title: Expression and activity identification of a human nasopharyngeal carcinoma I50 anti-idiotype antibody.
abstract: OBJECTIVE
  To obtain I50 anti-idiotype antibody and identify its activity in vitro.
  
  
  METHODS
  I50 anti-idiotype (Id) antibody gene was amplified from the template of fuse 5-I50 by PCR to construct a prokaryotic expression vector pET25b-I50. The expression of pET25b-I50 in E. coli BL21(DE3) was induced by isopropylthio-β-D-galactopyranoside (IPTG) and was confirmed by SDS-PAGE and Western blot with Ab1(FC2) monoclonal antibody and an anti-hexahistidine tag antibody. The method of dialysis refolding was used to restore the activity of I50 anti-Id antibody, which was measured by Dot-ELISA and lymphocyte proliferation assay.
  
  
  RESULTS
  The recombinant vector was successfully constructed and the recombinant protein was successfully expressed and purified with 90% purity. The relative molecular weight of the expressed protein was 15 kD, which was in accordance with expectation. The activity of I50 anti-Id antibody could be restored and could promote the proliferation of lymphocyte in a dose-dependent manner.
  
  
  CONCLUSIONS
  These results suggested that I50 anti-Id protein vaccine is likely an option in the therapy against nasopharyngeal carcinoma in vivo.
authors.affiliation: Cancer Research Institution, Central South University, Changsha, China.
authors.forename: Jiajia
authors.initials: J
authors.lastname: Wang
authors.forename: Yalin
authors.initials: Y
authors.lastname: Li
authors.forename: Fengjie
authors.initials: F
authors.lastname: Guo
authors.forename: Guohua
authors.initials: G
authors.lastname: Zhou
authors.forename: Guancheng
authors.initials: G
authors.lastname: Li
mesh_terms: D000888:Antibodies, Anti-Idiotypic / Q000096:biosynthesis* / Q000235:genetics / Q000276:immunology*; D000951:Antigens, Neoplasm / Q000235:genetics / Q000276:immunology*; D019496:Cancer Vaccines / Q000235:genetics / Q000276:immunology; D045744:Cell Line, Tumor; D004926:Escherichia coli / Q000235:genetics / Q000378:metabolism; D005822:Genetic Vectors / Q000235:genetics; D006801:Humans; D009303:Nasopharyngeal Neoplasms / Q000276:immunology* / Q000473:pathology; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000235:genetics / Q000276:immunology
chemical_list: D000888:Antibodies, Anti-Idiotypic; D000951:Antigens, Neoplasm; D019496:Cancer Vaccines; D011994:Recombinant Proteins