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Updated: Sep 2, 2026

Generation of Murine Monoclonal Antibodies by Hybridoma Technology
Published on: January 2, 2017
Purification of monoclonal antibodies from cell culture supernatants using a modified zirconia based cation-exchange
A M Clausen1, A Subramanian, P W Carr
1University of Minnesota, Department of Chemistry, Minneapolis 55455, USA.
Abstract:
A method suitable for the isolation of monoclonal antibodies (Mabs) is described. The protocol utilizes a zirconia based column modified with ethylenediamine-N,N'-tetra(methylenephosphonic) acid to create a novel cation-exchange chromatographic support. Initial experiments using a linear salt gradient demonstrate the ability of this support to efficiently separate Mab from transferrin and bovine serum albumin in a model matrix. Results of the purification of Mab from an actual cell culture supernatant over a range in protein concentrations are also shown. Analyses by enzyme-linked immunosorbent assay and gel electrophoresis demonstrate that Mabs can be recovered from a cell culture supernatant at high yield (92-98%) and high purity (> 95%) in a single chromatographic step.
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