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

2005

Murphy, R

Green, S

Ritter, G

Cohen, L

Ryan, D

Woods, W

Rubira, M

Cebon, Jonathan S

Davis, Ian D

Sjolander, A

Kypridis, A

Kalnins, Renate M

McNamara, M

Moloney, M B

Ackland, J

Cartwright, Glenn A

Rood, J

Dumsday, G

Healey, K

Maher, D

Maraskovsky, Eugene

Chen, Y T

Hoffman, E W

Old, Lloyd J

Scott, Andrew M

Journal Article

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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Preparative Biochemistry & Biotechnology; 35(2): 119-34

Preparative biochemistry & biotechnology