By Meir Wilchek, Irwin Chaiken (auth.), Pascal Bailon, George K. Ehrlich, Wen-Jian Fung, Wolfgang Berthold (eds.)
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Additional info for Affinity Chromatography: Methods and Protocols
Biochemistry 22, 1621. 56 Schwarz 5. Hale, J. , and Beidler, D. E. (1994) Purification of humanized murine and murine monoclonal antibodies using immobilized metal-affinity chromatography. Anal. Biochem. 222, 29. 6. Al-Mashikhi, S. , and Nakai, S. (1988) Separation of immunoglobulins and lactoferrin from cheese whey by chelating chromatography. J. Dairy Sci. 71, 1747–1755. PAM Affinity Chromatography 57 6 Protein A Mimetic (PAM) Affinity Chromatography Immunoglobulins Purification Giorgio Fassina 1.
All the different supports tested so far maintain the ligand recognition properties for immunoglobulins, even if with different functional capaci- PAM Affinity Chromatography 59 Fig. 1. Amino acid sequence and structural model of PAM (TG 19318). ties. Ligand denaturation does not constitute a problem, such as in the case of protein A, and TG19318 columns can withstand a large array of harsh sanitizing agents with no capacity losses. In addition, the low toxicity of TG19318 and the low molecular weight of the resulting fragments reduces considerably the problems of contamination by leaked ligand, as is the case for protein A.
147: Affinity Chromatography: Methods and Protocols Edited by: P. Bailon, G. K. -J. Fung, and W. , Totowa, NJ 57 58 Fassina tion, are usually required (5,6). Lectin jacalin, isolated from jackfruit seeds (7), binds to IgA and can be conveniently used for the affinity purification of IgA from colostrum or serum (8). However, several aspects limit the use of this lectin for large-scale purification of monoclonal IgA from cell culture supernatants. First, jacalin is a biologically active lectin, being a potent T-cell mitogen and a strong B-cell polyclonal activator (9), thus requiring a careful control for ligand leakage into the purified preparation.
Affinity Chromatography: Methods and Protocols by Meir Wilchek, Irwin Chaiken (auth.), Pascal Bailon, George K. Ehrlich, Wen-Jian Fung, Wolfgang Berthold (eds.)