title: Process development and production of cGMP grade Melan-A for cancer vaccine clinical trials.
abstract: Melan-A is a cancer testis antigen commonly found in melanoma, and has been shown to stimulate the body's immune response against cancerous cells. We have developed and executed a process utilizing current good manufacturing practices (cGMP) to produce the 6 times-His tagged protein in C41DE3 Escherichia coli for use in Phase I clinical trials. Approximately 11 g of purified Melan-A were produced from a 20 L fed-batch fermentation. Purification was achieved through a three column process utilizing immobilized metal affinity, anion exchange, and cation exchange chromatography with a buffer system optimized for low-solubility, high LPS binding capacity proteins. The host cell proteins, residual DNA, and endotoxin concentration were well below limits for a prescribed dose with a final purity level of 91%.
authors.affiliation: Department of Biomedical Engineering, Cornell University, Ithaca, NY 14853, USA. Electronic address: clb267@cornell.edu.
authors.forename: Cameron L
authors.initials: CL
authors.lastname: Bardliving
authors.forename: Adam J
authors.initials: AJ
authors.lastname: Lowe
authors.forename: Chung-Jr
authors.initials: CJ
authors.lastname: Huang
authors.forename: Laura
authors.initials: L
authors.lastname: Manley
authors.forename: Gerd
authors.initials: G
authors.lastname: Ritter
authors.forename: Lloyd
authors.initials: L
authors.lastname: Old
authors.forename: Carl A
authors.initials: CA
authors.lastname: Batt
mesh_terms: D035843:Biomedical Research; D019496:Cancer Vaccines; D002626:Chemistry, Pharmaceutical; D002852:Chromatography, Ion Exchange; D005285:Fermentation; D006639:Histidine / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D058965:MART-1 Antigen / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D055550:Protein Stability; D011993:Recombinant Fusion Proteins / Q000737:chemistry / Q000235:genetics / Q000302:isolation & purification* / Q000378:metabolism*; D015203:Reproducibility of Results
chemical_list: D019496:Cancer Vaccines; D058965:MART-1 Antigen; D011993:Recombinant Fusion Proteins; C033223:polyhistidine; D006639:Histidine
keywords: CT-antigen; MART-1; Melan-A; Process development; cGMP