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

Andrew M. Clausen, Anuradha Subramanian, Peter W. Carr

Research output: Contribution to journalArticlepeer-review

33 Scopus citations

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. Copyright (C) 1999 Elsevier Science B.V.

Original languageEnglish (US)
Pages (from-to)63-72
Number of pages10
JournalJournal of Chromatography A
Volume831
Issue number1
DOIs
StatePublished - Jan 22 1999
Externally publishedYes

Keywords

  • Monoclonal antibodies
  • Zirconia

ASJC Scopus subject areas

  • Analytical Chemistry
  • Biochemistry
  • Organic Chemistry

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