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.
D000681:Amylases / Q000302:isolation & purification*; D002846:Chromatography, Affinity / Q000379:methods*; D001467:Hordeum; D007163:Immunosorbent Techniques; D011485:Protein Binding; D014867:Water; D001614:beta-Amylase / Q000302:isolation & purification*
D014867:Water; D000681:Amylases; D001614:beta-Amylase