Recombinant NY-ESO-1 cancer antigen: production and purification under cGMP conditions.
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.
Murphy
R
R
Ludwig Institute for Cancer Research, Melbourne Tumour Biology Branch, Austin Hospital, Heidelberg, Australia. roger.murphy@ludwig.edu.au
Green
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Ritter
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Cohen
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Ryan
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D
Woods
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W
Rubira
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M
Cebon
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Davis
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ID
Sjolander
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Kypridis
A
A
Kalnins
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McNamara
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Moloney
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MB
Ackland
J
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Cartwright
G
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Rood
J
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Dumsday
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G
Healey
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K
Maher
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D
Maraskovsky
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E
Chen
Y T
YT
Hoffman
E W
EW
Old
L J
LJ
Scott
A M
AM
D000951:Antigens, Neoplasm / Q000096:biosynthesis* / Q000235:genetics / Q000302:isolation & purification* / Q000627:therapeutic use; D001315:Australia; D019496:Cancer Vaccines / Q000096:biosynthesis* / Q000235:genetics / Q000302:isolation & purification*; D004345:Drug Industry / Q000592:standards; D017408:Guidelines as Topic; D006801:Humans; D008565:Membrane Proteins / Q000096:biosynthesis* / Q000235:genetics / Q000302:isolation & purification* / Q000627:therapeutic use; D015202:Protein Engineering / Q000379:methods* / Q000592:standards; D011785:Quality Assurance, Health Care / Q000592:standards; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000302:isolation & purification; D012015:Reference Standards
D000951:Antigens, Neoplasm; C104948:CTAG1B protein, human; D019496:Cancer Vaccines; D008565:Membrane Proteins; D011994:Recombinant Proteins