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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。
OBJECTIVE: To prepare a serum of anti-human VEGF receptor KDR extracellular domain 3 (KDR3) and study its effect on blocking VEGF receptor. Methods: The 6-week-old female BALB / C mice were used to make the purified antigen of human VEGF receptor region 3 protein with complete adjuvant and incomplete adjuvant, The third for intravenous injection, to be taken after the third intravenous injection of the ball 7 days after taking the blood, after standing coagulation serum antiserum taken upper titers with vascular endothelial cells ECV3042 × 104 / ml 24-well plate, add VEGF plus anti-KDR neutralizing antibodies IMC-ICI and KDR3 antisera were measured starting the next day. Results: Student’s t-test showed that P> 0.05 in PBS control group and non-related serum group, P> 0.05 in anti-KDR neutralizing antibody IMC-ICI group and KDR3 antiserum group, but not in VEGF group Serum group and KDR3 antiserum group P <0.05. CONCLUSION: VEGF can promote the proliferation of ECV304 cells, while the anti-KDR neutralizing antibodies IMC-ICI and KDR3 antiserum have obvious inhibitory effects on exogenous VEGF and ECV304 cells as well as endogenous VEGF .