Affinity precipitation of proteins

Journal of Molecular Recognition : JMR
M N GuptaB Mattiasson

Abstract

Affinity precipitation is being studied as a technique to be introduced at an early stage of downstream processing for the selective isolation of proteins. The technique utilizes a heterobifunctional ligand, which, in addition to having affinity for the target protein(s), possesses another function for controlling precipitation. The latter component is comprised of a polymer which can be made reversibly soluble and insoluble by altering a specific parameter such as pH or temperature. Different polymers of natural and synthetic origin have been used for this purpose. The soluble form of the ligand is used for the affinity binding step and precipitation is induced for obtaining separation of the affinity complex. Some of the polymers used in this laboratory include chitosan, alginate, Eudragit S-100 (copolymer of methacrylic acid and methyl methacrylate) and polyethyleneimine. Chitosan and alginate served as natural ligands for wheat germ agglutinin and pectinase, respectively. The aromatic dye Cibacron Blue 3GA coupled to Eudragit S 100 and polyethyleneimine way used for the affinity precipitation of some model enzymes such as lactate dehydrogenase and alcohol dehydrogenase. As prior removal of cell debris, etc., is essential fo...Continue Reading

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Citations

Nov 14, 2006·Analytical Biochemistry·Kusum SolankiMunishwar N Gupta
Sep 24, 1999·Biochimie·M DanielukJ Bandorowicz-Pikuła
May 28, 2003·Journal of Chromatography. B, Analytical Technologies in the Biomedical and Life Sciences·Frank Hilbrig, Ruth Freitag
Oct 22, 2010·Biotechnology and Applied Biochemistry·Om Prakash, Nivedita Jaiswal
Feb 11, 2015·International Journal of Molecular Sciences·Peter Feist, Amanda B Hummon
Dec 18, 2001·Journal of Molecular Recognition : JMR·S TeotiaM N Gupta
Jun 7, 2015·Journal of Materials Chemistry. B, Materials for Biology and Medicine·Dan-Dan ZhangJian-Hua Wang
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Aug 8, 2015·Applied Biochemistry and Biotechnology·Zhaoyang DingXuejun Cao
Jan 29, 2000·Protein Expression and Purification·A SharmaM N Gupta
Apr 21, 2006·Biochimica Et Biophysica Acta·Kalyani MondalMunishwar N Gupta
Dec 23, 2011·Bioconjugate Chemistry·Katherine A MiricaGeorge M Whitesides
Aug 22, 2021·Journal of Chromatography. B, Analytical Technologies in the Biomedical and Life Sciences·Ligen DasAnanda Gupta
May 10, 2008·Journal of Chromatography. a·Smita RaghavaMunishwar N Gupta
Nov 28, 2001·Analytical Chemistry·A BossiA P Turner

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