Purification of monoclonal antibodies from cell culture supernatants usinga modified zirconia based cation-exchange support

A method suitable for the isolation of monoclonal antibodies (Mabs) is desc ribed. The protocol utilizes a zirconia based column modified with ethylene diamine-N,N'-tetra(methylenephosphonic) acid to create a novel cation-excha nge chromatographic support. Initial experiments using a linear salt gradie nt demonstrate the ability of this support to efficiently separate Mab from transferrin and bovine serum albumin in a model matrix. Results of the pur ification of Mab from an actual cell culture supernatant over a range in pr otein concentrations are also shown. Analyses by enzyme-linked immunosorben t assay and gel electrophoresis demonstrate that Mabs can be recovered from a cell culture supernatant at high yield (92-98%) and high purity (>95%) i n a single chromatographic step. (C) 1999 Published by Elsevier Science B.V . All rights reserved.