Abstract
Monoclonal antibodies have many applications in biotechnology such as immuno affinity chromatography, immuno diagnostics, immunotherapy, ding targeting, and biosensors among others. For all these applications, homogeneous antibody preparations are needed. Affinity chromatography, which relies on the specific interaction between an immobilized ligand and a particular molecule sought to be purified, is a well-known technique for the purification of proteins from solution. Three different affinity techniques forthe one-step purification of antibodies involving Protein A, thiophilic adsorption, and immobilized metal affinity chromatography (IMAC) are described in this chapter.
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References
Diesenhofer J. (1981) Crystallographic refinement and atomic models of a human Fc fragment and its complex with fragment B of protein A from Staphylococcus aureus at 2.9-and 2.8 A resolution. Biochemistry 20, 2361.
Porath J.. Maisano F.. and Belew M. (1985) Thiophilic adsoprtion-a new method for protein fractionation. FEBS Lett. 185, 306.
Oscarsson S., and Porath J. (1990) Protein chromatography with pyridine-and alkyl-thioether-based agarose adsorbents. J. Chromatogr. 499, 235.
Porath J., and Olin B. (1983) Immobilized metal ion affinity adsorption and immobilized metal ion affinity chromatography of biomaterials. Serum protein affinities for gel-immobilized iron and nickel ions. Biochemistry 22, 1621.
Hale J. E., and Beidler D. E. (1994) Purification of humanized murine and murine monoclonal antibodies using immobilized metal-affinity chromatography. Anal. Biochem. 222, 29.
Al-Mashikhi S. A., Li-Chan E., and Nakai S. (1988) Separation of immunoglobulins and lactoferrin from cheese whey by chelating chromatography. J. Dairy Sci. 71, 1747–1755.
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© 2000 Humana Press Inc
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Schwarz, A. (2000). Affinity Purification of Monoclonal Antibodies. 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_5
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DOI: https://doi.org/10.1007/978-1-60327-261-2_5
Publisher Name: Humana Press
Print ISBN: 978-0-89603-694-9
Online ISBN: 978-1-60327-261-2
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