Immunoaffinity chromatography of proteins. A gentle and simple desorption procedure

A gentle procedure for desorption in immunoaffinity chromatography was investigated. Barley beta-amylase absorbed on the Sepharose-immobilized IgG fraction of the anti-barley beta-amylase immune serum was used in this study. Elution with water did not desorb the antigen by using a continuous elution procedure. However, desorption with water became effective when the elution procedure involved an interruption of a few hours. Several factors of the desorption in these conditions were further investigated. At best, up to 35% of the absorbed antigen could be desorbed in one fraction. Preliminary assays carried out with another plant protein and its corresponding antibodies provided similar results.