【摘要】 目的:建立一种新型、快速定量检测食蟹猴血清中抗血管内皮生长因子受体2（VEGFR2）单克隆抗体浓度的方法。方法:将特异性抗原VEGFR2-His包被在固相载体上,加稀释的受试药品血清,然后加入HRP标记的羊抗人IgG-（h+1）,再加入TMB显色液,最后加入1 mol/L硫酸终止液,在酶标仪上用双波长读取D_450/560nm值。结果:建立了定量检测食蟹猴血清中抗VEGFR2单克隆抗体浓度的ELISA法,方法的线性范围为400⁶.25 ng/m L,定量下限为6.25ng/m L,板内精密度介于-13.6%⁸.3%,板间精密度介于-6.1%⁶.3%,与Avastin、Actemra、Cetuximab均无交叉,室温稳定性及冻融稳定性良好,无稀释效应。结论:通过方法学的确证,本实验建立的方法可满足抗VEGFR2单克隆抗体在食蟹猴体内的药代动力学研究要求,可用于抗VEGFR2单克隆抗体的检测。更多还原

【Abstract】 Objective: To establish a new and rapid ELISA method for quantitive determination of anti-vascularendothelial growth factor receptor 2(VEGFR2) monoclonal antibody in the blood serum of Macaca fascicularis.Methods: The specific antigen VEGFR2- His bag was coated on the solid phase carrier, and diluted serum wasadded, followed by goat anti- human Ig G- HRP as detecting antibody, TMB color liquid and sulfuric acid as termi-nation liquid. Finally, the absorbance of each well was measured by a microplate reader under 450/560 nm. Results: The linearity range of the new ELISA method was 400~6.25 ng/m L, the lower limit of quantification was6.25 ng/m L, the within- plate precision ranged from- 13.6% to 8.3%, the between- plates precision was- 6.1% to 6.3%. There was no cross-reaction with Avastin, Actemra, and Cetuximab. Room temperature and freeze-thaw sta-bility were validated, and no dilution effet was found. Conclusion: This ELISA method met the requirements ofpharmacokinetic study, and could be used to determine anti-VEGFR2 monoclonal antibody in cynomolgus monkeys.更多还原