title: Recombinant NY-ESO-1 cancer antigen: production and purification under cGMP conditions.
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.
authors.affiliation: Ludwig Institute for Cancer Research, Melbourne Tumour Biology Branch, Austin Hospital, Heidelberg, Australia. roger.murphy@ludwig.edu.au
authors.forename: R
authors.initials: R
authors.lastname: Murphy
authors.forename: S
authors.initials: S
authors.lastname: Green
authors.forename: G
authors.initials: G
authors.lastname: Ritter
authors.forename: L
authors.initials: L
authors.lastname: Cohen
authors.forename: D
authors.initials: D
authors.lastname: Ryan
authors.forename: W
authors.initials: W
authors.lastname: Woods
authors.forename: M
authors.initials: M
authors.lastname: Rubira
authors.forename: J
authors.initials: J
authors.lastname: Cebon
authors.forename: I D
authors.initials: ID
authors.lastname: Davis
authors.forename: A
authors.initials: A
authors.lastname: Sjolander
authors.forename: A
authors.initials: A
authors.lastname: Kypridis
authors.forename: H
authors.initials: H
authors.lastname: Kalnins
authors.forename: M
authors.initials: M
authors.lastname: McNamara
authors.forename: M B
authors.initials: MB
authors.lastname: Moloney
authors.forename: J
authors.initials: J
authors.lastname: Ackland
authors.forename: G
authors.initials: G
authors.lastname: Cartwright
authors.forename: J
authors.initials: J
authors.lastname: Rood
authors.forename: G
authors.initials: G
authors.lastname: Dumsday
authors.forename: K
authors.initials: K
authors.lastname: Healey
authors.forename: D
authors.initials: D
authors.lastname: Maher
authors.forename: E
authors.initials: E
authors.lastname: Maraskovsky
authors.forename: Y T
authors.initials: YT
authors.lastname: Chen
authors.forename: E W
authors.initials: EW
authors.lastname: Hoffman
authors.forename: L J
authors.initials: LJ
authors.lastname: Old
authors.forename: A M
authors.initials: AM
authors.lastname: Scott
mesh_terms: 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
chemical_list: D000951:Antigens, Neoplasm; C104948:CTAG1B protein, human; D019496:Cancer Vaccines; D008565:Membrane Proteins; D011994:Recombinant Proteins