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目的:构建异种EGFR蛋白疫苗并研究其抗肿瘤效果。方法:用PCR方法获得鸡EGFR胞外片段,与pGEX-4T-2载体连接,并在E.coli BL21(DE3)中表达鸡的EGFR胞外片段与GST的融合蛋白,融合蛋白经Ni2+亲和层析柱纯化、复性后,免疫小鼠,免疫3次后,接种小鼠Lewis细胞,观测肿瘤生长情况,ELISA检测小鼠血清中抗EGFR蛋白的抗体效价。结果:成功构建了EGFR胞外段基因的原核表达载体;SDS-PAGE显示成功表达了目的融合蛋白;融合蛋白免疫小鼠后,ELISA法检测到特异性抗EGFR抗体;实验组与对照组肿瘤体积的比较结果显示,融合蛋白疫苗能够在一定程度上抑制肿瘤的生长。结论:异种EGFR蛋白能够克服免疫耐受问题,诱导机体产生抗体,有一定的抗肿瘤效果,为后续的肿瘤疫苗研究打下了基础。
Objective: To construct different EGFR vaccine and study its anti-tumor effect. Methods: The extracellular fragment of chicken EGFR was obtained by PCR and ligated with pGEX-4T-2 vector. The fusion protein of EGFR extracellular fragment and GST was expressed in E. coli BL21 (DE3) Purification of the column, renaturation, immunization of mice, immunized three times after inoculation of Lewis cells in mice, observation of tumor growth, ELISA detection of serum anti-EGFR antibody titers. Results: The prokaryotic expression vector of EGFR extracellular domain gene was successfully constructed. The fusion protein was successfully expressed by SDS-PAGE. The specific anti-EGFR antibody was detected by ELISA. The tumor volume of experimental group and control group The results of the comparison show that the fusion protein vaccine can inhibit tumor growth to a certain extent. CONCLUSION: The heterologous EGFR protein can overcome the immune tolerance problem, induce the body to produce antibody and have certain antitumor effect, which lays the foundation for the subsequent study of tumor vaccine.