抗人VEGF受体KDR胞外区域3血清的制备及生物活性

Preparation and Its Biological Activity of Serum Directing against Domain 3 of VEGF Receptor II

目的:制备抗人VEGF受体KDR胞外区域3(KDR3)的血清,研究其阻断VEGF受体的作用。方法:采用6周龄雌性BALB/C小鼠将纯化的人VEGF受体II基因3区蛋白质与完全佐剂和不完全佐剂制成抗原乳剂,分3次注射,前2次为腹腔注射,第3次为静脉注射,待第3次静脉注射后7天摘眼球取血,静置凝固后取上层血清测抗血清的效价,用血管内皮细胞ECV3042×104/ml铺24孔板,加入VEGF的同时加入抗KDR中和抗体IMC-ICI和KDR3抗血清,从次日开始测定。结果:用Studentt-tes统计学方法处理显示,PBS对照组与非相关性血清组P>0.05,抗KDR中和抗体IMC-ICI组与KDR3抗血清组P>0.05,而VEGF组与非相关性血清组和KDR3抗血清组P<0.05。结论:VEGF对ECV304细胞有促增殖作用,而抗KDR中和抗体IMC-ICI和KDR3抗血清对ECV304细胞都表现出了对外源性VEGF有明显抑制作用,同时对内源性VEGF也有明显抑制作用。

O bjective:To prepare the anti-serum directing against KDR3of VEGF recep-tor I I and to study its biological activity.Methods:The anti-serum directing again st KDR3was prepared from mice immunized with a soluble form of KDR3.Its titer was assayed by ELISA.Its activities were detected.Results:Student ,s t-tes t indicated that treatment with VEGF group and non-relative serum group,VEGF group and KDR3anti-serum group were significantly different (P<0.05).C onclusion:VEGF induced proliferation of ECV304.The anti-serum directing ag ainst KDR3inhibited proliferation of ECV304by blocking VEGF binding to KDR.