Aim: 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.
Conclusion: The xenogentic EGFR protein vaccine can overcome the host's immune tolerance problem and induce the production of specific antibodies against.