Kinetic aspects of membrane-based immunoaffinity chromatography.
With a view towards the efficient large-scale purification of recombinant proteins, factors influencing antigen-antibody adsorption kinetics were studied in a model hollow-fiber membrane-based immunosorbent. Non-diffusion-controlled, homogeneous adsorption kinetics are approached in membranes. It is shown that adsorption kinetics, rather than mass transfer, first becomes limiting in membrane-based immunoaffinity chromatography (MIC). Antigen adsorption is not kinetically limited, even at low feed-stream antigen concentrations. Binding efficients approach theoretical values when antibody coupling densities are decreased sufficiently. Antigen breakthrough during adsorption occurs near the membrane's observed binding capacity. The results of these kinetic studies were essential in the development of highly efficient and productive MIC systems for the purification of three recombinantly produced biotherapeutics, interferon-alpha 2a, interleukin-2 and interleukin-2 receptor.
Nachman
M
M
Department of Protein Biochemistry, Roche Research Center, Hoffmann-La Roche Inc., Nutley, NJ 07110.
D000327:Adsorption; D000937:Antigen-Antibody Reactions; D000941:Antigens / Q000276:immunology; D055598:Chemical Phenomena; D002627:Chemistry, Physical; D002846:Chromatography, Affinity; D004058:Diffusion; D007370:Interferon Type I / Q000276:immunology; D000077190:Interferon alpha-2; D016898:Interferon-alpha / Q000276:immunology; D007376:Interleukin-2 / Q000302:isolation & purification; D007700:Kinetics; D008567:Membranes, Artificial; D010084:Oxidation-Reduction; D015375:Receptors, Interleukin-2 / Q000276:immunology / Q000302:isolation & purification; D011994:Recombinant Proteins
D000941:Antigens; D007370:Interferon Type I; D000077190:Interferon alpha-2; D016898:Interferon-alpha; D007376:Interleukin-2; D008567:Membranes, Artificial; D015375:Receptors, Interleukin-2; D011994:Recombinant Proteins