Recombinant NY-ESO-1 cancer antigen: Production and purification under cGMP conditions

R. Murphy, S. Green, G. Ritter, L. Cohen, D. Ryan, W. Woods, M. Rubira, J. Cebon, I. D. Davis, A. Sjolander, A. Kypridis, H. Kalnins, M. McNamara, M. B. Moloney, J. Ackland, G. Cartwright, J. Rood, G. Dumsday, K. Healey, D. MaherE. Maraskovsky, Y. T. Chen, E. W. Hoffman, L. J. Old, A. M. Scott

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32 Citations (Scopus)

Abstract

The cancer-testis antigen, NY-ESO-1, has been engineered into a bacterial expression plasmid which incorporates a His6-tag. The plasmid was transfected into E. coli strain BL21 and Master and Working cell banks generated from this expression system. Three 15-litre fermentations were performed under cGMP (code of Good Manufacturing Practice) conditions and the crude NY-ESO-1 tagged protein isolated as solubilised inclusion bodies. A three-step cGMP chromatography process (immobilised metal affinity, anion exchange, and hydrophobic interaction) was utilised to purify the protein. The purified NY-ESO-1 is being used in early stage human cancer vaccine trials in Australia and the U.S.A.

Original languageEnglish
Pages (from-to)119-134
Number of pages16
JournalPreparative Biochemistry & Biotechnology
Volume35
Issue number2
DOIs
Publication statusPublished - 10 May 2005

Keywords

  • Cancer-testis antigen
  • cGMP
  • NY-ESO-1
  • Recombinant protein

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