Analysis of Non-reduced SDS-PAGE Purity of Recombinant Monoclonal Antibody

Objective To analyze the non-reduced SDS-PAGE purity of recombinant monoclonal antibody(McAb)and exclude the artifacts formed during preparation of test samples.Methods The condition for routine SDS-PAGE was modified based on the structural characters of recombinant McAb molecules.The recombinant McAb samples were treated with various loading buffers under various conditions for SDS-PAGE,and the results were compared.Results Almost all the minor bands appeared during determination of purity of McAb by non-reduced SDS-PAGE were eliminated by addition of alkylation reagent into test sample buffer to block the free sufhydryl groups.The minor bands with high relative molecular mass could be diminished effectively by decreasing the ambient temperature during non-reduced SDS-PAGE.Conclusion The artifacts formed during preparation of test samples due to the disulfide bonds in McAb may be eliminated by modification of condition for non-reduced SDS-PAGE.