重组人源化兔抗血管内皮生长因子单克隆抗体的抗原结合活性测定  

Determination of Antigen Binding Activity of Recombinant Humanized Rabbit Anti-Vascular Endothelial Growth Factor Monoclonal Antibody

作　　者：范文红[1] 毕华[1] 饶春明[1] 

机构地区：[1]中国食品药品检定研究院重组技术产品室卫生部生物技术产品检定方法及其标准化重点实验室,北京100050

出　　处：《中国生物制品学杂志》2011年第9期1079-1080,1086,共3页Chinese Journal of Biologicals

基　　金：国家科技重大专项课题(2009ZX09307-001)

摘　　要：目的采用直接ELISA法测定重组人源化兔抗血管内皮生长因子(Vascular endothelial growth factor,VEGF)单克隆抗体的抗原结合活性。方法以VEGF-Fc为包被抗原,采用直接ELISA法测定不同浓度重组人源化兔抗vEGF单抗参考品和供试品与抗原的结合活性,根据供试品及参考品的EC50值计算供试品的相对结合力,并分析该法的精密性。结果重组人源化兔抗VEGF单抗供试品及参考品的抗原结合均存在量效关系,且符合四参数方程。同一批次的供试品单抗原液的相对结合活性分别为参考品的(90.43±18.74)%、(123.56±18.40)%和(106.83±18.47)%,均值为(106.94±16.57)%。3次试验的变异系数分别为20.72%、14.89%和17.29%。同一批次的供试品成品的相对结合活性分别为(97.34±18.42)%、(123.30±11.08)%和(93.91±4.57)%,变异系数分别为18.93%、8.96%和4.87%。结论 直接ELISA法精密性良好,操作简便,可用于重组人源化兔抗VEGF单抗抗原结合活性的常规检测。Objective To determine the antigen binding activity of recombinant humanized rabbit anti-vascular endothelial growth factor（VEGF） monoclonal antibody（McAb） by direct ELISA.Methods The antigen binding activities of reference and test samples of recombinant humanized rabbit anti-VEGF McAb at various concentrations were determined by direct ELISA using VEGF-Fc as coating antigen,based on which the relative binding activities of test samples were calculated according to the EC50 values of test samples and reference,and the method was analyzed for precision.Results Both the antigen binding activities of reference and test samples of rabbit anti-VEGF McAb showed dose-response relationships and were fitted with four-parameter equation.The relative binding activities of three test samples of bulk McAbs were（90.43 ± 18.74）%,（123.56 ± 18.40）% and（106.83 ± 18.47）%,with a mean of（106.94 ± 16.57）% of those of reference of the same batches,respectively.The variation coefficients of three tests were 20.72%,14.89% and 17.29% respectively.The relative binding activities of three test samples of final products of the same batch were（97.34 ± 18.42）%,（123.30 ± 11.08）% and（93.91 ± 4.57）%,with variation coefficients of 18.93%,8.96% and 4.87%,respectively.Conclusion Direct ELISA showed high precision and was easy to handle,which might be used for routine test for antigen binding activity of recombinant humanized rabbit anti-VEGF McAb.

关 键 词：血管内皮生长因子类 抗体 单克隆 抗原结合活性 酶联免疫吸附测定 

分 类 号：R392-33[医药卫生—免疫学]