HPLC – AKTA CHROMATOGRAPHY
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GE Healthcare - Model
AKTA purifier - Specifications
AKTA purifier is a versatile protein and peptide purification system compatible with any type of chromatographic columns and solvents.
It can operate at maximum flow rate of 10 ml/min with a maximum pressure of 25 MPa, which is sufficient for preparative protein purification on a milligram scale.
The instrument is equipped with a P-903 pump (2 pump modules A and B, including two pump heads each), Monitor UV-900 (able to detect up to three wavelengths simultaneously), Monitor pH/C-900 (measurement of pH and conductivity, pH electrode), and a Frac-920 fraction collector.
The system is connected to a PC and controlled by Unicorn 5.11 software.
Description
AKTA purifier is a liquid chromatography system designed for fast and reliable separations of Peptides, Nucleic acids. Akta Purifier runs all techniques, from µg to mg scale at flow rates 0.001 ml/min – 10 ml/min and pressure 0-25 MPa. With operational simplicity and a wide range of pre=programmed application protocols and method templates.
The liquid flow path includes two pumps each with two pump heads and the pump inlets are immersed in their respective buffers. The buffer solutions are pumped to a mixer by the pump. The flow path continues from the mixer to the injection valve, via an online filter. A sample loop is connected at the injection valve, which can be filled manually using a syringe or through the fill port connected at the injection valve. The flow is then directed through the column, the UV cell in the optical unit and the conductivity cell located below the optical unit and then to the pH monitor. The flow path then continues through the flow restrictor. The flow restrictor generates a constant backpressure to eliminate the risk of air bubbles entering the UV cell. An outlet valve is used to switch the outlet flow to either waste or fraction collection or outlet fraction. The unit is connected to the system with the supporting software, that enables monitoring and analysis of the whole process.
Working Principle
The particle size of Ion exchange columns the Mono Q 5/50 GL( strong Anion exchanger) and Mono S 5/50 GL (strong Cation exchanger) is 10 µm.
Mono Q strong anion exchanger includes the –CH2-N+(CH3)3 charged group with an Ionic capacity of 0.27 - 0.37mmol cl- /ml medium
Mono S strong cation exchanger includes the –CH2-SO3- charged group with Ionic capacity of 0.12 - 0.15mmol H+ /ml medium
Broad flow rate range of 0.01 ml to 100ml.
The system provides the pressure range of 0 to 10 MPa.
Online UV detection of up to three wavelengths in the range of 190-700nm in steps of 1 nm simultaneously.
Online Monitoring of the conductivity of range 1µS/cm to 999.9 mS/cm
Online monitoring of the pH of range 2 to 12.
Fraction Collector options ranging from microlitre plates to large vessels. [It allows collection of up to 175 fractions in 12mm diameter tubes, up to 95 fractions in 10-18 mm tubes, as well as 40 fractions in 30mm diameter tubes.]
Fraction kit enables collection of large volumes of purified sample, thereby improving protein recovery.
The ÄKTA purifier is a fully automated liquid chromatography system designed for method development and research applications.
The system offers automated purification and characterization of peptides and nucleic acids.
It can be used for the high performance purification of fungal glycosidases and nitrilases (ion exchange chromatography, hydrophobic interaction chromatography, gel filtration, chromatofocusing, affinity chromatography and reverses phase chromatography)
Samples should be in liquid form and should be filtered before submission.
Preferred column, and Analysis parameters are to be mentioned in sample submission form
The volume of the sample to be submitted depends on sample load. Minimum volume required to be submitted is 1ml.
Required volume and time of collection of the purified samples to be collected in fraction collector need to be clearly mentioned at the time of sample submission.