Purification Of Peptide By A Charge Polarity Sequence

Generally, any single protein or peptide can be isolated in >90% purity from any crude extract by the use of two complementary modes of HPLC in sequence. If purity > 98% is desired, it may be necessary to use three modes in sequence.

PEPTIDES :
SCX-RPC : Almost all peptides will be retained by our PolySULFOETHYL A™ material at pH 2.7-3.0 and can be separated by charge differences. The fraction of interest is collected and further resolved by RPC. Peptides which coeluted with the peptide of interest in the SCX run and which presumably have similar charge-to-mass characteristics will be unlikely to have similar polarity as well and will be separated in the second dimension.
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The SCX step should be performed first for the following reasons :
1) Ion-exchange columns have 4x higher binding capacity than a RPC column of the same size.
2) The RPC step can conveniently eliminate the salt from the SCX step.

Column Name : PolySULFOETHYL A™ Columns

Compound Name : Polypeptides