Expression and purification of the cancer antigen SSX2: a potential cancer vaccine. Academic Article uri icon

Overview

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/microg. 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.

publication date

  • August 24, 2007

Research

keywords

  • Antigens, Neoplasm
  • Neoplasm Proteins
  • Repressor Proteins

Identity

Scopus Document Identifier

  • 35448983175

Digital Object Identifier (DOI)

  • 10.1016/j.pep.2007.07.009

PubMed ID

  • 17931884

Additional Document Info

volume

  • 56

issue

  • 2