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What is two step purification?

KNAUER Multi Method FPLC System, Lab standard

In the basic set up a Lab standard KNAUER Multi Method FPLC System for all Bio-Chromatography methods is adapted. Next to the column selection valve and the outlet valve no other components must be added to the system. The reinjection position of the outlet valve is connected to the sample pump inlet of the injection valve, so that the syringe port is still accessible for sample injection. The sample is injected via the sample loop of the injection valve. The first peak is collected in the sample loop of the injection valve as well and reinjected onto the second column. 

Consideration: This set up is limited to small sample volumes. The volume of the elution peak of the first column should be equal or even smaller than the injection volume due to diffusion effects that will result in sample loss. 

To overcome the problem of the small sample volume a variation of this set up is to inject the sample via the major pump. For this set up some additional changes must be implemented. The filter cartridge of the pressure senor must be replaced by a dummy and we strongly advise to install an inline filter in front of the column selection valve. 

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