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Adsorption equilibrium in immuno‐affinity chromatography with polyclonal and monoclonal antibodies
Author(s) -
Sada Eizo,
Katoh Shigeo,
Sukai Kiyoshi,
Tohma Masaaki,
Kondo Akihiko
Publication year - 1986
Publication title -
biotechnology and bioengineering
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.136
H-Index - 189
eISSN - 1097-0290
pISSN - 0006-3592
DOI - 10.1002/bit.260281007
Subject(s) - polyclonal antibodies , monoclonal antibody , chemistry , chaotropic agent , elution , affinity chromatography , sepharose , chromatography , antibody , antigen , adsorption , langmuir adsorption model , ionic strength , affinities , monoclonal , ligand (biochemistry) , biochemistry , enzyme , biology , organic chemistry , receptor , aqueous solution , immunology
The effects of pH, ionic strength, anion species, and antibody concentration on the adsorption equilibrium between immobilized antibodies and antigens were studied by use of anti‐BSA, anti‐HSA, anti‐BlgG, and monoclonal anti‐HSA coupled to Sepharose 4B. The polyclonal antibodies possessed average binding affinities of the order of 10 8 M −1 , and the heterogeneity was accounted for by assuming a normal distribution of the free energy of antibody–antigen combination. The monoclonal antibody, on the other hand, showed a homogeneous affinity of the Langmuir type. Bound antigens could be eluted by lowering pH or adding a chaotropic anion, and their purity was very high. The antibody ligand was sufficiently stable for repeated use.