The ÄKTA prime is designed for simple purification of proteins at laboratory scale. When compared to traditional manual operations, the prime offers significant advantages in speed, capacity and fraction selection. The system is fully compatible with modern pre-packed columns such as HiTrap, Hiprep, HiLoad, RESOURSE, other manufacturers columns and self-packed units. In combination with the pre-installed application and method templates, ÄKTA prime will perform entire purifications steps in a fully automated manner at the touch of a button.
The ÄKTA prime comprises a control system, pump, fraction collector and chart recorder, together with valves for buffer selection, sample injection, gradient formation and flow diversion. The Figure shows the flow path of the Technology Facility system.
The pump delivers liquid with high precision over a wide flow rate range to ensure fast and reproducible purification. A switch valve and mixer are used for gradient formation and a pressure sensor prevents damage to columns. An 8-port buffer selection valve allows automated washing and loading events to be programmed.
Motorized valves ensure that buffer or sample selection, sample loading and fraction collection are performed automatically.
High precision on-line monitoring for UV, conductivity and pH can be obtained. UV detection at 254 and 280nm are offered and a temperature sensor built into the conductivity cell ensures automatic compensation during conductivity and pH measurement.
Fractionation is by fixed volume collection or automatic peak fractionation. Peak fractionation can be based on peak detection using slope sensing. Fraction marks and fraction numbers allow easy identification of fractions and peaks.
Two user selectable signals can be recorded directly during purification i.e. the UV trace (including fraction marks) and the conductivity gradient. Other run data, such as theoretical gradient, can be drawn using a post run procedure. In UV mode, there is no risk of working with incorrect range settings since the data can be re-scaled and re-drawn on the chart recorder.
Due to the nature of the pump and to maximize column lifetime it is essential that all buffers are filtered (0.2 µm) and de-gassed before use with the ÄKTA prime. All facilities for buffer preparation may be found in the Protein Production laboratories if not already available. The machine is generally used in the cold room, but can be relocated at room temperature, however, at least 24 hours notice would be required. It is normal to store the prime in eFebruary 6, 2009or short term storage, respectively. We also recommend that the system be thoroughly washed with water before use.
Suitable samples should also be clarified prior to use with the system. This may be achieved by filtering (0.45 or 0.2 µm) or by centrifugation at 10,000 x g for 20 minutes. Sample loading onto the column is volume dependent and can be performed by either injection into a sample-loop (100 µl to 5 ml) or by direct loading through the pump using the buffer selection valve. The use of a 'Superloop' is also permitted.
Simple purifications e.g. desalting, purification of IgG antibodies and purification of tagged proteins can be performed at the touch of a button using specific columns and the Application templates supplied with the machine. The purification of a His-tagged protein, for instance, can be performed this way, but in addition the Application template will wash, strip and re-charge the column prior to equilibration and purification of the protein. User defined methods can also be programed through the use of Method templates. The Method templates are based on many of the most frequently used techniques i.e. ion-exchange chromatography, hydrophobic interaction, and require the user only to input the information necessary for their specific run. These variables are information like flow rate, gradient length, fraction size etc. Once all the information is entered, the method may be saved and the whole process performed at the touch of a button.
Finally, the Protein Production laboratories would not supply a purification system without the columns to go with it! We have a wide range of columns available and the experience to custom pack resins. For more information on the ÄKTA prime and its capabilities, the types of columns which are available or to book a training session on the machine, please contact Jared Cartwright or Mick Miller in the Protein Production laboratories.