Development of biochromatography material for protein G purification

In this study, IgG-bound macroporous cryogel column packing material has been developed and the effectiveness of this column packing material has been evaluated for Protein G purification from Streptococcus. For this purpose, in the first phase of the study, macroporous poly(HEMA-co-IgG) cryogel have been synthesized by using photosensitive ruthenium-based amino acid monomer, bis(2,2'-bipyridil)MATrp-MATrp-ruthenium(II), according to ANADOLUCA method. Characterization of the prepared cryogel columns has been carried out by swelling tests, SEM and FT-IR analyses. The binding capacity studies for poly(HEMA-co-IgG) have been performed using recombinant Protein G. In the second step of the study, the efficacy of colon has been investigated for real samples. For this purpose, Streptococcus equi bacteria has been cultured, the cell wall has been broken and then, Protein G has been purified using poly(HEMA-co-IgG) cryogel after the appropriate pre-concentration process. In the final step of the study, the molecular weight and purity of purified Protein G have been determined using SDS-PAGE method.