ӒKTA pure™ 25 T

ÄKTA pure protein purification system: Sample automation

The modular design allows the user to customize ÄKTA pure in multiple ways. The system is always delivered with the core modules of the selected configuration, but one or more optional modules may be added to the flow path.
The table below contains information on the core modules and the optional modules of ÄKTA pure 25 and ÄKTA pure 150. 

The valves for ÄKTA pure 25 and ÄKTA pure 150 are compatible with both systems but for the best performance the specific valve type should be used.

Note: All valve kits include the necessary tubing.

Module	Label in ÄKTA pure 25	Label in ÄKTA pure 150
System pump A	P9 A	P9H A
System pump B	P9 B	P9H B
Pressure monitor	R9	R9
Mixer	M9	M9
Injection valve	V9-Inj	V9H-Inj
Inlet valve A	V9-IA	V9H-IA
Inlet valve B	V9-IB	V9H-IB
Inlet valve AB	V9-IAB	V9H-IAB
Inlet valve IX	V9-IX	V9H-IX
Sample inlet valve	V9-IS	V9H-IS
Mixer valve	V9-M	V9H-M
Loop valve	V9-L	V9H-L
Column valves	V9-C V9-Cs	V9H-C V9H-Cs
pH valve	V9-pH	V9H-pH
Outlet valves	V9-O V9-Os	V9H-O V9H-Os
Versatile valve	V9-V	V9H-V
UV monitors	U9-L U9-M	U9-L U9-M
Conductivity monitor	C9	C9
External air sensor	L9-1.5 L9-1.2	L9-1.5 L9-1.2
Fraction collectors	F9-C F9-R	F9-C F9-R
I/O box	E9	E9
Sample pump	S9	S9H

 

Location and illustration of UV monitor U9-M, UV monitor U9-T, and UV monitor U9-L

The illustration below shows the location of UV monitor U9-M with a detailed view of the monitor unit and detector. The monitor unit is labelled U9-M and the detector U9-D.

Note: When UV monitor U9-M is used, the entire Multi-module panel shown in the illustration is replaced by U9-M.

The illustration below also shows the recommended location of UV monitor U9-L and UV monitor U9-T, together with a detailed view of the monitor .monitor. The UV monitors U9-L and U9-T require that the multi-module panel is installed.

Part	Description
1	Inlet
2	UV flow cell. Three different path lengths are available: 0.5 mm, 2 mm (default) and 10 mm
3	UV detector
4	Outlet
5	Inlet
6	UV monitor U9-L and UV monitor U9-T
7	UV flow cell. Two different path lengths are available: 2 mm (default) and 5 mm
8	Outlet
9	UV detector

 

Function of UV monitor U9-L

The UV monitor U9-L measures the UV absorbance at the fixed wavelength of 280 nm. It is not possible to vary the wavelength, or turn on or off the U9-L monitor. This is therefore not shown in the Phase Properties pane in Method Editor.

For information on how to install UV monitor U9-L, please refer to Section 4.10 Install UV monitor U9-L, in the system handbook.

 

Function of UV monitor U9-T

The UV monitor U9-T measures the UV absorbance at the fixed wavelengths of 260 and 280 nm. It is not possible to vary the wavelength or turn on or off the U9-T monitor on and off. This is therefore not shown in the Phase Properties pane in Method Editor.

For information on how to install UV monitor U9-T, please refer to Section 4.10 Install UV monitor U9-T, in the system handbook.

 

Using two UV monitors

The UV monitor U9-T module has two configurations: UV monitor U9-T and UV monitor U9-T, 2nd. The configuration is defined by the module's Node ID. It is possible to use two UV monitors for ÄKTA pure™ in the following combinations:
• UV monitor U9-M together with UV monitor U9-L and or U9-T, 2nd

• UV monitor U9-L together with UV monitor U9-L, 2nd

UV monitor U9-T, 2nd can be located anywhere in the flow path and is therefore shown in the Process Picture as a component without a fixed place. This means that it is possible to place U9-T, 2nd before the other UV monitor in the flow path.
Note: If U9-T, 2nd is placed on the high-pressure side of the column, pressure limits have tomust be considered.

Location and illustration of the pH monitor
The illustration below shows the location of the pH flow cell and a pH electrode installed in the pH valve.

Part	Description
1	pH flow cell
2	pH electrode

 

The illustration below shows the location, together with a detailed view of Mixer M9.

Part	Description
1	Outlet
2	Mixer chamber
3	Inlet

 

Select Mixer chamber
To obtain a homogeneous buffer composition, it is important to use a Mixer chamber suitable for the flow rate of the method. The tables below show what Mixer chambers to use in ÄKTA pure at different flow rates.
If the liquids are difficult to mix, use a larger Mixer chamber to achieve optimal mixing. However, note that a larger Mixer chamber distorts and delays the gradient.
Three different Mixer chambers are available for ÄKTA pure 25. Their volumes are: 0.6 ml, 1.4 ml (mounted at delivery), and 5 ml.

Mixer chamber volume [ml]	Flow rate [ml/min], Binary gradient
0.6	0.1-5
1.4	0.5-10
5	5-25

 

CAUTION
Risk of explosion. Do not use Mixer chamber 15 ml with an ÄKTA pure 25 configuration.
The maximum pressure for Mixer chamber 15 ml is 5 MPa.

 

Four different Mixer chambers are available for ÄKTA pure 150. Their volumes are: 0.6 ml, 1.4 ml
(mounted at delivery), 5 ml (included in delivery), and 15 ml.

 

Mixer chamber volume [ml]	Flow rate [ml/min], Binary gradient
0.6	0.5-5
1.4	0.5-15
5	2-50
15	15-150

Note: At low flow rates (below 0.2 ml/min) a gradient of sufficient quality may be achieved with the mixer bypassed.
Note: For the highest flow rates, use the 5 ml mixer for ÄKTA pure 25 and the 15 ml mixer for ÄKTA pure 150. The larger chamber volume minimizes potential pump disturbances.
Note: The 1.4 ml mixer might work up to 25 ml/min provided that the buffers are easily mixed.

Note: O-ring 13.1 x 1.6 mm 28-9535-45 is for use with Mixer chamber 0.6, 1.4 and 5 ml.

Note: O-ring 22.1 x 1.6 mm 28-9818-57 is for use with Mixer chamber 15 ml.

Note: O-ring 13.1 x 1.6 mm 29-0113-26 (high resistant) can be used as an alternative to 29-9535-45.

Illustration of Fraction collector F9-R, connector panel, base unit and delivery arm.

Part	Function
1	Lock knob
2	Delivery arm
3	Tubing connector
4	Tube sensor
5	Collection tubes
6	Tube rack
7	Base unit
8	Node switch
9	UniNet-9 communication and power supply
10	Central spindle
11	Drive sleeve
12	Delivery arm
13	Delivery arm
14	Tube adjustment cavity
15	Sensor control
16	Tubing holder
17	Tubing holder nut
18	Exposed tubing end length

 

Front view

Part	Description
1	Fractionation indicator Symbol indicating that fractionation is ongoing. Do not open the door while the indicator is lit.
2	Door
3	Window
4	Door handle
5	Tubing connector for outlet valve tubing
6	Vents

 

Rear view

Part	Description
1	Vents
2	UniNet-9 D-type connector (for communication and power supply)
3	Waste tube

 

Interior

Part	Description
1	Fractionation arm guide rail
2	Fractionation arm main rail
3	Lamp
4	Tubing guide
5	Tubing connection
6	Dispenser head
7	Fractionation arm
8	Tubing guide
9	Height exclusion bar
10	Waste funnel
11	Waste tube
12	Tray catch
13	Waste groove, in case of overflow
14	Tray guides

 

Dispenser head

Part	Description
1	Dispenser head
2	Nozzle
3	Dispenser head cover
4	Accumulator (back part of Dispenser head)
5	Drop sync sensor
6	Type code reader

 

Up to four external air sensors can be added to ÄKTA pure, and there are two different versions to choose from. They differ in internal diameter and optimal position on the instrument.

The air sensors can be attached to the instrument using the rails and holders. No dummy modules need to be removed.
In addition to be used for preventing air from entering the system, the external air sensors can be used together with System pump A to load the entire sample volume.

Air sensor L9-1.5
L9-1.5 has a 1.5 mm inner diameter and is designed for i.d. 1.6 mm FEP tubing at the low pressure side before pumps. It is installed in the flow path before the system pumps and is used to prevent air entering the valve.

Air sensor L9-1.2
L9-1.2 has a 1.2 mm inner diameter and is designed for o.d. 1/16" tubing at the high pressure side after the pumps. It is installed after the injection valve and is used to prevent air entering the column.

 

Tubing connections 

Air sensor L9-1.5

Connection between	Tubing	Connector
L9-1.5 and inlet valves	FEP, o.d. 1/8", 1.6 mm	Tubing connector, 5/16" + Ferrule (yellow), 1/8"

Note: When the air sensor is installed before System pump A, use tubing with a minimum length of 18 cm between the valve located before System pump A and the external air sensor when loading the sample at maximum flow rate, to be sure that air does not reach System pump A. Shorter tubing can be used at lower flow rates.

Air sensor L9-1.2

Connection between	Tubing	Connector
V9-Inj and L9-1.2	PEEK, o.d. 1/16"	Fingertight connector 1/16"
L9-1.2 and V9-C/V9-Cs/the connected column	PEEK, o.d. 1/16"	Fingertight connector 1/16"

 

Location and illustration of Conductivity monitor C9n
The illustration below shows the recommended location of Conductivity monitor C9n, together with a detailed view of the monitor. The monitor is labelled C9.

 

Part	Description
1	Inlet
2	Conductivity flow cell
3	Conductivity monitor C9n
4	Outlet