定量检测重组抗白介素6受体人源化单克隆抗体HS628的ELISA方法的建立及确证  

Development of an Enzyme Linked Immunosorbent Assay for Quantitative Measurement of HS628,an Anti-IL-6 Receptor Antibody

作　　者：张代超 魏峰[1] 毛素梅 刘运龙[3] 任欣怡[3] 李金龙[1] 陈知航[3] 程远国[3] 

机构地区：[1]吉林农业大学生命科学学院,吉林长春130062 [2]广西桂东灵长类开发实验有限公司,广西桂东543000 [3]军事医学科学院微生物与流行病研究所,北京100071

出　　处：《生物技术通讯》2015年第5期672-677,共6页Letters in Biotechnology

摘　　要：目的：建立一种灵敏、特异、快速、经济的ELISA方法,用于检测食蟹猴血清中重组抗白介素6受体人源化单克隆抗体HS628的含量。方法：对ELISA与生物素-亲和素ELISA（BA-ELISA）方法进行比较,最终采用BA-ELISA法对HS628进行定量。包被抗体为羊抗人Ig G单抗,以生物素标记的重组白介素6受体作为检测抗原,以HRP标记的链霉亲和素作为放大剂,最终用单组分显色液（TMB）显色。结果：建立了特异性检测HS628的ELISA方法并完成方法学确证,该方法的线性范围为1.64~400 ng/m L,灵敏度为1.64 ng/m L,精密度、准确度、回收率及稳定性符合要求,与对照品校正标准曲线吻合。结论：用建立的ELISA方法测定猴血清中HS628的含量能满足新生物制药临床前药代动力学研究指导原则要求,可用于HS628的检测。Objective： To develop a sensitive, specific, rapid and economic ELISA method for detecting the con- centration of HS628, a recombinant anti-IL-6 receptor antibody, in cynomolgus monkey serum. Methods： Compar- ing with federation ELISA, a bridged avidin biotin ELISA for HS628 measurement was developed, using goat anti- human IgG monoclonal antibody for capturing, bion-rhlL-6Rα for detecting and streptavidin-HRP for signal ampli- fication. After all, color was developed by the substrate solution. Results： A special detection method of ELISA was developed, with a wide dynamic range of concentrations from 1.64 to 400 ng/mL, and with a lowest quantifica- tion of 1.64 ng/mL. Meanwhile, the accuracy, precision, recovery rate and stability all could meet the require- ments. Conclusion： This method is highly sensitive, accurate, specific and reproducibility, which was proven to be a feasible quazltitative method for the detection of HS628.

关 键 词：重组抗白介素6受体人源化单克隆抗体 间接ELISA法 生物素-亲和素ELISA法 药代动力学 

分 类 号：R392.1[医药卫生—免疫学] R969.1[医药卫生—基础医学]