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Expression and purification of the cancer antigen SSX2: A potential cancer vaccine
- Source :
-
Protein Expression & Purification . Dec2007, Vol. 56 Issue 2, p212-219. 8p. - Publication Year :
- 2007
-
Abstract
- Abstract: SSX2 is a cancer testis antigen expressed in a wide variety of cancers, including synovial sarcoma and melanoma. It holds promise as a potential antigen for cancer immunotherapy. A process for the production of recombinant SSX2 was developed by overexpressing a His-tagged fusion protein of SSX2 in Escherichia coli C41 (DE3). A T-7 promoter system was employed and a plasmid was introduced into the strain to compensate for rare codons in the SSX2 sequence. The production of SSX2 was scaled up to a 2-L fermentation that was operated under fed-batch conditions to improve productivity. After 32h cultivation, the wet cell mass reached 260mg/ml, with SSX2 produced mainly as inclusion bodies at a concentration of 1.1g/L. Urea-solubilized SSX2 was purified by nickel affinity, ion exchange and hydrophobic interaction chromatography. The recovery of SSX2 was 20%, and over 87% purity was obtained with an endotoxin level of 0.11EU/μg. The purified recombinant SSX2 was characterized by ELISA and was shown to be recognized by human sera that have been reported to carry anti-SSX2 antibodies. [Copyright &y& Elsevier]
- Subjects :
- *CANCER treatment
*ENDOCRINE glands
*NEUROENDOCRINE tumors
*ESCHERICHIA coli
Subjects
Details
- Language :
- English
- ISSN :
- 10465928
- Volume :
- 56
- Issue :
- 2
- Database :
- Academic Search Index
- Journal :
- Protein Expression & Purification
- Publication Type :
- Academic Journal
- Accession number :
- 27227959
- Full Text :
- https://doi.org/10.1016/j.pep.2007.07.009