title: Comparison of hydrophobic charge induction chromatography with affinity chromatography on protein A for harvest and purification of antibodies.
abstract: 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.
authors.affiliation: BioSepra Process Chromatography, Life Technologies, A Division of Invitrogen Corporation, Rockville, MD 20850, USA. wschwart@lifetech.com
authors.forename: W
authors.initials: W
authors.lastname: Schwart
authors.forename: D
authors.initials: D
authors.lastname: Judd
authors.forename: M
authors.initials: M
authors.lastname: Wysocki
authors.forename: L
authors.initials: L
authors.lastname: Guerrier
authors.forename: E
authors.initials: E
authors.lastname: Birck-Wilson
authors.forename: E
authors.initials: E
authors.lastname: Boschetti
mesh_terms: D000911:Antibodies, Monoclonal / Q000302:isolation & purification*; D002846:Chromatography, Affinity / Q000379:methods*; D004591:Electrophoresis, Polyacrylamide Gel; D006863:Hydrogen-Ion Concentration; D009994:Osmolar Concentration; D013205:Staphylococcal Protein A / Q000737:chemistry*
chemical_list: D000911:Antibodies, Monoclonal; D013205:Staphylococcal Protein A