[Preparation of human nasopharyngeal carcinoma bivalent anti-idiotype antibody vaccine and identification of its activity].
OBJECTIVE
To construct a prokaryotic expression vector expressing a fully human bivalent anti-idiotype antibody and identify the expression product.


METHODS
The genes of G22, I50 were subcloned into prokaryotic expression vector pET25b (+) in order and confirmed by colony PCR, restriction enzyme digestion, DNA sequencing.The positive construct was transformed into E.coli BL21 (DE3) and the protein was expressed after induction by IPTG. The identity of the expressed protein was confirmed by Western blot. The activity of the protein was analysed by ELISA and further by lymphocyte proliferation assay.


RESULTS
The recombinant vector was constructed successfully, the recombinant protein was successfully expressed and purified with a purification of about 90%. The molecular weight of the expressed protein was 42000 which was in accordance with expectation. The activity of expressed protein after renaturation had recovered and further confirmed by lymphocyte proliferation assay in vitro.


CONCLUSIONS
We obtained an active bivalent anti-idiotype antibody which laid a foundation for the clinical immunotherapy of nasopharyngeal carcinoma.
Wang
Jia-jia
JJ
Laboratory of Tumor Immunobiology, Cancer Research Institute, Central South University, Changsha 410078, China. xywjj08@163.com
Guo
Feng-jie
FJ
Li
Yue-hui
YH
Li
Guan-cheng
GC
D000888:Antibodies, Anti-Idiotypic / Q000235:genetics / Q000276:immunology*; D000951:Antigens, Neoplasm / Q000235:genetics / Q000276:immunology; D019496:Cancer Vaccines / Q000235:genetics / Q000276:immunology*; D006801:Humans; D009303:Nasopharyngeal Neoplasms / Q000276:immunology* / Q000517:prevention & control; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000276:immunology
D000888:Antibodies, Anti-Idiotypic; D000951:Antigens, Neoplasm; D019496:Cancer Vaccines; D011993:Recombinant Fusion Proteins