Antibody purification by concanavalin a affinity chromatography


BERELI N., AKGOL S., Yavuz H., DENIZLI A.

JOURNAL OF APPLIED POLYMER SCIENCE, vol.97, no.3, pp.1202-1208, 2005 (SCI-Expanded) identifier identifier

  • Publication Type: Article / Article
  • Volume: 97 Issue: 3
  • Publication Date: 2005
  • Doi Number: 10.1002/app.21862
  • Journal Name: JOURNAL OF APPLIED POLYMER SCIENCE
  • Journal Indexes: Science Citation Index Expanded (SCI-EXPANDED), Scopus
  • Page Numbers: pp.1202-1208
  • Hacettepe University Affiliated: Yes

Abstract

Concanavalin A (Con A) immobilized poly(2-hydroxyethyl methacrylate) (PHEMA) beads in a spherical form (100-150 mu m 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. (c) 2005 Wiley Periodicals, Inc.