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目的为研究HPV16 E7蛋白在肿瘤早期预报中的作用及研制治疗性疫苗,特制备HPV16 E7的单克隆抗体。方法将构建成的PGEX4T-1-HPV16 E7的质粒转化BL21细菌,在IPTG诱导下,稳定表达GST-HPV16 E7融合蛋白,免疫Balb/c小鼠,收集免疫小鼠的脾细胞与SP2/0细胞融合,用ELISA双筛,直至检测所有孔均为E7阳性而GST阴性为止,扩大培养、建株、冻存并制备腹水、单克隆抗体。结果成功制备出HPV16 E7的单克隆抗体,可在多种肿瘤组织细胞核中染色。结论该抗体是HPV16 E7蛋白特异性的McAb。
Objective To study the role of HPV16 E7 in early tumor prediction and to develop a therapeutic vaccine to prepare a monoclonal antibody against HPV16 E7. Methods The constructed plasmid pGEX4T-1-HPV16 E7 was transformed into BL21 strain. The GST-HPV16 E7 fusion protein was stably expressed under the induction of IPTG. Balb / c mice were immunized and the spleen cells and SP2 / 0 cells Fusion, double-screen with ELISA until all wells were tested positive for E7 and negative for GST until expanded, cultured, frozen and prepared ascites, monoclonal antibodies. Results The monoclonal antibody of HPV16 E7 was successfully prepared and stained in a variety of tumor cell nuclei. Conclusion The antibody is HPV16 E7 protein-specific McAb.