Antibody purification with protein A attached supermacroporous poly(hydroxyethyl methacrylate) cryogel

• Published in: Biochemical engineering journal Vol. 45; no. 3; pp. 201 - 208
• Main Authors: Alkan, Hüseyin, Bereli, Nilay, Baysal, Zübeyde, Denizli, Adil
• Format: Journal Article
• Language: English
• Published: Amsterdam Elsevier B.V 15-08-2009; Elsevier
• Subjects: Antibody purification; Biological and medical sciences; Biotechnology; Cryogels; Fundamental and applied biological sciences. Psychology; IgG; PHEMA; Protein A; PHEMA; IgG; Protein A; Cryogels; Antibody purification; Immunoglobulins; Purification; Antibody; Cryogel; Protein
• ISSN: 1369-703X; 1873-295X
• DOI: 10.1016/j.bej.2009.03.013

Immunoglobulin G (IgG) purification from human plasma with protein A attached supermacroporous poly(hydroxyethyl methacrylate) [PHEMA] cryogel has been studied. PHEMA cryogel was prepared by bulk polymerization which proceeds in aqueous solution of monomer frozen inside a plastic syringe (cryo-polymerization). After thawing, the PHEMA cryogel contains a continuous matrix having interconnected pores of 10–200
μm size. Protein was covalently attached onto the PHEMA cryogel via cyanogen bromide (CNBr) activation. The maximum IgG adsorption on the PHEMA/protein A cryogel was found to be 83.2
mg/g at pH 7.4 from aqueous solutions. The non-specific IgG adsorption onto the PHEMA cryogel was about 0.38
mg/g. The macropore size of the cryogel makes it possible to process blood cells without blocking the column. Higher adsorption capacity was observed from human plasma (up to 88.1
mg/g). Adsorbed IgG was eluted using 0.1
M glycine–HCl buffer (pH 3.5) with a purity of 85%. PHEMA–protein A cryogel was used for repetitive adsorption/desorption of IgG without noticeable loss in IgG adsorption capacity after 10 cycles. PHEMA–protein A cryogel showed several advantages such as simpler preparation procedure, good selectivity for IgG purification from human plasma and good stability throughout repeated adsorption–desorption cycles.