Evaluating pre-treatment methods in the isolation and purification of monoclonal IgG antibody

Abstract

In this study, the effects of different sample preparation techniques on the separation of monoclonal antibody IgG I were inve st igated experimentally. Monoclonal IgG I was obtained from hybridoma cell line TB /C3 transfected with bcl-2 carrier plasmid. which was grown in serum-free medium. Three different pre-treatment techniques prior to Protein G affinity chromat ography have been used in order to concentrate and partial purify the monoclonal antibody . The pre-treatments researched in this paper are precipitation of the antibody by ammonium sulfate, dilution of the antibody in the binding buffer of affinity chromatography and ultra filtration through an Amicon Ultra- 15 filter with molecular weight cut-off at 100 kDa. Purification through direct application of the antibody onto the Protein G affinity column without pre-treatments was used as a control method. The results indicate that the ultra filtration through an Amicon filter was an effective method for both concentration and partial purification of the antibodies in laboratory scale.