Comparison of hydrophobic charge induction chromatography with affinity chromatography on protein A for harvest and purification of antibodies.
J Chromatogr A
; 908(1-2): 251-63, 2001 Jan 26.
Article
en En
| MEDLINE
| ID: mdl-11218128
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.
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Colección:
01-internacional
Banco de datos:
MEDLINE
Asunto principal:
Proteína Estafilocócica A
/
Cromatografía de Afinidad
/
Anticuerpos Monoclonales
Idioma:
En
Revista:
J Chromatogr A
Año:
2001
Tipo del documento:
Article
País de afiliación:
Estados Unidos