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

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 grad...

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Veröffentlicht in:Journal of Chromatography A 1999-01, Vol.831 (1), p.63-72
Hauptverfasser: Clausen, Andrew M, Subramanian, Anuradha, Carr, Peter W
Format: Artikel
Sprache:eng
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Zusammenfassung: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.
ISSN:0021-9673
DOI:10.1016/S0021-9673(98)00999-6