抗DR5单克隆抗体ELISA检测新方法的建立  

Development of Enzyme Linked Immunosorbent Assay for Quantita tive Determination of Agonistic DR5 Monoclonal Antibody

作　　者：郭聪[1,2,3] 陈知航[2] 许先兴[4] 刘运龙[2] 车津晶[2] 董俊兴[1,3] 程远国[2] 

机构地区：[1]北京工业大学生命科学与生物工程学院,北京100124 [2]军事医学科学院微生物流行病研究所,北京100071 [3]军事医学科学院放射与辐射医学研究所,北京102206 [4]第二炮兵总医院药学部,北京100088

出　　处：《生物技术通讯》2013年第2期209-213,共5页Letters in Biotechnology

基　　金："十二五"国家重大科技专项(2011ZX09102-001-26)

摘　　要：目的：建立一种灵敏、特异、快速的ELISA方法，用于猕猴血清中抗死亡受体5（DR5）单克隆抗体的检测。方法：采用双抗夹心ELlSA法，用猴血清吸附的羊抗入IgG包被于96孔酶标板，加入待测样品，用HRP标记的猴血清吸附的羊抗人IgG进行检测，加底物显色，读取D450值。结果：建立了检测抗DR5单克隆抗体的ELISA方法并进行了确证，方法的线性范围为12．5-800ng／mL，定量下限为12．5ng／mL，板内和板间精密度均小于15％，准确度为-4-15％，冻融稳定性和稀释稳定性良好。结论：方法学确证结果表明，本研究建立的抗DR5单克隆抗体检测方法符合新生物制品临床前药代动力学研究指导原则要求，可用于抗DR5单克隆抗体的检测。Objective： To develop a specific, sensitive and rapid ELISA method for the quantification of agonistic death receptor 5（DRS） monoclonal antibody. Methods： An quantitative sandwich enzyme immunoassay was devel- oped in using goat anti-human IgG for capturing as well as detecting. Following that, color was developed by the substrate solution and the reaction was stopped by stop solution. Finally the plate was read at a wavelength of 450 nm using a microplate reader. Results： An ELISA assay was developed with a wide dynamic range of con- centrations from 12.5 to 800 ng/mL. The lowest quantification of this assay was 12.5 ng/mL, both accuracy of the intra- and inter-assay were less than 15%. Conclusion： The assay is highly sensitive, accurate, specific, and re- producibility over a wide dynamic range of concentrations, which was proven to be a feasible quantitative method for agonistic DR5 monoclonal antibody analysis.

关 键 词：死亡受体5 单克隆抗体 酶联免疫吸附试验法 药代动力学 

分 类 号：R392.1[医药卫生—免疫学]