Purification of Monoclonal Antibody from Hybridoma Supernatant Using Protein A/G Affinity Chromatography and Comparing UV-Vis. Spectroscopy to Bradford Assay

Abstract

Monoclonal antibodies from hybridoma cell supernatant are used for a variety of medical and research purposes but must be purified before use. Monoclonal antibodies obtained from hybridoma cell supernatant were purified using a protein A/G agarose resin bead affinity chromatography column. Antibody identity was determined using SDS-PAGE. Antibodies recovered from affinity column were quantified using two industry standard methods: UV-Visible Spectroscopy and the Bradford Assay. Quantitative results are discussed.