Comparison of hydrophobic charge induction chromatography with affinity chromatography on protein A for harvest and purification of antibodies

被引:117
作者
Schwartz, W [1 ]
Judd, D
Wysocki, M
Guerrier, L
Birck-Wilson, E
Boschetti, E
机构
[1] Life Technol, Div Invitrogen Corp, BioSepra Proc Chromatog, Rockville, MD 20850 USA
[2] Life Technol, Div Invitrogen Corp, Cell Culture Res & Dev, Rockville, MD 20850 USA
[3] Life Technol, Div Invitrogen Corp, BioSepra Proc Chromatog, F-95800 Cergy, France
[4] Genzyme Transgen, Framingham, MA 01701 USA
关键词
hydrophobic charge induction chromatography; affinity chromatography; preparative chromatography; monoclonal; antibodies; proteins; protein A; immunoglobulins; mercaptoethylpyridine;
D O I
10.1016/S0021-9673(00)01013-X
中图分类号
Q5 [生物化学];
学科分类号
071010 ; 081704 ;
摘要
Efficient harvest and recovery of high-purity monoclonal antibodies was achieved using hydrophobic charge induction chromatography (HCIC). Both simple and complex feedstocks were studied, including protein-free cell culture supernatant and the clarified/concentrated milk of transgenic goats. Viral clearance studies demonstrated a 4-log reduction of MVM virus (minute virus of mice), along with substantial reduction of DNA content. Sorbent characterization studies confirmed that HCIC is based on the pH-dependent behavior of a dual-mode, ionizable ligand. Binding, based on hydrophobic interaction, was achieved under near-physiological conditions, and in the absence of lyotropic salt. Desorption was accomplished under mild conditions - pH 4.0. At this pH, both ligand and antibody carry a net positive charge, and desorption occurs on the basis of electrostatic charge repulsion. pH-based control of chromatographic function was demonstrated. Chromatography on this antibody-selective HCIC sorbent was evaluated as a cost-effective, process-compatible alternative to affinity chromatography protein A sorbents. (C) 2001 Elsevier Science B.V. All rights reserved.
引用
收藏
页码:251 / 263
页数:13
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