国产重组抗CD20单克隆抗体在食蟹猴体内的毒代动力学检测方法研究

Research on Toxicokinetics Detection Method of Domestic rh-anti-CD20 Monoclonal Antibody in Cynomolgus Monkey

目的建立一种灵敏度高、特异性好且快速的酶联免疫吸附测定(ELISA)方法,用以检测食蟹猴体内的国产重组抗CD20单克隆抗体(YKY-101)。方法采用双抗夹心的ELISA方法对YKY-101进行定量,以大鼠抗Rituximab为一抗,加入YKY-101后,再加入辣根过氧化物酶(HRP)标记的羊抗人Ig G,加入底物显色,采用酶标仪读取450 nm(参比波长630 nm)处吸光度值。结果建立并验证了灵敏度高、稳定性良好的检测猴血清中YKY-101的ELISA方法。结论该ELISA方法能够满足YKY-101临床前毒代动力学的要求,可应用于猴血清中YKY-101的浓度检测。

Objective To establish a high sentitive, specific and rapid ELISA method for the determination of rh-anti-CD20 monoclonal antibody （YK-101）in cynomolgus monkey. Methods A quantitative sandwich ELISA for assay ofYKY-101 was developed usingrat anti-Rituximab antibody for capturing and goat anti-human IgG-HRP for detecting.Following that,color was developed by the substrate solution and the rection was stopped by stop solution.Then theplate was analyzed with a microplate reader at a wavelength of 450nm/630nm. Results A high sentitive and stablemethod was established for the determination of YK-101 in cynomolgus monkey.Conclusion Itproved that the ELISAmethod was feasible for the dectetion of YK-101 in serum of cynomolgus monkey,which meet the requirements of thepre-clinical guidance fortoxicokinetics.