人乳头瘤病毒16型E7蛋白单克隆抗体的研制

Preparation of a monoclonal antibody against the human papillomavirus type 16 E7(HPV 16 E7) protein

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 An HPV 16 E7 monoclonal antibody was prepared.This antibody will be useful for diagnosing early cancers and developing therapeutic vaccines.Method A plasmid constructed with PGEX4T-1-HPV16 E7 was transformed into E.coli BL21,and expression of the fusion protein was induced by IPTG.Spleen cells from immunized Balb/c mice were fused with SP2/0 cells by a routine method,and the valence of the antibody was evaluated using ELISA.Expansion,isolation and freezing of clones were carried out,and finally an ascitic fluid which contained monoclonal antibodies was produced.Result The HPV16 E7 monoclonal antibody was found specifically in the nuclei of a variety of cancers tissues.Conclusion The HPV16 E7 protein antibody is a specific McAb.