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Antibody purification by concanavalin A affinity chromatography
Author(s) -
Bereli Nilay,
Akgöl Sinan,
Yavuz Handan,
Denizli Adil
Publication year - 2005
Publication title -
journal of applied polymer science
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.575
H-Index - 166
eISSN - 1097-4628
pISSN - 0021-8995
DOI - 10.1002/app.21862
Subject(s) - adsorption , chromatography , (hydroxyethyl)methacrylate , aqueous solution , concanavalin a , chemistry , methacrylate , suspension polymerization , human serum albumin , affinity chromatography , covalent bond , polymerization , immunoglobulin g , polymer chemistry , nuclear chemistry , polymer , organic chemistry , antibody , biochemistry , in vitro , enzyme , immunology , biology
Concanavalin A (Con A) immobilized poly(2‐hydroxyethyl methacrylate) (PHEMA) beads in a spherical form (100–150 μm in diameter) were used for the affinity chromatography purification of human immunoglobulin G (IgG) from aqueous solutions and human plasma. PHEMA adsorbents were prepared by suspension polymerization. Bioligand Con A was then immobilized by covalent binding onto PHEMA beads. The maximum IgG adsorption on the PHEMA/Con A beads was observed at pH 6.0. The nonspecific IgG adsorption onto the plain PHEMA adsorbents was very low (ca. 0.17 mg/g). Higher adsorption values (up to 54.3 mg/g) were obtained when the PHEMA/Con A beads were used from aqueous solutions. A higher adsorption capacity was observed for human plasma (up to 69.4 mg/g) with a purity of 82.5%. The adsorption capacities of other blood proteins were 2.0 mg/g for fibrinogen and 4.2 mg/g for albumin. The total protein adsorption was determined to be 76.0 mg/g. IgG molecules could be repeatedly adsorbed and desorbed with the PHEMA/Con A beads without noticeable loss in the IgG adsorption capacity. © 2005 Wiley Periodicals, Inc. J Appl Polym Sci 97: 1202–1208, 2005