PROTEIN PURIFICATION AND IDENTIFICATION

摘要

The present invention relates to a method for protein purification and identification. More closely the invention relates to a method for protein pre-fractionation and identification resulting in improved yield of identified proteins. The method for pre-fractionation of protein samples, includes the following steps: a) reducing disulphide bridges (S-S bridges) or protecting cysteines in the proteins in the sample; b) loading the sample onto an ion exchange column; c) eluting the sample; d) collecting each fraction from the column separately in air sealed containers devoid of chromatographic media; e) desalting each fraction on a single RPC (reversed phase chromatography) trap column; f) separating each fraction in a second dimension RPC step to obtain further separated proteins which are collected in fractions; and g) identifying the further separated proteins by MS.