Purification of transferrin by magnetic immunoaffinity beads

Abstract

Immunoaffinity adsorbent for transferrin (Tf) purification was prepared by immobilizing anti-transferrin (Anti-Tf) antibody on magnetic monosizepoly(glycidyl methacrylate) beads, which were synthesized by dispersion polymerization technique in the presence of Fe3 O4 nanopowder and obtained with an average size of 2.0 μm. The magnetic poly(glycidyl methacrylate) (mPGMA) beads were characterized by Fourier transform infrared spectroscopy, swelling tests, scanning electron microscopy, electron spin resonance spectroscopy, thermogravimetric analysis and zeta sizing analysis. The density and swelling ratio of the beads were 1.08 g/cm(3) and 52%, respectively. Anti-Tf molecules were covalently coupled through epoxy groups of mPGMA. Optimum binding of anti-Tf was 2.0 mg/g. Optimum Tf binding from aqueous Tf solutions was determined as 1.65 mg/g at pH 6.0 and initial Tf concentration of 1.0 mg/mL. There was no remarkable loss in the Tf adsorption capacity of immunoaffinity beads after five adsorption-desorption cycles. Tf adsorption from artificial plasma was also investigated and the purity of the Tf molecules was shown with gel electrophoresis studies.

Keywords: Anti-transferrin; Immunoaffinity chromatography; Magnetic beads; Poly(glycidyl methacrylate); Transferrin.