[Expression of xenogentic epidermal growth factor receptor ectodomain and its antitumor effect].
OBJECTIVE
To construct the EGFR protein vaccine and evaluate its antitumor effect.


METHODS
The gene of chicken EGFR ectodomain was amplified by PCR, inserted into the pGEX-4T-2 vetor, and then exporessed in E.coli BL21 with glutathione S-transferase in a fusion protein form. The fusion protein was purified by metal affinity chromatography, and refolded by dialysis.Then the mice were immunized with the fusion protein three times. After that, these mice were vaccinated with Lewis cells. At last the growth of tumors was measured and the surm antibody response was measured by ELISA.


RESULTS
The ectodomain gene of EGFR was ligated into prokaryotic expression vetor. The expression of the fusion protein was analyzed by SDS-PAGE. The relative moleclar mass of the protein is about M(r); 50,000. After the third immunization, all the mice immunized with the fusion protein showed an antibody response towards the EGFR protein. Compared with the control group, the fusion protein group showed good anti-tumor effects.


CONCLUSIONS
The xenogentic EGFR protein vaccine can overcome the host's immune tolerance problem and induce the production of specific antibodies against.
Li
Ying-hui
YH
College of Animal Husbandry and Veterinary Science, Northwestern Sci-Tech University of Agriculture and Forestry, Yangling, 712100, China.
Tang
Liang
L
Liu
Qian
Q
Chen
De-kun
DK
Tan
Li-song
LS
D000818:Animals; D019496:Cancer Vaccines / Q000235:genetics / Q000276:immunology; D045744:Cell Line, Tumor; D002645:Chickens; D002846:Chromatography, Affinity; D004195:Disease Models, Animal; D004591:Electrophoresis, Polyacrylamide Gel; D004797:Enzyme-Linked Immunosorbent Assay; D066246:ErbB Receptors / Q000235:genetics / Q000276:immunology* / Q000378:metabolism*; D005260:Female; D005982:Glutathione Transferase / Q000235:genetics / Q000378:metabolism; D006801:Humans; D008175:Lung Neoplasms / Q000276:immunology; D051379:Mice; D008810:Mice, Inbred C57BL; D016133:Polymerase Chain Reaction; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000276:immunology* / Q000378:metabolism*
D019496:Cancer Vaccines; D011993:Recombinant Fusion Proteins; D005982:Glutathione Transferase; D066246:ErbB Receptors