actual voltage.
Memory test: Performs a memory test.
Temperature: Shows the temperature of the main board.
FPGA test: Performs a test of the FPGA hardware.

What are the functions for the UV module in Settings and service – Screen 3?

UV LED calibration: Calibrates the UV LED intensity.
Flow cell path length: Derives the actual path length of the flow cell.
Diagnostics: Troubleshoots the UV module.
Configuration: Configures a new UV monitor and flow cell.

What are the functions for the Display module in Settings and service – Screen 3?

Touch screen calibration: Calibrates the touch screen.
Color test: Performs a diagnostics test for the colors displayed on the touch screen.
Diagnostics: Performs a diagnostics test to check the backlight performance of the display.
Log book: Shows the number of hours the instrument display has been in use.

What are the functions for the Conductivity module in Settings and service – Screen 3?

Calibration: Calibrates the Conductivity flow cell and the temperature sensor.
Configuration: Sets a new reference temperature value, usually after replacement of the Conductivity flow cell.
Advanced calibration: Calibrates the temperature of the Conductivity flow cell.

What are the space requirements for the ÄKTA start?

The minimum recommended space for ÄKTA start is a benchtop space with dimensions of 100 cm in width and 80 cm in depth, with a clearance of 34 cm in height.

What are the equipment dimensions for ÄKTA start and Frac30?

ÄKTA start has dimensions of 360 mm in width, 280 mm in depth, and 340 mm in height. Frac30 has dimensions of 285 mm in width, 280 mm in depth, and 270 mm in height.

What is the weight of ÄKTA start and Frac30 with packaging?

ÄKTA start (with packaging) weighs 12 kg and Frac30 (with packaging) weighs 6 kg.

How should the delivery boxes for ÄKTA start and Frac30 be transported?

To transport the delivery boxes containing the instrument and Fraction collector, use a hand truck suitable for lightweight packages. Each box can be lifted by 1 person without the help of any lifting equipment.

What should be done with the original packing material?

Save all the original packing material. If the system has to be repacked, for transportation or otherwise, it is important that the system can be safely packed using the original packing material.

What is the caution about ÄKTA start upon delivery?

ÄKTA start is filled with 24% ethanol at delivery. The alcohol can be hazardous to humans if consumed. Flush out the alcohol before assembling, testing or integrating ÄKTA start into the intended process context.

How should I unpack ÄKTA start?

Open the delivery box by cutting the adhesive tape at the top of the box.
Take out the document placed at the top of the package and read the Unpacking Instructions.
Take out the box placed at the top of the package. The box contains the accessories delivered with the instrument.
Hold the red strap, and then lift the instrument out of the delivery box.
Open the strap lock and remove the strap.
Remove the foam cushion from the top of the instrument.
Remove the foam cushion from the bottom of the instrument by carefully lifting the instrument.
Remove the plastic bag by gently tilting the system back and forth while pulling out of the plastic bag.

How should I unpack Frac30?

Open the Frac30 delivery box by cutting the adhesive tape at the top of the box.
Take out the document placed at the top of the package and read the Unpacking Instructions.
Holding the red strap, lift the fraction collector out of the delivery box. Place the Fraction collector on the laboratory bench.
Open the strap lock and remove the strap.
Remove the foam cushion from the top of the Fraction collector.
Remove the foam cushion from the bottom of the fraction collector by carefully lifting the Fraction collector.
Remove the plastic bag.
Remove the Bowl assembly from the Base unit: Gently move the Dispenser arm counterclockwise to the end position. Pull the Drive assembly outwards and hold it at the retracted position. At the same time, lift the Bowl assembly.
Remove the foam cushion located on the Base unit.
Re-assemble the Bowl assembly on to the Base unit: Orient the Bowl to match the aligning groove and the aligning features located on the bowl holder. Slightly push the Drive assembly laterally and lower the Bowl assembly onto the Base unit.

What should I not do when unpacking Frac30?

Never lift the Frac30 by the Dispenser arm. This may damage the Fraction collector. Do not damage or break the warranty seal label during unpacking of Frac30.

What is included in the accessories package?

Unpacking Instructions
System certificate
Operating Instructions
Maintenance Cue Card
System Cue Card
CD containing user documentation files
EU Declaration Of Conformity
Instructions: ÄKTA start UV Module and Support Information
Accessories box

What actions must be performed to install ÄKTA start?

Install pump tubing
Connect power supply to ÄKTA start
Connect Frac30 to ÄKTA start
Perform initial setup of the UV monitor if required, as described in the ÄKTA start UV Module and Support Information
(Optional) Connect ÄKTA start to the UNICORN start computer

What warnings should be considered when installing ÄKTA start?

Only use grounded power cords delivered or approved by GE. Make sure that the supply voltage at the wall outlet corresponds to the marking on the instrument, before connecting the power cord. ÄKTA start must always be connected to a grounded power outlet.

How do I install the pump tubing?

Open the top cover fully.
Place the tubing between the rollers and the track, press the tubing against the pump head inner wall.
Lower the top cover until it clicks into its fully closed position.

How do I connect the power to ÄKTA start?

Select the correct power cord to be used. ÄKTA start is delivered with 2 alternative power cords: Power cord with US-plug, 2 m and Power cord with EU-plug, 2 m.
Connect the power cord to the Power input connector on the left side of the instrument and to a grounded wall outlet 100-240 VAC, 50/60 Hz.

How do I connect Frac30 to ÄKTA start?

Frac30 should not be connected or disconnected from ÄKTA start when the instrument is powered ON.

Connect the Frac30 cable between the ports on the back of the Fraction collector and the instrument.
Switch on ÄKTA start.
Enable the connection to Frac30 from the Instrument Display: In the ÄKTA start home screen, tap Settings and service. The Settings and service Screen 1 opens. In the Settings and service screen, tap Fraction collector. The Fraction collector screen opens. In the Fraction collector screen, tap Enable Frac to enable the connection of the Fraction collector.

How do I connect a computer to ÄKTA start?

Before connecting the computer to ÄKTA start, install the UNICORN start software on the computer.

Connect power to the computer and monitor, and then switch on the computer and ÄKTA start. The instrument displays the ÄKTA start home screen.
Connect the PC Connection Cable between the connector marked as PC Connection on the back of ÄKTA start and a USB port on the computer.
Launch UNICORN start and connect to ÄKTA start.

What should I do before starting the run?

Make sure that the system connection is established before starting the run. Always make sure to be in the ÄKTA start home screen (connected state) when trying to connect from the System Control module.

What does the ÄKTA start flow path contain?

The flow path contains Pump, Mixer, Valves, and UV, Conductivity and Pressure monitors.

What are the modules in the ÄKTA start flow path?

Buffer valve
Mixer
Sample valve
Pump
Pressure sensor
Wash valve
Injection valve (manual)
Column
UV Monitor
Conductivity Monitor
Outlet valve
Fraction collector

What is the state of the flow path upon delivery?

ÄKTA start is delivered with the entire flow path assembled and pre-filled with storage solution (24% ethanol).

What tubing is connected to the instrument?

The following table lists the tubing connected to the instrument:

Tube connection	Module
Port I (Buffer A)	Buffer valve
Port II	Mixer
Port III (Buffer B)	Buffer valve
Port I (Sample)	Sample valve
Port II	Pump
Port III	Mixer
Port I (Waste)	Wash valve
Port II	Pressure sensor
Port III	Injection valve
Port 1	Injection valve
Ports 2 and 5	Injection valve
Port 3	Injection valve
Port 4 (Waste)	Injection valve
Port 6	Wash valve
Port I (Waste)	Outlet valve
Port II	Conductivity Monitor
Port III (Collection)	Outlet valve

Where should buffer bottles, sample bottles, and waste bottles be placed?

Buffer bottles are placed in the Buffer tray on top of the instrument. Sample bottle or tube may be placed on the bench on the left side of the instrument. A waste bottle may be placed on the bench on the right side of the instrument.

What is the maximum weight that the Buffer tray can hold?

Do not place bottles with a volume of more than 1 liter each on the Buffer tray. The total allowed weight on the Buffer tray is 5 kg.

What should I do before using the system for the first time?

Clean the flow path as described in Section 8.3 Cleaning the system flow path, on page 210.

How do I switch on the instrument?

Switch on the instrument by pressing the Power switch to the l position.
The instrument starts and initializes the display, showing the ÄKTA start home screen.

When should I calibrate the instrument?

The instrument is pre-calibrated at delivery, therefore no calibration is required when the instrument is installed. However, if the System performance test fails it is recommended to calibrate the modules.

When should I calibrate the Instrument Display?

If there are any issues with the response of the touch screen.

When should I calibrate the Pressure sensor?

If the pressure is outside the range of ±0.03 MPa, perform Zero offset.

When should I calibrate the Pump?

When chromatography run conditions are changed, e.g., viscosity of sample or buffer, temperature, back-pressure. The Pump and pump tubing requires calibration regularly. Recommended: once a week. After the pump tubing has been replaced with new tubing.

What is an important reminder about the pump tubing?

Do not leave the pump tubing inside the Pump when the Pump is not running.

When should I calibrate the UV Monitor?

When the signal is unstable, or readings appear to be incorrect. After cleaning, or after replacing the UV flow cell. When error/warning is seen on power ON. When a Baseline ignored message is shown with a clean UV flow cell. Before and after performing runs in the cold room.

When should I calibrate the Conductivity Monitor?

When the signal is unstable, or readings appear to be incorrect. After replacing the Conductivity flow cell.

What is the System performance method used for?

The System performance method is performed to make sure that the system is performing within acceptable limits. It is recommended to run the test at the time of installation of the instrument or after replacement of modules such as Pump, UV, Conductivity or Valves. System performance method can also be used at any time to check the condition of the system, for example, after a prolonged storage of the system.

Where can the System performance method be performed from?

The System performance method can be performed from both the Instrument Display and UNICORN start.

What should be done before starting the System performance method?

Calibrate all the modules before starting the test.
Make sure that there is no column connected.
It is recommended not to edit any run parameters during a test in order to avoid failure of the test.

What solutions are required for the System performance method?

Buffer A – DM water
Buffer B – 1.0% acetone, 1.0 M NaCl
Sample – 1.0% acetone, 1.0 M NaCl (Buffer B)

What checklist should be completed before starting a System performance method?

Calibration of all the modules: Pressure sensor, Pump, UV and Conductivity.
Make sure there is no column in the flow path.
Set the conductivity reference temperature to 20°C, save and enable the function.
Immerse Buffer port A inlet in Buffer A (DM water).
Immerse Buffer port B inlet in Buffer B (1.0% acetone, 1.0 M NaCl).
Immerse Sample valve inlet in sample (1.0% acetone, 1.0 M NaCl).
Make sure that 1 ml Sample loop is filled with sample (1.0% acetone, 1.0 M NaCl).
Make sure that 2 m of 0.5 mm ID tubing is connected to the Outlet valve at Waste position.
When performing System performance method without Fraction collector, make sure the Outlet valve fractionation tubing is inserted into a pre-weighed beaker.
When performing System performance method with Fraction collector, make sure the Outlet valve fractionation tubing is connected to Fraction collector with at least 5 pre-weighed tubes.
Make sure that the system is prefilled with DM water.
Take note of all the required observations by recording parameters while the System performance method is in progress. Enter the observed values in the System report template provided in Section 11.1 System Performance Report, on page 269.

How do I initiate the System performance method from the Instrument Display?

From the ÄKTA start home screen, tap Method Run.
In the Method run screen, tap Prepare system.
Select System performance method. To initiate the method, tap Select.
Tick the check box in order to save the results on a USB memory stick, and then tap Run to start the System Performance test.
Tap Exit to close the screen when the System performance method is completed.

What is the recommendation about editing parameters during the System performance method?

It is recommended not to edit any run parameters during a run to avoid failure of the test.

Can the System performance method run be ended before it is completed?

If required, the System performance method run can be ended before it is completed. Tap End to abort the test.

What should I do before unplugging the USB memory stick?

Do not unplug the USB memory stick until the Exit screen is displayed.

What are the acceptance criteria for the System performance method?

Activity	Time (min)	Approved interval
Pump wash	0	Mobile phase out through Waste
Back pressure	1	≤ 0.05 MPa at 1 ml/min, 0% B, flow through Outlet valve, Waste position
Repeat UV Auto zero	2
Back pressure	3	0.06 to 0.2 MPa at 5 ml/min
UV level	4	± 10 mAU
Conductivity level	4	± 1 mS/cm
Max. UV level	7	300 to 380 mAU at 1 ml/min, Sample valve, Sample position
Max. Conductivity level	7	65 to 95 mS/cm at 1 ml/min, Sample valve, Buffer position
Max. UV level	10	300 to 380 mAU; Request switch Injection valve to Inject position.
Max. Conductivity level	10	65 to 95 mS/cm
Request switch Injection valve back to Load position	13
Start gradient, 0% to 100% B in 10 minutes, start fractionation/collection	15

What are the approved interval check activity times for weighing fractions?

The approved interval check activity times are 0.8 to 1.2 g for weighing fractions no. 2, 3 and 4. The maximum difference between fractions is 0.1 g, and the approved time for weighing a beaker at the end of collection is 4.2 to 5.8 g.

What do you do if the System performance method fails?

If the System performance method fails, you should analyze the cause of the failure based on the acceptance criteria. Then, take the following actions: Re-calibrate the failed module, use buffer with proper composition, clean the failed module or entire system (refer to Chapter 8 Maintenance for more details on cleaning), and carefully follow the test instructions. Repeat the System performance method until it passes. If wavy or fluctuating gradient is observed, perform switch valve timing optimization. If the test fails after following the above actions, replace the failed module.

How do you start the System performance method from UNICORN start?

To start the test, go to System control: System: Performance Test and Report in UNICORN start. Select the method based on the fraction collector configuration: Use “Performance method with Frac” when the fraction collector is enabled, and use “Performance method without Frac” when the fraction collector is disabled. Read the method notes before starting the run and make a note of the result file location, then run the System performance method. The test report will state whether the method passed or failed.

What should you verify manually during the System performance test?

Manually verify pressure limits, fractionation/collection volumes, gradient levels, and all connections for leakage during the test, using the acceptance criteria.

What is the back pressure limit during the System performance test?

The back pressure limit is ≤ 0.05 MPa at 1 ml/min, 0% B, flow through Outlet valve, Waste position.

What are the UV auto zero steps in System performance method?

Repeat UV Auto zero. The back pressure should be 0.06 to 0.2 MPa at 5 ml/min.

What are the gradient steps in System performance method?

Start gradient, 0% to 100% B in 10 minutes, and start fractionation/collection.

What should you do after running the System performance method?

Make sure to update the Performance result text file with manually observed recordings and then print the report.

What is the purpose of switch valve timing optimization?

Switch valve timing optimization is used to optimize the switch valve (Buffer valve) timing of the ÄKTA start. It is recommended to optimize the timing of the switch valve when wavy gradients are obtained or when fluctuations in the step gradient are observed during either the System performance method or chromatography runs.

What are the default switch valve timings?

The default Switch valve timing A is 4 s, and Switch valve timing B is 5 s.

What is the adjustable range for switch valve timing?

The switch valve time can be set between 3.0 and 5.0 s with 0.1 s increments using Advanced timing.

How do you evaluate gradient fluctuations after changing switch valve timing?

After changing the Switch valve timing, perform a System performance method to evaluate the gradient fluctuations/wavy gradients, or a manual run of 50% B for 10 min can also be performed.

How do you set the switch valve timing?

In the Settings and service screen, tap System. In the System screen, tap Switch valve timing. Select the required timing by tapping the radio button for either Switch valve timing A (4 s) or Switch valve timing B (5 s). Then, tap Save to save the timing. If wavy gradients are still obtained, or if fluctuations on step gradient levels are large, then select Advanced timing. Set the switch valve time in the range of 3.0 to 5.0 s (0.1 s increments) by pressing the up/down arrows and tap Save.

Where is the column connected in the flow path?

The column is connected in the flow path between the Injection valve and UV Monitor.

Where are the column holder rails located on the instrument?

Column holder rails are located on the front and on the right side of the instrument.

How do you connect a column to the instrument?

Attach an appropriate column holder to the column holder rail on the instrument. Remove the column stoppers, and mount the column on the union connector if required. Fix the column to the column holder. Connect the 0.75 mm i.d. PEEK tubing from the Injection valve port 1 to the column head. Connect the 0.75 mm i.d. PEEK tubing from the bottom of the UV Monitor to the bottom of the column. Do not overtighten when connecting columns.

What is the purpose of the Pump wash A method?

The Pump wash A method is used before the start of a new run or when the buffers are changed. During Pump wash A, the flow is directed through the Wash valve to Waste.

How is Pump wash A performed?

Pump wash A is performed at 10 ml/min for 1 min through Buffer A port. It is recommended to perform the pump wash twice, first with DM water and then with the buffer of choice.

How do you initiate Pump wash A from the Instrument Display?

In the ÄKTA start home screen, tap Method run. In the Method run screen, tap Prepare system. Select Pump wash A and then tap Select to initiate the method. If required, the Pump wash A run can be ended before it is completed by tapping End to stop the run. When the run is completed, tap Exit to close the Pump wash A screen.

What is the purpose of the Pump wash B method?

The Pump wash B method is used before the start of a new run or when the buffers are changed. During Pump wash B the flow is directed through Wash valve to Waste.

How is Pump wash B performed?

Pump wash B is performed at 10 ml/min for 1 min through Buffer B port. It is recommended to perform the pump wash twice, first with DM water and then with the buffer of choice.

How do you initiate Pump wash B from the Instrument Display?

In the ÄKTA start home screen, tap Method run. In the Method run screen, tap Prepare system. Select Pump wash B and then tap Select to initiate the method. If required, the Pump wash B run can be ended before it is completed by tapping End to stop the wash in advance. When the run is completed, tap Exit to close the Pump wash B screen.

What is the purpose of the Washout fractionation tubing method?

The Washout fractionation tubing method is used to rinse the fractionation tubing. It is recommended when fractions are collected using the Outlet valve without a Fraction collector, and between different runs using the Fraction collector.

How does the Washout fractionation tubing method work?

Flow is diverted from the fractionation tubing to the collection tube, through the Outlet valve.

How do you initiate the Washout fractionation tubing method?

Immerse the buffer inlet tubing in DM water or buffer. Remove the column from the flow path and re-connect the flow path. Place the fractionation tubing in the waste container. In the ÄKTA start home screen, tap Method run. In the Method run screen, tap Prepare system. Select Washout fractionation tubing and then tap Select to initiate the method. Set the required run parameters, such as flow rate, pressure limit, and run volume. Tap Run to initiate the method. If required, the Washout fractionation tubing run can be ended before it is completed by tapping End to stop the washout in advance. When the run is completed, tap Exit to close the Washout fractionation tubing screen.

What is the purpose of the Column preparation method?

The Column preparation method is used to prepare a new column or to equilibrate the column.

How do you initiate the Column preparation method?

Immerse the buffer inlet tubing in the intended buffer. Connect the column into the flow path. In the ÄKTA start home screen, tap Method run. In the Method run screen, tap Prepare system. Select Column preparation and then tap Select to initiate the method. Set the required run parameters and then tap Run to initiate the method. If required, the Column preparation run can be ended before it is completed by tapping End to stop the run. When the run is completed, tap Exit to close the Column preparation screen.

What are the different modes of sample application available for ÄKTA start?

The different modes of sample application are via a Sample loop (25 µl to 10 ml), a Superloop (10 to 150 ml), or via the Pump from the Sample valve port I (sample) for volumes larger than 5 ml.

How does the Injection valve work?

The Injection valve enables the application of a sample onto the column from a sample loop connected to the valve. The Injection valve position can be changed manually by turning the lever to the left (Load Sample position) or to the right (Inject to column position).

What is the function of the Injection valve in the Load Sample position?

In the Load Sample position, the valve directs the system flow path from port 6 to port 1, and directs the liquid manually injected through port 3 to the sample loop. It also directs the liquid from the sample loop through port 4 to the waste container.

What is the function of the Injection valve in the Inject to column position?

In the Inject to column position, the valve diverts the system flow path to the sample loop (6 to 5) and directs the liquid from the sample loop to the column (2 to 1).

How do you connect a sample loop?

Connect the sample loop between ports 2 and 5 of the Injection valve. Make sure that waste tubing is connected to port 4 of the Injection valve.

How do you connect a Superloop?

Attach an appropriate column holder to the column holder rail on the right side edge of the instrument. Make sure that the Superloop is filled with liquid according to the Superloop instructions. Attach the Superloop to the column holder. Connect the tubing from the bottom of the Superloop to port 2 of the Injection valve. Connect the tubing from the top of the Superloop to port 5 of the Injection valve. Make sure that port 4 of the Injection valve is connected to waste.

How do you prime the sample tubing using the Pump?

Connect 1 mm i.d. ETFE tubing to port I (Sample) of the Sample valve. Immerse the other end of the sample inlet tubing in the DM water/Buffer container. From the Instrument Display, select Manual Run. Set the Sample valve position to Sample. Tap Run to start the run. End the run manually once the priming with the required volume of DM water or buffer has completed.

How do you load the sample using the Pump?

Connect 1 mm i.d. ETFE tubing to port I (Sample) of the Sample valve. Immerse the other end of the sample inlet tubing in the sample container. From the Instrument Display, select Method Run. In the Run parameters screen, select the sample application via the Pump. Set the sample volume and other required parameters. Tap Run to start the run. Make sure that the Injection valve is manually set to the Load Sample position.

How do you prime the sample loop before injecting the sample?

Fill a syringe with DM water or buffer. Make sure that the Injection valve is set to the Load Sample position. Connect the syringe to the Injection valve port 3. Load the DM water or buffer into the sample loop. Repeat the steps using a total of at least 5 times the loop volume before loading the sample.

How do you load the sample into the sample loop?

Fill a syringe with sample. Connect the syringe to the Injection valve port 3. Make sure that the Injection valve is set to position Load Sample. Carefully load the sample into the sample loop. Overfill the loop to make sure it is completely filled. From the Instrument Display, select Method run, then choose Templates. In the Run parameters screen, select the sample application via the Loop and set all the required run parameters. When prompted on the Instrument Display, switch the Injection valve position to the Inject to column position and tap Continue. When prompted on the Instrument Display, switch the Injection valve position to the Load Sample position and tap Continue.

How do you load the sample from a Superloop?

Fill a large volume syringe with sample. Connect the syringe to the Injection valve port 3 and carefully inject the sample into the Superloop. Make sure that the Injection valve is set to the Load Sample position. From the Instrument Display, select Method run, then choose Quick start methods or Templates. In the Run parameters screen, select the sample application via the Loop and set all the required run parameters. When prompted on the Instrument Display, switch the Injection valve position to the Inject to column position and tap Continue. When prompted on the Instrument Display, switch the Injection valve position to the Load Sample position and tap Continue.

How do you prepare the Fraction collector?

Insert a sufficient number of collection tubes into the Bowl assembly. Connect a 0.75 mm i.d. PEEK tubing to the Outlet valve port III (Collection). Loosen the nut of the tubing holder and insert the outlet tubing into the tubing holder. Tighten the nut. Fit the tubing holder into the corresponding port on the Dispenser arm. Gently move the arm to the dispensing position.

How do you set the delay volume?

In the ÄKTA start home screen, tap Settings and service to access the instrument modules. In the Settings and service screen, tap System to access the system options. In the System screen, tap Delay volume setting to access the settings. Enter the internal diameter (ID) in the Tube ID field and the length of the tubing from the Outlet valve to the Fraction collector in the Tube length fields, and then tap Save. The total delay volume from UV to Fraction collector is displayed in the Total delay volume field.

How do you prepare the instrument for operation in a cold room?

Place ÄKTA start in the cold room. If a UNICORN start computer is connected to the instrument, leave the computer outside the cold room. Allow the instrument to stabilize at the temperature of the cold room. Tighten all connections and pump DM water through the system to check for leaks. Tighten any leaking connector.

How do you remove the instrument from the cold room?

Switch off the instrument and disconnect the power cable before moving the instrument out of the cold room. Loosen all connections to prevent them from sticking when the system returns to room temperature. Allow the instrument to stabilize at room temperature for at least a few hours. Tighten all connections and pump DM water through the system to check for leaks. Tighten any leaking connector.

What checks should you make before starting a run?

Check that the buffer inlet tubings A and B are immersed in the correct bottles, and that there is sufficient buffer available. Check that the outlet tubings leading to waste from the Wash valve, Injection valve, and the Outlet valve are placed in the waste container, and that there is sufficient room in the waste container. If the fraction collector is going to be used, check that it is prepared and filled with appropriate collection tubes, and that the Outlet valve collection PEEK tubing is connected to the Fraction collector. Make sure that the Fraction collector is enabled. Check that the correct column has been connected and equilibrated. Make sure that the sample is ready to be loaded, and that the pump tubing is placed properly over the pump head and the pump head is properly closed. Make sure that a USB memory stick is connected to the instrument to save the results. If using UNICORN start, check that the system connection is established.

What are the options for starting a run from the instrument display?

The options for starting a run from the instrument display are Manual run and Method run.

What procedures are required after a run?

Procedures such as cleaning the column and the system flow path can be performed manually or using methods available in the Prepare system menu.

What are the options in the ÄKTA start home screen?

The ÄKTA start home screen displays four different options: Method run, Manual run, Create method, and Settings and service.

What should you check to make sure the system is correctly prepared?

The system should be prepared according to Chapter 5 and the modules calibrated according to Section 5.3. A suitable column has been selected, a suitable sample application technique will be used. The buffer inlet tubing is immersed into correct buffer bottles, and the waste tubing is inserted into an appropriate waste container. No tubing is twisted or blocked and the flow path is free from leakage. The Fraction collector configuration is either enabled or disabled as required. If the Fraction collector is used, make sure to use tubes with the same tube size. The delay volume is set. A USB memory stick is connected to the instrument.

What are the fractionation options offered by ÄKTA start?

ÄKTA start offers fixed volume fractionation, peak fractionation (level based and slope based), single peak collection (level based), and collection of elution volume.

How is the delay volume handled with and without Frac30?

With Frac30, the delay volume is collected in the first tube (T1). Without Frac30, the delay volume is collected in the collection container along with the first fraction.

What is the UV to Outlet valve volume for ÄKTA start?

The UV to Outlet valve volume is 0.27 ml for all ÄKTA start instruments if the recommended tubing dimensions are used.

How do you start a manual run?

In the ÄKTA start home screen, tap Manual run to access the run parameters for a manual run.

How do I set the run parameters for a manual run?

Use the up/down arrows or the numpad to set the values for flow rate, pressure limit, and buffer B concentration. If you want to save the result, tick the checkbox Save Result to USB. You can edit the result file name by setting the digits in the range 00 to 99. Tap the forward arrows to access additional run parameters. Tap Run when all required parameters have been set. Make sure that the values for the Flow rate and Pressure limit are appropriate for the chosen column.

What happens if the pressure exceeds the set limit during a manual run?

If the pressure exceeds the set limit, the instrument will enter the Pause state.

How do I set the valve positions for a manual run?

Set the sample valve as Buffer or Sample so that the flow is delivered from either the buffer inlets or the sample inlet. Set the wash valve as Column or Waste to direct the flow either to the column or to waste. Set the outlet valve as Collection or Waste to direct the flow either to the Fraction collector or to waste. Set the Fractionation volume, the volume of the fraction to be collected when the Fraction collector is enabled. Use up/down arrows to set the value, or use the numpad to type in the value. To collect fractions, place the required number of tubes of adequate volume in the Bowl assembly and make sure that the Fraction collector is enabled. If enabled, the Fraction collector will home to position 1 at the beginning of every run.

How do I start a manual run?

Tap Run to start the run. The Run view screen will be displayed.

How do I monitor and control a manual run?

From the Run view screen, the user can monitor and control the ongoing run. The following options are available:

Graph: Displays the run-time UV absorbance curve.
Edit run: Allows the user to edit the run parameters of the ongoing run.
Pause: Temporarily pauses the run by stopping the pump.
End: Terminates the current run.

How do I view the chromatogram of an ongoing manual run?

In the Run view screen, tap the graph icon to view the chromatogram. The graph displays the UV curve. The Y axis displays the UV absorbance (mAU) and X axis the time (min). Tap the return arrow to return to the Run view screen.

How do I edit the run parameters of an ongoing manual run?

In the Run view screen, tap Edit run to access the run parameters of the ongoing run. Edit the run parameters:

Buffer B concentration (%)
Flow rate (ml/min)
Fractionation volume (ml)
Toggle Fractionation between Start and Stop to start or stop the fractionation.

Use up/down arrows to adjust the values, or use the numpad to type in the values. If no other parameters need to be set, tap Execute to implement the changes. To ignore the changes, tap Cancel. Tap the forward arrow to access additional run parameters. Toggle as needed to set which valve positions are open:

Sample valve: set as Buffer or Sample so that the flow is delivered from either the buffer inlets or the sample inlet
Wash valve: set as Column or Waste to direct the flow to either the column or to waste
Outlet valve: set as Collection or Waste to direct the flow to either the Fraction collector or to waste

Tick the Autozero UV checkbox if a UV baseline to zero is required. Tick the Set event mark checkbox if you need to set an event mark in the chromatogram. After you have set the run parameters, tap Execute to implement the changes.

How do I pause a manual run?

In the Run view screen, tap Pause to temporarily pause the run by stopping the pump. To continue the run, tap Continue. There is no liquid flow in the flow path when the run is paused.

How do I end a manual run?

In the Run view screen, tap End to terminate the run. A Message screen opens, requiring confirmation of the action. Tap Yes to confirm that you want to terminate the run or tap No to cancel the action and return to the Run view screen. When a run is terminated before it is completed, the partial result is stored on the USB memory stick. The USB memory stick stores the result files which can be viewed using UNICORN start. Also a BMP file is generated which can be viewed from any PC. Do not remove the USB memory stick before the save operation is complete. Tap Exit to close the Run ended screen.

What are the different method types available?

The different method types are:

Quick start: Allows the user to run methods like affinity, Ion exchange, Gel filtration and Desalting with method parameters predefined.
Templates: Allows the user to edit and run the predefined methods Affinity, Ion exchange, Gel filtration and Desalting.
User defined: Allows the user to run user created methods or USB imported methods.
Prepare system: Allows the user to perform system operations, such as Pump wash, Column preparation, Cleaning and System performance test.

What are the options under Quick Start?

The options under Quick Start are:

AC step 1 ml HiTrap
AC step 5 ml HiTrap
DS 5 ml HiTrap
DS 53 ml HiPrep
IEX step 1 ml HiTrap
IEX step 5 ml HiTrap
IEX gradient 1 ml HiTrap
IEX gradient 5 ml HiTrap
GF 16/60 HiPrep

What are the options under Templates?

The options under Templates are:

Affinity (AC)
Desalting/buffer exchange (DS)
Ion exchange (IEX)
Gel filtration (GF)

What are the options under Prepare System?

The options under Prepare System are:

Pump wash A
Pump wash B
Washout fractionation tubing
Column preparation
System cleaning
System performance method

How do I select a method?

In the ÄKTA start home screen, tap Method run to access the method types available with the instrument. Select one of the following methods:

Quick start
Templates
User defined
Prepare system

What is Quick Start?

Quick start contains “ready to run” methods to purify most common proteins based on Affinity, Ion exchange, Gel filtration and Desalting techniques. Run parameters like column volume, flow rate, equilibration and elution mode, and volume are predefined in the method. The user needs to enter only the sample volume. If required, the run parameters can be changed using the Edit run option during an ongoing run.

What are the different Quick Start techniques?

Affinity Chromatography (AC step 1 ml/5 ml HiTrap): Bound proteins are eluted in a single step, using a single elution buffer. Commonly used for purification of tagged proteins, for example Histidine-tagged proteins.
Desalting (DS 5 ml HiTrap/53 ml HiPrep): Proteins are eluted in a single step, using a single elution buffer.
Ion Exchange Chromatography (IEX step 1 ml/5 ml HiTrap): Bound proteins are eluted in a single, step using a single elution buffer.
Ion Exchange Chromatography (IEX gradient 1 ml/5 ml HiTrap): Bound proteins are eluted using two buffers with linear increase in the concen-tration of buffer B, over a specified time followed by a step of 100% B.
Gel Filtration (GF 16/60 HiPrep): Proteins are eluted in a single step, using a single elution buffer. It is recommended to use the appropriate column as indicated in the template names.

How do I start a run using a Quick Start method?

Make sure that a USB memory stick is connected to the instrument. In the Method run screen, tap Quick start to access the templates. To select a Quick start method, tap a radio button or tap the forward arrow to access additional Quick start methods. To continue with the selected method, tap Select. Make sure to load the recommended sample volume for the selected column. If large Gel filtration columns are used, it is recommended to pre-equilibrate the column before starting the run. The mode of sample application is Pump (default). Sample application using Pump is used for all Quick start methods to automate sample loading or to have an unattended chromatographic run. Sample application via Loop is not applicable. Enter the sample volume in the Sample volume field. Use up/down arrows to set the values, or use numpad to type in the values. Tick the checkbox Save Result to USB if you want to save the result. The result file name can be edited by setting the digits in the range 00 to 99. Tap Run to start the run. The Run view will be displayed. Other run parameters can be edited using the Edit run option available in the Run view screen. In the Run view screen, the following options are available to monitor and control the ongoing run:

Graph, to view the chromatogram.
Edit run, to change any run parameters in the current run.
Hold, to temporarily hold the run, with current set flow rate, valve positions and B concentration.
Pause, to pause the current run.
End, to end the run before it is completed. The run begins with a default pump wash. Pump wash is performed at 10 ml/min for 1 min with 30 s of buffer B wash followed by 30 s of buffer A wash. Edit run is disabled when the pump wash is in progress. During pump wash the flow is directed through Wash valve to Waste. The USB memory stick stores the result files which can be viewed using UNICORN start. Also a BMP file is generated which can be viewed from any PC. Do not remove the USB memory stick until the system generates the report.

How do I hold a Quick Start run?

In the Run view, tap Hold to temporarily hold the run. The Hold option is active only in a method run and is not applicable for a manual run. To resume the run, tap Resume. During hold the run is temporarily interrupted. Flow continues at the current flow rate, but the current gradient concentration and valve positions are maintained.

What are the predefined method templates?

The predefined templates are:

Affinity (AC): Affinity Chromatography separates molecules based on the reversible interaction between the target protein and the specific ligand attached to a chromatography matrix.
Ion exchange (IEX): Ion Exchange Chromatography is based on the reversible interaction between a charged protein and an oppositely charged chromatography medium.
Gel filtration (GF): Gel filtration, also known as size-exclusion chromatography, is a chromatography technique that separates molecules based on differences in the molecular size.
Desalting/buffer exchange (DS): Desalting is a gel filtration technique that allows rapid group separation of high molecular weight substances from low molecular weight substances. Small molecules like salt, free labels and other impurities are efficiently separated from the high molecular weight substances of interest.

How do I start a run based on Affinity (AC) or Ion exchange (IEX) Templates?

Make sure that a USB memory stick is connected to the instrument. Before starting the run, set the status of the Fraction collector to enabled or disabled as required. In the Method run screen, tap Templates to access the different templates. Tap a radio button to select a template that suits your application. To continue with the selected technique, tap Select. Set the run parameters:

Column volume (ml)
Flow rate (ml/min)
Pressure limit (MPa)

Tick the checkbox Save Result to USB if you want to save the result. The result file name can be edited by setting the digits in the range 00 to 99. Tap the forward arrow to access additional run parameters. Make sure that the values for the Column volume, Flow rate and Pressure limit are appropriate for the chosen column. If the pressure reaches above the set limit, the instrument will enter the Pause state. The USB memory stick stores the result files which can be viewed using UNICORN start. Also a BMP file is generated which can be viewed from any PC. Do not remove the USB memory stick until the system generates the report (BMP file). Select the mode of sample injection in the Sample from field: to be applied via Pump or via Loop. Set the run parameters:

Sample volume, the volume of sample to be loaded onto the column
Equilibration volume, the volume of buffer A required for for equilibrating the column
Wash unbound volume, volume of buffer needed after the sample application to wash off the unbound molecules For AC/IEX methods when loading sample through loop it is advisable to empty the loop with 3 times the loop volume to achieve good sample recovery. Configure the run parameters for Elution Option set as Isocratic:
Concentration of buffer B to elute the bound protein
Elution volume, volume needed to elute the bound protein from the column
Fractionation volume, volume of the fraction to be collected when the Fraction collector is enabled or Configure the run parameters for Elution Option set as Gradient:
Target conc B, maximum buffer B concentration level to be set in the gradient
Gradient volume, volume needed to elute the bound protein from the column
Fractionation volume, volume of the fraction to be collected when the Fraction collector is enabled.

Tap back arrow to view or edit run parameters. Tap Run to start the run. The Run view screen will be displayed. The run begins with a default pump wash. Pump wash is performed at 10 ml/min for 1 min with 30 s of buffer B wash followed by 30 s of buffer A wash. Edit run is disabled when the pump wash is in progress. During pump wash the flow is directed through Wash valve to Waste.

How do I start a run based on Gel Filtration or Desalting/Buffer Exchange Templates?

Make sure that a USB memory stick is connected to the instrument. Before starting the run, set the status of the Fraction collector to enabled or disabled as required. In the Method run screen, tap Templates to access the predefined method templates. Tap a radio button to select a template that suits your application (e.g., Gel filtration). To continue with the selected technique, tap Select. Set the run parameters:

Column volume (ml)
Flow rate (ml/min)
Pressure limit (MPa) Use up/down arrows to set the values, or use numpad to type in the values. Tick the checkbox Save Result to USB if you want to save the result. The result file name can be edited by setting the digits in the range 00 to 99. Tap the forward arrow to access additional run parameters. Make sure that the values for the Flow rate and Pressure limit are appropriate for the chosen column. If the pressure reaches above the set limit, the instrument will enter the Pause state. The USB memory stick stores the result files which can be viewed using UNICORN start. Also a BMP file is generated which can be viewed from any PC. Do not remove the USB memory stick until the system generates the report (BMP file). Select the mode of sample injection in the Sample from field: to be applied via Pump or via Loop. Set the run parameters:
Sample volume, the volume of sample to be loaded in to the column
Equilibration volume, the volume of buffer A required for for equilibrating the column. Washout unbound is not applicable for GF/DS methods. Make sure to load the recommended sample volume for selected column. If large GF columns are used, it is recommended to pre-equilibrate the column before starting the run. Set the run parameters:
Elution volume, volume of buffer needed to elute the protein from the column
Fractionation volume, volume of fractions to be collected when the Fraction collector is enabled. Tap Run to start the run. The Run view screen will be displayed. Conc B is not applicable for GF/DS as elution takes place only with a single buffer (buffer A). The run begins with a default pump wash. Pump wash is performed with buffer A at 10 ml/min for 30 s. Edit run is disabled when the pump wash is in progress. During pump wash the flow is directed through Wash valve to Waste.

How do I start a run based on a user created or USB imported method?

Make sure that a USB memory stick is connected to the instrument. In the Method run screen, tap User defined to access the user created methods. Tap a radio button to select a user method to run. To continue with the selected user method, tap Select. Set the run parameters:

Select the mode of sample injection in the Sample from field: to be applied via Pump or via Loop
Sample volume, the volume of sample to be loaded in to the column
Tick the checkbox Save Result to USB if you want to save the result. The result file name can be edited by setting the digits in the range 00 to 99. Tap Run to start the selected method. The Run view screen will be displayed. In the Run view screen, the following options are available to monitor and control the ongoing run:
Graph, to view the chromatogram.
Edit run, to change any run parameters in the current run.
Hold, to hold the current run.
Pause, to pause the current run.
End, to end the run before it is completed. The USB memory stick stores the result files which can be viewed using UNICORN start. Also a bmp file is generated which can be viewed from any PC. Do not remove the USB memory stick until the system generates the report (.bmp file). USB imported methods which were created in UNICORN start cannot be edited in the instrument. Use UNICORN start to edit those methods.

What are the Prepare System methods?

The Prepare System methods are:

Pump wash A
Pump wash B
Washout fractionation tubing
Column preparation
System cleaning
System performance method

The Pump wash A/B, Washout fractionation tubing, and Column preparation methods are required for the system preparation. The System performance method is also available.

How do I perform a system cleaning run?

In the Method run screen, tap Prepare system to access the system methods. In the Select system method screen, select System cleaning. To continue with the System cleaning method, tap Select. Perform the operations described on the display:

Disconnect the column.
Connect Injection valve to UV with suitable tubing and connectors.
Place the buffer tubings (A & B) in cleaning solution. Tap Continue to start the System cleaning run. Wait for the run to complete. If required, the System cleaning run can be ended before it completes by tapping End. Tap Exit to close the System cleaning screen when the System cleaning run is completed.

How do I evaluate a run?

After a chromatographic run, the result stored on a USB memory stick can be transferred to UNICORN start where it can be viewed, and evaluated. The result holds a complete record of the run, including method, system settings, chromatogram, and run log. The result can be also viewed by using the BMP result file that is generated and stored on the USB memory stick.

How do I clean the system after a run?

After a run is completed, perform the following:

Remove the column from the flow path and re-connect the flow path.
Rinse the flow path with cleaning solution and/or DM water using either System cleaning or the Pump wash methods, as required.
If required, remove the tubes from the Fraction collector. If there is any spillage, clean the Bowl assembly with DM water.
Clean all spills on the instrument and on the bench using a damp cloth.
Empty the waste container.

How do I clean and store the column after a run?

After a run is completed, perform the following:

Disconnect the column from the flow path.
Clean the column off-line according to the column instructions. If the column is not going to be used for a couple of days or longer, perform the following:
Fill the column with the storage solution recommended in the column data sheet.
Detach the column from the instrument and store it according to the column recommendation.

How do I store the system for a period of time?

If the instrument is not going to be used for a period of time, fill the system and the inlets with storage solution (DM water or 20% ethanol).

How do I power off the instrument?

Switch off the instrument by turning the Power switch to the O position. When the instrument is switched off or not in use, make sure to open the pump hood and release the pump tubing from the pump head.

What does the Create method menu allow?

The Create method menu allows the user to create a new method, edit, import, and delete a method from the ÄKTA start Instrument Display.

How do I access the Create method options?

In the ÄKTA start home screen, tap Create method. In the Create method screen, the following options are available:

Create, to create a new method using a predefined template
Edit, to edit a method or change run parameters for user created methods stored on instrument
USB Import, to import a method developed on UNICORN start to the instrument using a USB memory stick
Delete, to delete a method that is stored on the instrument

How do I create a method using a predefined template?

In the Create method screen, tap Create. The Select templates screen opens. Tap a radio button to select a template. Tap Create to create a method based on the selected technique. Set the run parameters:

Column volume (ml)
Flow rate (ml/min)
Pressure limit (MPa) Select Save Method as field if you want to set a method name. The file name can be edited by setting the digits in the range 00 to 99. Provide a unique method name. The created method will be saved under the User defined methods menu. Make sure that the values for the Column volume, Flow rate and Pressure limit are appropriate for the chosen column. If the pressure reaches above the set limit, the instrument will enter the Pause state. Select the mode of sample injection in the Sample from field: to be applied via Pump or via Loop. Set the run parameters:
Sample volume, the volume of sample to be loaded onto the column
Equilibration volume, the volume of buffer A required for for equilibrating the column
Wash unbound volume, volume of buffer needed after the sample application to wash off the unbound molecules. The Washout unbound volume is applicable only for AC/IEX techniques. Configure the run parameters for Elution Option set as Isocratic:
Concentration of buffer B to elute the bound protein
Elution volume, volume needed to elute the bound protein from the column
Fractionation volume, volume of the fraction to be collected when the Fraction collector is enabled or Configure the run parameters for Elution Option set as Gradient:
Target conc B, maximum buffer B concentration level to be set in the gradient
Gradient volume, volume needed to elute the bound protein from the column
Fractionation volume, volume of the fraction to be collected when the Fraction collector is enabled. Tap Save to save the new method. A Message screen that requires to confirm the action will be displayed. Tap Yes to confirm the saving of the method or tap No to cancel the action and return to set the run parameters. The created method will be saved under the User defined methods menu. The system can store up to 10 methods only. Delete existing methods if new methods need to be stored.

How do I edit a user defined method?

In the Create method screen, tap Edit to access the methods. The Select method to edit screen opens. In the Select method to edit screen, tap a radio button to select a user method. Tap Edit to start editing the run parameters for the selected method. Select Save Method as field if you want to set a method name. The file name can be edited by setting the digits in the range 00 to 99. Provide a unique method name. The created method will be saved under the User defined methods menu. USB imported methods which were created using UNICORN start cannot be edited from the instrument. Use UNICORN start to edit those methods. Tap Yes to confirm the saving of the method or tap No to cancel the action and return to set the run parameters.

How do I import a method stored on a USB memory stick?

Make sure that the USB memory stick containing the user defined methods is connected to the instrument. In the Create method screen, tap USB Import to access the methods. Tap a radio button to select a method. Tap Import to import the method. A Message screen that requires to confirm the action will be displayed. Tap Yes to confirm the import of the selected file or tap No to cancel the action and return to the file list. The imported methods will be saved under the User defined methods menu. Only one method can be imported at a time. If multiple methods are to be imported, repeat the steps described above. If the system memory is full, delete existing methods before importing a new method.

How do I delete a user method?

In the Create method screen, tap Delete to access the methods. Tap a checkbox to select a method. Tap Delete to delete the method. A Message screen that requires to confirm the action opens. To cancel the action and return to the Create method screen, tap Cancel. Multiple files can be deleted at the same time. Tap Yes to confirm the deletion of the selected files or tap No to cancel the action and return to the file list.

What is a USB memory stick used for?

The USB memory stick is used to save a result, BMP file and also for transferring methods between the instrument and UNICORN start. The USB memory stick is also used to generate a System error report. The result files will be saved in the GE folder which is automatically created by the instrument when the USB memory stick is first plugged in. At any given point of time only 10 results can be stored in the GE folder. To save further results, transfer the result files to another folder, PC or re-name the GE folder.

How do I store results on a USB memory stick?

Connect a USB memory stick to the instrument, via the USB port. Start a Manual run or Method run. In the Run parameters screen, tick the Save results to USB check box in order to save the generated results on the USB memory stick. When the run is complete, the results are saved in a GE folder. A BMP result file is also generated. Hence, the chromatogram can be viewed without the use of UNICORN start. Export the results to UNICORN start to view the chromatogram and evaluate. Make sure that the USB memory stick is not removed during the run.

What should I do and not do while handling the USB memory stick?

Make sure that the USB memory stick is inserted completely into the instrument.
The maximum supported USB memory stick capacity is 32 GB. A minimum of 1 GB free space is required to execute read/write operations.
Only FAT32 file system is supported and this needs to be taken into account when formatting the memory stick.
Only unplug the USB memory stick when the instrument display is in the Home screen.
It is preferred to keep a minimum number of files on the memory stick. Once you take a backup of the files, delete them from the memory stick and then save onto a computer. Avoid keeping unnecessary files in the memory stick.
Avoid using folders named GE on the USB memory stick. However, you may use folders named for example GE_ or GExyz.
Always take backup of complete GE folder from USB memory stick and do not backup individual files. It is recommended to take backups whenever you have completed important runs.

How do I import a result file from a USB stick to UNICORN start?

Open the Evaluation module in UNICORN start. Select File:Import:Import ÄKTA start results from USB…, and then import the result files to a desired location on the computer. View, analyze, report or print the result file.

How do I export a method from UNICORN start to a USB memory stick?

Create a method using the Method Editor module in UNICORN start. Connect a USB memory stick to the computer. Select File:Export:Export Method… in order to export the created method to a USB memory stick, connected to the computer. Make sure that the method is stored in a GE folder.

How do I import a method to ÄKTA start from UNICORN start using a USB stick?

Create a method using the Method Editor module in UNICORN start. Select File:Export:Export Method for ÄKTA start to USB… in order to export the created method to a USB memory stick. Connect the USB memory stick to ÄKTA start. Make sure that the method is stored in a GE folder. From the ÄKTA start Home screen, tap Create method : USB import. Select the method to import.

What is the BMP result file?

The BMP result file allows the user to view the result image using another type of software outside of ÄKTA start and UNICORN start. This format facilitates viewing of the generated result without UNICORN start.

What are the features of the exported BMP result file?

The result file is a graphics file in BMP format, compatible with Microsoft® Windows® and Macintosh™ operating systems.
The BMP result is saved and exported, if the user selects the option to Save result to USB before the start of a run.
The result file contains the UV curve data with fractionation marks.
The result file contains up to 4 hours of run data. For longer runs, the last 4 hours are saved.
The BMP file displays the necessary legends like Product name, run details, UV, Frac marks.
The result file is saved if the user terminates the run before completion. However, no file is saved in the event of shutdown or power failure.

What is the main function of the Method Editor module?

The Method Editor module provides flexibility to create or edit the chromatography methods.

What are the main features of the Method Editor module?

Ability to create methods from predefined templates like Affinity, Ion Exchange, Desalting and Gel Filtration.
Flexibility to create customized methods by dragging and dropping chromatography phases such as Prime and Equilibration, Sample Application, Wash Out Unbound, Elution and Fractionation.

What can be done with the methods created from Method Editor?

The methods created from Method Editor can either be run directly from System Control or exported to a USB memory stick to run them from ÄKTA start directly.

What is the main function of the Evaluation module?

The Evaluation module is used to manage and evaluate the results from chromatography runs.

What are the main features of the Evaluation module?

Open and view existing chromatogram results.
Compare two curves or chromatograms.
Perform peak integration analysis.
Create and print PDF reports.
Import results from ÄKTA start via a USB memory stick.

What is the main function of the Administration module?

The Administration module is used to manage the UNICORN start database and to review UNICORN start and system logs.

What are the main features of the Administration module?

Ability to back up/restore and archive/retrieve files.
Review the UNICORN start and system logs.

What regular maintenance should be performed on the system?

Regular maintenance should be performed on a daily, weekly and monthly basis.

What daily maintenance operations should be performed when the system is in use?

Visually inspect the instrument for leakages in the flow path.
Check the Pump for leakage. If there are signs of liquid leaking from the Pump, check the integrity of the pump tubing and the tubing connections.
Make sure that the pump tubing is not left in-side the Pump when it is not in use.
Clean the column and the system flow path after use and leave the system filled with DM water.

What should you do if the instrument is not going to be used for a few days?

If the instrument is not going to be used for a few days, prepare the system for storage.

What should be done with the valves after every run, or at the end of the day, or while leaving the instrument idle for several days?

Clean the valves (at least with DM water), after every run, or at the end of the day, or while leaving the instrument idle for several days to avoid salt crystal formation.

What can be used if a more thorough cleaning is required for the valves?

If more thorough cleaning is required, 1 M NaOH may be used. After using NaOH for cleaning make sure to wash the flow path thoroughly with DM water before starting a run.

What weekly maintenance operations should be performed?

Calibrate the Pump.
Visually inspect the inlet filters and clean them if necessary.

What monthly maintenance operations should be performed?

Clean the system flow path with 1 M NaOH and rinse with DM water.
Visually inspect the drive sleeve on the Fraction collector. Replace if worn out.

What other maintenance operations should be performed when required?

Clean the instrument externally.
Clean the Fraction collector.
Perform System cleaning.
Clean the UV flow cell.
Clean the Conductivity cell.
Calibrate the touch screen.
Calibrate the UV flow cell.
Calibrate the Conductivity cell.
Pressure sensor zero offset.
Replace the inlet filters.
Replace the tubing and connectors.

What should be done before planned maintenance or service?

To ensure the protection and safety of service personnel, all equipment and work areas must be clean and free of any hazardous contaminants before a Service Engineer starts maintenance work.

What should be completed before planned maintenance or service?

Please complete the checklist in the On Site Service Health & Safety Declaration Form, or the Health & Safety Declaration Form for Product Return or Servicing, depending on whether the instrument is going to be serviced on site or returned for service.

Why is cleaning the system flow path performed?

Cleaning the system flow path is performed to prevent carryover between runs, contamination in the flow path and as a routine maintenance protocol.

How is cleaning the system flow path usually performed?

Cleaning the system flow path is usually performed by using System cleaning or Pump wash methods.

What should be done before cleaning the system flow path?

Before cleaning the system flow path, remove the column from the flow path.

What should be done when using hazardous biological agents during a run?

When using hazardous biological agents, run the System cleaning method to flush the entire system tubing with 1 M NaOH and subsequently with distilled water, before service and maintenance.

What precautions should be taken when using NaOH?

NaOH is corrosive and therefore dangerous to health. When using hazardous chemicals, avoid spillage and wear protective glasses and other suitable Personal Protective Equipment (PPE).

What protective measures should be taken when using hazardous chemical and biological agents?

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.

What should be done if hazardous chemicals are used for system or column cleaning?

If hazardous chemicals are used for system or column cleaning, wash the system or column with a non-hazardous solution in the last phase or step.

What should be done before cleaning the system flow path?

The column should be removed from the flow path before cleaning the system flow path.

How should the flow path be re-connected after the column is removed?

The flow path has to be re-connected between the manual Injection valve port 1 and the UV inlet.

What should be done for column cleaning procedures and storage instructions?

For column cleaning procedures and storage instructions, refer to the column catalogue.

How should you remove the column and re-connect the flow path?

Disconnect the tubing from the column.
Re-connect the flow path between the Injection valve and the UV Monitor, using the union mounted on the tubing connected to the UV flow cell.

Why is the System cleaning method used?

The System cleaning method is used to clean the instrument flow path. System cleaning is recommended to be performed to prevent carryover between runs, contamination in the flow path, as a routine maintenance protocol and to prepare system for storage.

Why is cleaning important?

Cleaning is important for preventing cross-contamination and bacterial growth in the instrument.

What should you do to ensure proper cleaning?

Prepare cleaning solutions of recommended concentration to ensure proper cleaning.

Is it recommended to end the run before completion?

It is recommended not to end the run before completion.

What inlets and outlets should be cleaned from the Edit run screen?

It is recommended to clean the inlets and outlets (sample tubing, fraction-ation tubing) from the Edit run screen.

What solutions are required for System cleaning?

1 M NaOH
DM water

How should you clean the system flow path?

Remove the column from the flow path and re-connect the tubing.
Immerse both the buffer inlets in 1 M NaOH.
In the ÄKTA start home screen, tap Method run.
In the Method run screen, tap Prepare system.
Select System cleaning and then tap Select to initiate the method.
Perform the operations presented on the display:
- Disconnect the column if this has not already been done.
- Connect Injection valve to UV with suitable tubing and connectors.
- Place the buffer tubings (A & B) in cleaning solution.
Tap Continue to start System cleaning.
When the run is completed, tap End to close the System cleaning screen.
Wash the complete flow path with water to remove NaOH from the system. Check the pH after washing with water to make sure that all NaOH is re-moved.

How often should the UV flow cell be cleaned?

Clean the UV flow cell every six months, or when required.

Why is a clean flow cell essential?

A clean flow cell is essential for correct performance of the UV Monitor.

What should you not allow in the UV flow cell?

Do not allow solutions containing dissolved salts, proteins or other solutes to dry out in the UV flow cell. Do not allow particles to enter the UV flow cell, as damage to the flow cell may occur.

What solutions are required for cleaning the UV flow cell?

Cleaning solution: 10% detergent solution, such as Decon™ 90, Deconex™ 11, RBS 25, 1 M HCl or 1 M NaOH.
DM water

What is the recommended solution for cleaning the UV flow cell?

It is recommended to use 10% detergent solution for cleaning the UV flow cell.

How can you increase the cleaning efficiency of the 10% detergent solution?

Heat the 10% detergent solution to 40°C to increase the cleaning efficiency.

What should you do if NaOH is used for cleaning the UV flow cell?

If NaOH is used, perform cleaning at 1 ml/min and reduce the hold time to 5 min in step 3 of the method described below.

How long should NaOH be left in the flow cell and what should be done after?

NaOH should not be left in the flow cell for more than 20 minutes and proper care should be taken to remove the NaOH completely from the flow cell.

How should the UV flow cell be cleaned in-place?

Before cleaning the UV flow cell, remove the column from the flow path and re-connect the flow path.
Immerse the inlet tubing in cleaning solution.
Start a manual run and pump cleaning solution at 5 ml/min through the UV flow cell for 10 minutes and pause the run.
Leave the UV flow cell filled with cleaning solution for 15 minutes.
Immerse the inlet tubing in DM water.
Resume the run and rinse the flow cell thoroughly with DM water.

When should the Conductivity flow cell be cleaned?

Clean the Conductivity flow cell when the Conductivity Monitor shows a slow response or when the conductivity measurements are not comparable to previous results.

What solutions are required for cleaning the Conductivity flow cell?

1 M NaOH
DM water

How should you clean the Conductivity flow cell in-place?

Before cleaning the Conductivity flow cell, remove the column from the flow path and re-connect the flow path.
Immerse the inlet tubing (either A or B) in 1 M NaOH.
Start a manual run and pump 1 M NaOH at 1 ml/min through the flow cell for 10 minutes.
Pause the run. Leave the Conductivity flow cell filled with 1 M NaOH for 15 minutes.
Immerse the inlet tubing in DM water.
Resume the run and rinse the flow cell thoroughly.

How long should NaOH be left in the Conductivity flow cell and what should be done after?

Do not leave NaOH in the flow cell for more than 20 min to avoid damage. Rinse the flow path with water thoroughly.

How can you ensure that the NaOH is completely removed from the Conductivity flow cell?

Make sure that the NaOH is completely removed. The conductivity reading in the Run view screen should be < 1 mS/cm.

Why is valve cleaning required?

Valve cleaning is required to avoid any cross-contamination or salt crystal formation. Salt deposits may form if the flow path is not flushed out to remove buffer solutions after performing a run.

When is it recommended to clean the valves?

It is recommended to clean the valves (at least with DM water) after every run, or at the end of the day, or while leaving the instrument idle for several days.

What are the different protocols for cleaning the valves?

Cleaning the Buffer valve and Wash valve
Cleaning the Sample valve using ÄKTA start stand-alone
Cleaning the Sample valve using UNICORN start

How can the Buffer valve and Wash valve be cleaned?

The Buffer valve and Wash valve can be cleaned using DM water.

What can be used if a more thorough cleaning is required for the Buffer valve and Wash valve?

If more thorough cleaning is required, 1 M NaOH may be used. When NaOH is used for cleaning, make sure to wash the flow path thoroughly with DM water before starting a run.

How should the Buffer valve and Wash valve be cleaned?

Perform Pump wash B and Pump wash A for cleaning the buffer valve and wash valve (Pump wash flow rate is 10 ml/min).
If using NaOH for cleaning, repeat the Pump wash B and Pump wash A protocols with DM water until the NaOH is completely removed.

How can the Sample valve be cleaned using ÄKTA start stand-alone?

The Sample valve can be cleaned using DM water.

What can be used if a more thorough cleaning is required for the Sample valve?

If a more thorough cleaning is required, 1 M NaOH may be used. When NaOH is used for cleaning, make sure the flow path is thoroughly washed with DM water before starting a run.

How should the Sample valve be cleaned using ÄKTA start stand-alone?

Immerse the sample inlet tubing in the cleaning solution (DM water or 1 M NaOH).
In the ÄKTA start home screen, tap Manual run.
In the Manual run screen (1/2), set the Flow rate to 5 ml/min. Clear Save Result to USB. Tap the forward arrow to go to Manual run screen 2/2.
Set Sample valve to Sample. Set Wash valve to Column. Set Outlet valve to Waste.
Tap Run to start the run.
After 3 to 5 minutes, tap End to end the run.

How should the Sample valve be cleaned using UNICORN start?

Immerse the sample inlet tubing into the cleaning solution (DM water or 1 M NaOH).
Start the computer and launch UNICORN start and connect to ÄKTA start.
In the UNICORN start System Control module, click Manual run.
In the Manual Run settings dialog, set the Flow Rate to 5 ml/min and click OK.
In the process picture, set Sample valve to Sample and Wash valve to Column position.
After 3 to 5 minutes, end the run.

When should the inlet filters be cleaned?

Clean the inlet filters when required, for example when the visual inspection shows that the filters are clogged.

What solutions are required for cleaning the inlet filters?

1 M NaOH
DM water

How should the inlet filters be cleaned?

Pull off the support net and the inlet filter from the inlet filter holder.
Immerse and leave the inlet filter and the support net in 1 M NaOH for about 2 hours. Alternatively use a shorter time in an ultrasonic bath.
Remove the inlet filter and the support net from the NaOH solution and rinse thoroughly with DM water.
Fit the inlet filter into the support net, and press it into position on the inlet filter holder.

When should the instrument be cleaned externally?

Clean the instrument externally when required. Do not allow spilled liquid to dry on the instrument.

What materials are required for cleaning the instrument externally?

Cleaning cloth
Mild cleaning agent or 20% ethanol

How should the instrument be cleaned externally?

Check that no run is in progress.
Switch off the instrument.
Wipe the surface with a damp cloth. Wipe off stains using a mild cleaning agent or 20% ethanol.
Let the instrument dry completely before using it.

When should the fraction collector be cleaned?

Clean the fraction collector when required, for example in case of liquid spill in the Bowl assembly.

What materials are required for cleaning the Bowl assembly?

Water
20% ethanol
Cleaning cloth

How should the Bowl assembly be disassembled and cleaned?

Check that no run is in progress.
Switch off ÄKTA start and disconnect the Frac30 Cable.
Remove the collection tubes and disassemble the Bowl assembly from the Base unit:
- Gently move the Dispenser arm counterclockwise to the end position.
- Push the drive assembly and hold it at the retracted position.
- Lift and remove the Bowl assembly.
Wash the Bowl under a water tap. Use a mild detergent if required, and rinse thoroughly with water.
Wipe the surface with a damp cloth. Wipe off stains using water.
Let the Bowl assembly dry completely before re-assembling.
Re-assemble the Bowl assembly on to the Base unit:
- Orient the Bowl to match the aligning groove and the aligning features on the Bowl holder.
- Lower the bowl assembly on to the Base unit and push the drive as-sembly to allow the Bowl assembly to get in position.

When should the tube holder be removed from the bowl?

Do not remove the tube holder from the bowl except when cleaning is necessary. Frequent removal may damage the snap locks.

How do you remove the tube holder in the Bowl assembly?

To remove the tube holder, unsnap the snap locks one by one and sequentially for ease of removal.

How do you refit the tube holder in the Bowl assembly?

Align the holder with the help of the aligning groove.
Snap fit the part by operating one snap at a time in a continuous se-quence by pressing on the edge of the tube holder.

What materials are required for storing the instrument?

DM water
20% ethanol
0.75 mm i.d. PEEK tubing
Waste container

How do you prepare the system for short term storage?

Before cleaning the flow path, remove the column and re-connect the flow path.
Immerse the buffer and the sample inlet tubing in DM water.
In the ÄKTA start home screen, tap Manual run.
Set the run parameters according to the table:
- Flow rate: 1 ml/min
- Pressure limit: 0.5 MPa
- Buffer: Select Buffer/Sample
- Column: Wash valve
- Waste: Outlet valve
Tap Run to start the manual run.
Pump 20 ml of DM water through the system.
Tap Edit run and set the run parameters as indicated below:
- Sample: Select Buffer/Sample
- Collection: Outlet valve
Pump 20 ml of DM water through the system.
End the run and leave the system filled with DM water during the storage period.

How do you prepare the system for long term storage?

Before cleaning the flow path, remove the column and re-connect the flow path.
Immerse the buffer and the sample inlet tubing in 20% ethanol.
Start a manual run and pump 20 ml of 20% ethanol through the system. Use the same run parameters recommended for the short term storage procedure.
Edit the run and set the run parameters to clean the sample inlet tubing and the outlet tubing for fraction collection.
Pump 20 ml of 20% ethanol through the system.
End the run and leave the system filled with 20% ethanol during the storage period.

What is this section about?

This section describes how to replace tubing and connectors, and how to replace the inlet filters.

What should you make sure before starting the replacement of tubing?

Make sure that the entire system tubing is flushed with demineralized water, before starting the replacement of tubing.

When should the inlet filters be replaced?

Replace the inlet filters when required, for example when the visual inspection shows that the filters are clogged or damaged and cleaning is not sufficient.

What items are required for replacing the inlet filters?

Inlet filters
Support nets

Where are the inlet filters mounted?

The inlet filters are mounted on the inlet tubing at the end that will be immersed in the buffer solution.

How should you replace the inlet filter and support net?

Pull off the inlet filter and the support net from the inlet filter holder.
Fit the new support net and inlet filter, and press the filter in position into the inlet filter holder.

When should the tubing and connectors be replaced?

Replace tubing and connectors when required, for example when the tubing is clogged or bent and the flow is obstructed.

What should be done before starting to replace tubing and connectors?

Before starting to replace tubing and connectors, clean the system flow path with DM water, then remove the inlet tubing from water.

What items are required for replacing tubing and connectors?

Tubing and connectors
Tubing cutter

What should you do to replace the pump tubing?

To replace the pump tubing follow the instruction in the ÄKTA start Maintenance Manual.

What should you do to replace the tubing that connects the UV monitor to the Conductivity monitor?

To replace the tubing that connects the UV monitor to the Conductivity monitor use the pre-bent tubing supplied with the system.

How should you replace the tubing and connectors?

Unscrew the connectors, and disconnect the tubing.
If the tubing has labels, remove the labels and use them with the new tubing later. Discard the used tubing and connectors.
Cut the new tubing to the same length as the original tubing. Use a tubing cutter to get a correct right-angle cut.
Put the labels back on the new tubing.
Mount the connectors on the tubing.
For finger tight connectors: Slide the connector onto the tubing.
For tubing connectors 1/8": Slide the connector onto the tubing. Slide the ferrule onto the tubing with the thick end toward the end of the tubing.
Insert the tubing with connector into the port. Make sure to insert the tubing all the way into the bottom of the port.
Tighten the connector fully.

What is this chapter about?

This chapter describes basic troubleshooting and corrective actions for ÄKTA start.

What should be done for a complete list of error and warning messages?

For a complete list of error and warning messages refer to the ÄKTA start Maintenance Manual.

What does this chapter include?

The sections in this chapter describe the basic troubleshooting for ÄKTA start and include a general checklist that should be completed prior to the troubleshooting work. How to generate a System error report for service purposes is also described.

What should be done for replacement of modules and other module specific problems and corrective actions?

For replacement of modules and other module specific problems and corrective actions, refer to the ÄKTA start Maintenance Manual.

What should be done for problems related to the software?

Refer to the UNICORN start User Manual for problems related to the software.

How should you troubleshoot ÄKTA start?

Follow the checklists below, for system, flow path, and purification.
If problems remain, search for solutions in Section 9.2 Basic troubleshooting.
If problems remain after corrective actions, generate a System error report and contact your local GE Service Engineer.

What are the system checks?

Does the instrument display show the ÄKTA start home screen?
Does the ventilation of the instrument function? The fans should be audible at all times when the instrument is switched on. Contact a GE Service Engineer if the fans at the bottom of the instrument stop running.

What are the flow path checks?

Is all tubing connected correctly?
Is there leakage at any of the connections? Tighten the connections if required.
Is any tubing bent or twisted? Adjust the tubing position to make sure that the flow of liquid is smooth, or replace the tubing if required.
Is the buffer inlet tubing immersed in correct buffer solutions?
Are the inlet filters clean? Clean or replace the filters if required.
Is the column connected correctly?

What are the purification checks?

Has the column been cleaned and prepared according to the recommendations in the column instruction manual?
Has the sample been adjusted to binding buffer conditions?
Has the sample been clarified by centrifugation and/or filtration prior to the sample loading?
Are the correct buffers used for the chosen columns and proteins?
Check buffers for precipitations and contamination. Adjust the required temperature wherever needed.
Are the chosen columns suitable for the chosen target proteins?
Have the stable baselines for UV and Conductivity been established?

What should you do if the Instrument display does not show anything?

Check that the Power cord is connected and that the Power Switch is switched on.
Contact a GE Service Engineer if the Instrument display is damaged.
Contact a GE Service Engineer if there is a communication problem, no signals to the display.

What should you do if there are issues with the touch response on the display?

Re-calibrate the Instrument Display. For details, refer to the ÄKTA start Maintenance Manual.

What should you do if the Display color is not correct?

Perform a Display: Color test (see the ÄKTA start Maintenance Manual). If there is a pattern mismatch, contact a GE Service Engineer.

What should you do if the Display Backlight is not working?

Perform Display: Diagnostics and adjust the backlight setting if necessary (see the ÄKTA start Maintenance Manual). If adjustment does not affect the backlight intensity, contact a GE Service Engineer.

What should you do if there is a noisy UV signal, signal drift or instability?

Remove the air bubbles trapped in the UV flow cell by flushing the cell with DM water or buffer. If persistent, clean the UV flow cell.
Check if the signal is still noisy with DM water.
Degas the buffer before use. Use a technique available in the laboratory, such as vacuum degassing or sonication.
Perform Pump wash to remove the air bubbles from the flow path.
Clean the UV flow cell.

What should you do if you see Ghost peaks in the UV signal?

Degas the buffer before use. Use a technique available in the laboratory, such as vacuum degassing or sonication.
Clean the system flow path.
Clean the column according to the column instructions.
Clean the UV flow cell.

What should you do if there is low sensitivity with the UV signal?

Adjust the Switch valve timing.

What should you do if you see waves on the gradient in the UV signal?

Adjust the Switch valve timing.

What should you do if there is a warning 111: UV intensity low?

Mount flow cell securely and re-calibrate the UV module with the flow cell filled with DM water. If the problem persists, replace the UV module.

What should you do if there is a warning 112: UV intensity high?

Mount the flow cell securely, clean and re-calibrate UV module with the flow cell filled with DM water.

What should you do if there is a warning 115: Flush flow cell and mount securely?

Flush the flow cell. Mount the flow cell properly. Re-calibrate the UV module with flow cell filled with DM water.

What should you do if there is a warning 116: UV base lining ignored?

Clean the flow cell thoroughly with DM water. Re-calibrate the UV module with the flow cell filled with DM water.

What should you do if there is an incorrect or unstable Conductivity reading or out of specification conductivity values?

Clean the Conductivity cell.
Check that the Conductivity module cable is connected properly to the connector behind the flow cell.
Check that the Pump functions properly.
Calibrate the temperature sensor.
Check that the column is equilibrated. If necessary, clean the column.
Check the operation of the Buffer valve.
Connect 0.5 mm ID PEEK tubing between the Conductivity Monitor and the Outlet valve.
Contact a GE Service Engineer if there is a failure of component in Main board.

What should you do if there is a baseline drift or noisy signal in the Conductivity?

Remove the air trapped in the flow cell by flushing the cell with DM water or buffer. If persistent, clean the Conductivity flow cell.
Check that the column is equilibrated. If necessary, clean the column.
Check that the pump functions properly.

What should you do if the Conductivity measurement with the same buffer changes over time at constant temperature?

Clean the Conductivity flow cell.
Check that the Pump and the Buffer valve operate properly.

What should you do if you see waves on the gradient in the Conductivity signal?

Adjust Switch valve timing.

What should you do if there is a wrong absolute conductivity value?

Recalibrate the Conductivity flow cell.

What should you do if there are ghost peaks in the gradient profile in the Conductivity signal?

Clean the Conductivity flow cell.
Check for loose tubing connections. Air bubbles pass through the flow cell.

What should you do if there are non-linear gradients or slow response to %B changes?

Check that the Pump functions properly.
Make sure that the tubing is washed properly. Run System cleaning to clean the system flow path.

What should you do if there is an erratic flow?

Remove the air bubbles trapped in the flow path by flushing the flow path with DM water or buffer:

remove the column and re-connect the flow path
perform a manual run with a flow rate of 5 ml/min for about 10 minutes
observe the graph until the pulsations are no longer visible and the curve is stable.

What should you do if there is an inaccurate flow rate (detected by incorrect fraction sizes)?

Calibrate the Pump.
Place the pump tubing in the pump hood maintaining equal distance on both the sides.
Replace the pump tubing.
Fix the pump tubing inside the pump hood and start the run.

What should you do if there is no flow?

Troubleshoot the Pump and if the problem still persists, contact a GE Service Engineer.
Check the tubing connections. Re-tighten or replace if necessary.

What should you do if there is leakage from the Mixer?

Check the tubing connections. Re-tighten or replace if necessary.

What should you do if the fractions collected fall outside the collection tubes?

Make sure that the Bowl is fitted properly on the base.
Check that the dispenser arm is in the dispensing position. Notice the alignment of the markings on the dispenser arm.
Replace the drive sleeve.

What should you do if the Fraction collector is not connected to the instrument?

Connect the Fraction collector to ÄKTA start.

What should you do if the Fraction collector is not set to the home position at the start of the run?

Enable the Fraction collector option from the Settings and service screen.

What should you do if there is a leakage from the valve ports or connections?

Check the tubing connections. Tighten or replace if required.

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