New methods of protein purification. Displacement chromatography

摘要

Review discussing a new technique for protein purification, displacement chromatography. Elution in displacement chromatography is carried out by a substance, so called displacer, which has a higher affinity for the chromatographic matrix than any of the adsorbed proteins. The latter are 《pushed》out from the matrix by the displacer and form a displacement train when moving along the column. The component with the weakest affinity moves first and the one with the highest affinity moves last in the train. The concentration of the component in the displacement zone is determined by the interception of the operating line and binding isotherm of the given component. the shape of the displacement zone is close to rectangular. A more powerful resolving force is created especially at high column loadings when nearly all working capacity of the column is used. Independence of component concentration in the displacement zone on the component concentration in the feed allows significant concentrating of the purified protein during displacement chromatography. Examples of application of displacement chromatography for the separation of model protein mixtures and purification of proteins from crude extracts are discussed as well as the methods of column regeneration.