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

Efficient harvest and recovery of high-purity monoclonal antibodies was ach ieved using hydrophobic charge induction chromatography (HCIC). Both simple and complex feedstocks were studied, including protein-free cell culture s upernatant and the clarified/concentrated milk of transgenic goats. Viral c learance studies demonstrated a 4-log reduction of MVM virus (minute virus of mice), along with substantial reduction of DNA content. Sorbent characte rization 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 l yotropic salt. Desorption was accomplished under mild conditions - pH 4.0. At this pH, both ligand and antibody carry a net positive charge, and desor ption occurs on the basis of electrostatic charge repulsion. pH-based contr ol of chromatographic function was demonstrated. Chromatography on this ant ibody-selective HCIC sorbent was evaluated as a cost-effective, process-com patible alternative to affinity chromatography protein A sorbents. (C) 2001 Elsevier Science B.V. All rights reserved.