Desorption following immunoaffinity chromatography; generalization of a gentle procedure for desorbing antigen.
Possibilities for generalizing the use of 2 gentle procedures for desorption of antigens in immunoaffinity chromatography were investigated. The procedures involve the use of distilled water as desorbing agent and an interruption period during the desorption procedure. Six proteins of different nature and function (albumins, globulins, enzymes, glycoproteins) and of different origins (seeds, pollen, eggs) were tested with use of the corresponding immune sera. The IgG fractions of the immune sera were immobilized on either activated CNBr-Sepharose or Ultrogel. The procedure was used successfully for all the proteins except one which was denatured on short exposure to distilled water. With the 5 other proteins maximum yield of antigen was obtained by combining the use of distilled water with an interruption period during desorption. Ease of desorption depends on the antigen-antibody system. With antibarley beta-amylase antibodies immobilized either on CNBr-Sepharose or Ultrogel, desorption proved to be easier from Ultrogel.
D000818:Animals; D000936:Antigen-Antibody Complex; D002846:Chromatography, Affinity / Q000379:methods*; D011485:Protein Binding / Q000187:drug effects; D011817:Rabbits; D014867:Water
D000936:Antigen-Antibody Complex; D014867:Water