Resolution of thiol-containing proteins by sequential-elution covalent chromatography.

Elution of complex protein mixtures on a matrix containing reactive disulphide bonds (Thiopropyl-Sepharose 6B, Pharmacia) results in immobilisation of thiol-containing molecules. Specific protein fractions can be displaced from the gel using different low-molecular-weight reducing agents. Thus a single sequential elution can separate and resolve thiol-containing proteins in a rapid and convenient step. The method is illustrated with reference to beef liver thiol : disulphide oxidoreductases.