Abstract
Recombinant DNA-based manufacturing of biologicals has necessitated the development of processes to recover products from very dilute solutions in high yield and purity. Immunoaffinity chromatography is one such method that takes advantage of the specific and reversible binding interaction of antibody with its antigen, which can often result in purification of the target protein in a single step.
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References
Porath J. and Axen R. (1976) Immobilization of enzymes to agar, agarose, and Sephadex supports. Methods Enzymol. 44, 19–45.
Scouten W. H. (1987) A survey of enzyme coupling techniques. Methods Enzymol. 135, 30–65.
Wilchek M., Oka T., and Topper Y. J. (1975) Structure of a soluble super-active insulin is revealed by the nature of the complex between cyanogen-bromide-activated Sepharose and amines. Proc. Nat. Acad. Sci. USA 72, 1055–1058.
Tesser G. I., Fisch H. U., and Schwyzer R. (1974) Limitations of affinity chromatography: solvolytic detachment of ligands from polymeric supports. Helv. Chim.Acta. 57, 1718–1730.
Cuatrecasas P. (1970) Protein purification by affinity chromatography. Derivatizations of agarose and polyacrylamide beads. J. Biol. Chem. 245, 3059–3065.
Cress M. C. and Ngo T. T. (1989) Site specific immobilization of immunoglobulins. Am. Biotechnol. Lab. 7, 16–19.
Orthner C. L., Highsmith F. A., Tharakan J., Madurawe R. D., Morcol T., and Velander W. H. (1991) Comparison of the performance of immunosorbants prepared by site-directed or random coupling of monoclonal antibodies. J. Chromatogr. 558, 55–70.
Gersten D. M. and Marchalonis J. J. (1978) A rapid, novel method for the solid-phase derivatization of IgG antibodies for immune-affinity chromatography. J. Immunol. Methods 127, 215–220.
Schneider C., Newman R. A., Sutherland D. R., Asser U., and Greaves M. F. (1982) One step purification of membrane proteins using a high efficiency immunomatrix. J. Biol. Chem. 257, 10,766–10,769.
Sisson T. H. and Castor C. W. (1990) An improved method for immobilizing IgG antibodies on protein A-agarose. J. Immunol. Methods 127, 215–220.
Stern A. S. and Podlaski F. J. (1993) Increasing the antigen binding capacity of immobilized antibodies. Tech. Protein Chem. 4, 353–360.
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© 2000 Humana Press Inc.
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Podlaski, F.J., Stern, A.S. (2000). Site-Specific Immobilization of Antibodies to Protein G-Derivatized Solid Supports. In: Bailon, P., Ehrlich, G.K., Fung, WJ., Berthold, W. (eds) Affinity Chromatography. Methods in Molecular Biology, vol 147. Humana Press. https://doi.org/10.1007/978-1-60327-261-2_4
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DOI: https://doi.org/10.1007/978-1-60327-261-2_4
Publisher Name: Humana Press
Print ISBN: 978-0-89603-694-9
Online ISBN: 978-1-60327-261-2
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