Recombinant NY-ESO-1 cancer antigen: production and purification under cGMP conditions |
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| Authors: | Murphy R Green S Ritter G Cohen L Ryan D Woods W Rubira M Cebon J Davis I D Sjolander A Kypridis A Kalnins H McNamara M Moloney M B Ackland J Cartwright G Rood J Dumsday G Healey K Maher D Maraskovsky E Chen Y T Hoffman E W Old L J Scott A M |
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| Affiliation: | Ludwig Institute for Cancer Research, Melbourne Tumour Biology Branch, Austin Hospital, Heidelberg, Australia. roger.murphy@ludwig.edu.au |
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| 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. |
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