Synthesis and Characterization of Macroporous Poly Acrylamide-Methacrylamido Histidine Cryogels and Their Use in Antibody Purification

Year 2008, Volume: 36 Issue: 1, 61 - 76, 01.01.2008

İlker Koç Nilay Bereli Gözde Baydemir Igor Yu Galaev Adil Denizli

Abstract

Poly acrylamide-methacrylamidohistidine [poly AAm-MAH ] cryogel was prepared by bulk polymerization whichproceeds in aqueous solution of monomers frozen inside a syringe column cryo-polymerization . After thawing, themonolithic cryogel contains a continuous polymeric matrix having interconnected pores of 10-100 µm size. Poly AAmMAH cryogel was characterized by swelling studies, FTIR and scanning electron microscopy. The equilibrium swellingdegree of the poly AAm-MAH monolithic cryogel was 6.97 g H2O/g cryogel. Poly AAm-MAH cryogel was used in theadsorption/desorption of immunoglobulin G IgG from aqueous solutions and human plasma. The non-specificadsorption of IgG was very low 0.22 mg/g . The maximum amount of IgG adsorption from aqueous solution in HEPESbuffer was 32.879 mg/g at pH 7.4. Higher IgG adsorption value was obtained from human plasma up to 100 mg/g .Desorption of IgG from poly AAm-MAH cryogel was obtained using 20 mM Acetate buffer containing 1.0 M NaCl atpH 4. It was observed that IgG could be repeatedly adsorbed and desorbed with poly AAm-MAH cryogel withoutsignificant loss in the adsorption capacity

Keywords

Affinity chromatography, cryogel, Protein Purification, IgG

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