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目的:制备抗KDR3单克隆抗体,研究其阻断VEGF受体的作用。方法:将抗KDR3单克隆抗体腹水,采用ELISA酶免疫分析方法,测定其效价,提取ECV304细胞膜蛋白用Westernblot方法进行识别抗原的鉴定,并测定抗KDR3单克隆抗体的活性。结果:获得一株稳定分泌抗KDR3的杂交瘤细胞株,所产生抗体可阻断VEGF对人脐静脉内皮细胞系ECV304的刺激增生作用。结论:抗KDR中和抗体IMC-ICI和抗KDR3单克隆抗体同样对ECV304细胞都表现出对外源性VEGF有明显抑制作用,而且对内源性VEGF也有明显抑制作用。抗KDR3单克隆抗体在肿瘤治疗的过程中将起一定的作用。
OBJECTIVE: To prepare anti-KDR3 monoclonal antibody to study its function of blocking VEGF receptor. Methods: The anti-KDR3 monoclonal antibody ascites was measured by enzyme-linked immunosorbent assay (ELISA), the titer was determined and the protein of ECV304 was extracted and identified by Western blot. The anti-KDR3 monoclonal antibody activity was determined. Results: A hybridoma cell line stably secreting anti-KDR3 was obtained and the antibody produced could block the stimulatory effect of VEGF on human umbilical vein endothelial cell line ECV304. CONCLUSION: Anti-KDR neutralizing antibody IMC-ICI and anti-KDR3 monoclonal antibody also showed significant inhibitory effect on exogenous VEGF in ECV304 cells as well as significant inhibitory effect on endogenous VEGF. Anti-KDR3 monoclonal antibody in the treatment of cancer will play a role.