Strategy for the immobilization of monoclonal antibodies on solid-phase supports. 1988

R S Matson, and M C Little
Bio-Rad Laboratories, Richmond, CA 94804.

Using matrices based upon Affi-Gel and Affi-Prep, we have examined conditions during the immobilization of antibodies (immunoglobulin G, IgG) that influence the performance of immunosorbents. Such conditions include: coupling pH, coupling kinetics, antibody density on the immunosorbent and the activation chemistries utilized for the immobilization process. These studies have shown that the capacity for antigen does not increase with increased antibody coupling efficiency. Presumably, increased coupling times or efficiencies lead to multi-site attachment of the antibody to the matrix, thereby causing inactivation. Immunosorbents containing low densities of IgG were found to have greater capacity for antigen on a per mole IgG basis. This suggests steric crowding of antigen at high antibody density. Finally, immunosorbents prepared through IgG carbohydrate linkages (oriented coupling) show dramatic increases in antigen capacity over those prepared by stochastic (random) coupling through IgG primary amino groups. A combination of low IgG density and oriented coupling of the IgG via the carbohydrate moiety may represent the best strategy for the preparation of immunosorbents.

UI MeSH Term Description Entries
D007700 Kinetics The rate dynamics in chemical or physical systems.
D002846 Chromatography, Affinity A chromatographic technique that utilizes the ability of biological molecules, often ANTIBODIES, to bind to certain ligands specifically and reversibly. It is used in protein biochemistry. (McGraw-Hill Dictionary of Scientific and Technical Terms, 4th ed) Chromatography, Bioaffinity,Immunochromatography,Affinity Chromatography,Bioaffinity Chromatography
D006863 Hydrogen-Ion Concentration The normality of a solution with respect to HYDROGEN ions; H+. It is related to acidity measurements in most cases by pH pH,Concentration, Hydrogen-Ion,Concentrations, Hydrogen-Ion,Hydrogen Ion Concentration,Hydrogen-Ion Concentrations
D000911 Antibodies, Monoclonal Antibodies produced by a single clone of cells. Monoclonal Antibodies,Monoclonal Antibody,Antibody, Monoclonal
D000937 Antigen-Antibody Reactions The processes triggered by interactions of ANTIBODIES with their ANTIGENS. Antigen Antibody Reactions,Antigen-Antibody Reaction,Reaction, Antigen-Antibody,Reactions, Antigen-Antibody
D000941 Antigens Substances that are recognized by the immune system and induce an immune reaction. Antigen

Related Publications

R S Matson, and M C Little
June 2008, The Analyst,
R S Matson, and M C Little
February 2018, Nature communications,
R S Matson, and M C Little
January 2007, Nucleosides, nucleotides & nucleic acids,
R S Matson, and M C Little
January 1984, Methods in molecular biology (Clifton, N.J.),
R S Matson, and M C Little
October 1985, Journal of immunological methods,
R S Matson, and M C Little
January 2000, Methods in molecular biology (Clifton, N.J.),
R S Matson, and M C Little
February 1992, Journal of chromatography,
R S Matson, and M C Little
January 1986, Voprosy virusologii,
R S Matson, and M C Little
October 1981, Medical laboratory sciences,
R S Matson, and M C Little
December 1987, Biotechnology and applied biochemistry,
Copied contents to your clipboard!