利用重组人VEGF165制备单克隆抗体的研究

Study of preparing the monoclonal antibody of rhVEGF165

目的 :制备抗重组人血管内皮生长因子 (VEGF) 16 5单克隆抗体。方法 :构建人 VEGF16 5原核表达载体 p ET- 3a/ VEGF16 5 ,并使其在大肠杆菌中高效表达 ,纯化表达产物。用初步纯化的rh VEGF16 5免疫小鼠制成杂交瘤细胞 ,小鼠腹腔接种 ,收集腹水纯化后制成抗 rh VEGF16 5单克隆抗体。结果 :VEGF16 5诱导的内皮细胞增殖可被本研究制备的单克隆抗体中和 ,并表现出较好的剂量依赖关系。结论 :本研究利用 DNA重组技术制备人 VEGF16 5 ,并利用它进一步制备了抗重组人VEGF16 5单克隆抗体。

Objective:To prepare the monoclonal antibody of VEGF165.Methods:The recombinant plasmid pET 3a/VEGF165 was highly expressed in E coli.The mouse was immunized with primarily purified rhVEGF165.Then the hybridized tumor was prepared,and then incubated into the peritoneal cavity of mouse.The monoclonal antibody of rhVEGF165 was then prepared after the purification of the gathered ascites. Results: Proliferation of endothelial cell induced by VEGF165 was dose dependently neutralized by the monoclonal antibody prepared in our study.Conclusion:The recombinant human VEGF165 was expressed in E coli . And then the monoclonal antibody of rhVEGF165 was prepared with the expressed protein.