New method for the selective capture of antibodies under physiological conditions

Bioseparation
Luc GuerrierEgisto Boschetti

Abstract

Hydrophobic charge induction chromatography is a recently developed method for protein separation based on the use of dual-mode ligands. They are designed in such a way so as to combine a molecular interaction supported by a mild hydrophobic association effect in the absence of salts. When environmental pH is changed, the ligand becomes ionically charged resulting into the desorption of the protein. This method is applied to the separation of antibodies from ascite fluids and culture supernatants from hybridomas cultured in the presence of fetal bovine serum or in protein free environment. Typically adsorption from cell culture supernatants is accomplished without any pH or ionic strength adjustment; the column is then washed with a typical buffer to eliminate protein impurities. Antibodies are then desorbed using acetate buffer, pH 4. Antibody binding capacity is in the range of 30 mg per ml of resin at 10% breakthrough. Antibody purity varies according to the initial feed stock and can reach values higher than 90% in a single pass. One example of antibody purification process involving hydrophobic charge induction chromatography as a capture step followed by a polishing phase with DEAE Ceramic HyperD is described. Longevity a...Continue Reading

Citations

Aug 8, 2006·Biotechnology Journal·Andrea Buchacher, Günter Iberer
Jan 24, 2007·The FEBS Journal·Pier Giorgio Righetti, Egisto Boschetti
Nov 1, 2017·Current Protein & Peptide Science·Charlotte Cabanne, Xavier Santarelli
Nov 10, 2012·Biotechnology Journal·Kristian KallbergLeif Bulow
Jul 14, 2011·Biotechnology Journal·Caryn L HeldtRuben G Carbonell
Apr 5, 2014·Journal of Molecular Recognition : JMR·Simon J MountfordMilton T W Hearn
Oct 27, 2017·Current Protein & Peptide Science·Atsushi HiranoTomoshi Kameda
Mar 26, 2003·Journal of Chromatography. B, Analytical Technologies in the Biomedical and Life Sciences·An WillemsNico Mertens
Dec 1, 2006·Journal of Chromatography. B, Analytical Technologies in the Biomedical and Life Sciences·Duncan LowNarahari S Pujar
Oct 14, 2008·Protein Expression and Purification·Tsutomu ArakawaDaisuke Ejima
May 2, 2006·Environmental Science & Technology·Miriam M NgundiChris R Taitt

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