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%.
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Authors | Cameron L Bardliving, Adam J Lowe, Chung-Jr Huang, Laura Manley, Gerd Ritter, Lloyd Old, Carl A Batt |
Journal | Protein expression and purification
(Protein Expr Purif)
Vol. 92
Issue 2
Pg. 171-82
(Dec 2013)
ISSN: 1096-0279 [Electronic] United States |
PMID | 24045055
(Publication Type: Journal Article)
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Copyright | Copyright © 2013 Elsevier Inc. All rights reserved. |
Chemical References |
- Cancer Vaccines
- MART-1 Antigen
- Recombinant Fusion Proteins
- polyhistidine
- Histidine
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Topics |
- Biomedical Research
- Cancer Vaccines
- Chemistry, Pharmaceutical
- Chromatography, Ion Exchange
- Fermentation
- Histidine
(chemistry, genetics, metabolism)
- MART-1 Antigen
(chemistry, genetics, metabolism)
- Protein Stability
- Recombinant Fusion Proteins
(chemistry, genetics, isolation & purification, metabolism)
- Reproducibility of Results
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