抗KDR3单克隆抗体的制备和生物活性的鉴定

Generation and Characterization of Monoclonal Antibodies Against Domain 3 of VEGF Receptor II

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 the monoclonal antibody directing against KDR3of VEGF receptor II and to study its biological activity.Methods:The ascitic fluid was purified by protein G,and its titer was assayed by ELASA.The memberane of ECV304was prepared to de-tected the binding activity of the monoclonal antibody.Results:A monoclonal antibody was ob-tained,which could inhibit proliferation of ECV304induced by VEGF.Conclusion:The antibody directing against KDR3inhibited proliferation of ECV304by blocking VEGF binding to KDR and could be potent in the treatment of tumor.