【摘要】 目的测定人源化抗人血小板膜糖蛋白Ⅲa(GPⅢa)单克隆抗体813F(ab)2片段[813-F(ab)2]与人、猕猴、毕格犬血小板结合的亲和常数。方法将人、猕猴和毕格犬的血小板分别包板,生物素标记813-F(ab)2,竞争性ELISA法测定结合到包被血小板上的813-F(ab)2的结合量,Scatchard分析计算出813-F(ab)2与不同种属血小板结合的亲和常数。结果813-F(ab)2与人、猕猴和毕格犬的血小板结合的亲和常数分别为(3.21±3.04)×108L/mol、(1.51±1.39)×107L/mol和(3.62±2.16)×107L/mol,其亲和力由大到小依次为人、毕格犬、猕猴。结论813-F(ab)2与人、猕猴和毕格犬的血小板均有较强的特异性结合能力,为813-F(ab)2的后续药效学评价、毒理分析等提供了基础。更多还原

【Abstract】 Objective To assess affinity constant of humanized monoclonal antibody fragment 813 [813-F(ab)2] against human platelet membrane glycoproteinⅢa(GPⅢa)binding to platelets of different category animals.Methods 813-F(ab)2 was labeled with biotin.The platelets of human,macaque and dog were coated respectively.The binding quantity of 813-F(ab)2 with coated platelets was measured with competive ELISA method and the affinity constant of 813-F(ab)2 antibody binding to platelets of human,macaque and dog was calculated with scatchard analysis.Results The affinity constant of humanized antibody 813-F(ab)2 binding to platelets of human,macaque and dog was(3.21±3.04)×108 L/mol,(1.51±1.39)×107 L/mol,(3.62±2.16)×107 L/mol.Conclusion The humanized antibody 813-F(ab)2 can strongly bind to the platelets of human,macaque and dog.Thus,it will provide a basis for its pharmacodynamics estimation and toxicological analysis更多还原