Antibody purification via affinity membrane chromatography method utilizing nucleotide binding site targeting with a small molecule.

ABSTRACT: Here, we present an affinity membrane chromatography technique for purification of monoclonal and polyclonal antibodies from cell culture media of hybridomas and ascites fluids. The m-NBST method utilizes the nucleotide-binding site (NBS) that is located on the Fab variable domain of immunoglobulins to enable capturing of antibody molecules on a membrane affinity column via a small molecule, tryptamine, which has a moderate binding affinity to the NBS. Regenerated cellulose membrane was selected as a matrix due to multiple advantages over traditionally used resin-based affinity systems. Rituximab was used for proof of concept experiments. Antibody purification was accomplished by first capture of injected samples while running equilibration buffer (50 mM sodium phosphate pH 7.0), followed

SUBMITTER: Mustafaoglu N 

PROVIDER: S-EPMC5245175 | biostudies-literature | 2016 Nov

REPOSITORIES: biostudies-literature