title: Expression and purification of cGMP grade NY-ESO-1 for clinical trials.
abstract: NY-ESO-1 is a cancer testis antigen expressed in numerous cancers. Initial tests have shown its efficacy as a cancer vaccine, stimulating the body's own immune response against the invading tumor. To produce enough material for phase I clinical trials, a process using current good manufacturing practices to produce clinical grade material was developed and executed. His-tagged NY-ESO-1 was expressed in C41DE3 Escherichia coli under control of the T-7 promoter. NY-ESO-1 was produced in a 20 L fed-batch fermentation utilizing a pH-stat control scheme. The protein was then purified from inclusion bodies using a three-column process that achieved a yield of over 3.4 g and endotoxin below the detection limit of 0.005 EU/μg protein.
authors.affiliation: Graduate Field of Microbiology, Cornell University, Ithaca, NY 14853, USA. ajl248@cornell.edu
authors.forename: Adam J
authors.initials: AJ
authors.lastname: Lowe
authors.forename: Cameron L
authors.initials: CL
authors.lastname: Bardliving
authors.forename: Chung-Jr
authors.initials: CJ
authors.lastname: Huang
authors.forename: Leonardo M
authors.initials: LM
authors.lastname: Teixeira
authors.forename: Leonardo M
authors.initials: LM
authors.lastname: Damasceno
authors.forename: Kyle A
authors.initials: KA
authors.lastname: Anderson
authors.forename: Gerd
authors.initials: G
authors.lastname: Ritter
authors.forename: Lloyd J
authors.initials: LJ
authors.lastname: Old
authors.forename: Carl A
authors.initials: CA
authors.lastname: Batt
mesh_terms: D000951:Antigens, Neoplasm / Q000096:biosynthesis* / Q000302:isolation & purification; D019496:Cancer Vaccines / Q000096:biosynthesis*; D002986:Clinical Trials as Topic / Q000379:methods; D003001:Cloning, Molecular / Q000379:methods; D004731:Endotoxins / Q000032:analysis; D004926:Escherichia coli / Q000235:genetics; D006801:Humans; D008297:Male; D008565:Membrane Proteins / Q000096:biosynthesis* / Q000302:isolation & purification; D013737:Testis / Q000276:immunology*
chemical_list: D000951:Antigens, Neoplasm; C104948:CTAG1B protein, human; D019496:Cancer Vaccines; D004731:Endotoxins; D008565:Membrane Proteins