發(fā)布時間：2019-06-27 07:36

【摘要】：目的建立一種高特異、高靈敏、新型的定量檢測食蟹猴血清中PD-1單克隆抗體質(zhì)量濃度的ELISA方法。方法用重組人PD-1作為抗原包被到96孔板上,加入系列濃度標準品(PD-1單克隆抗體)和稀釋的食蟹猴血清,再加入稀釋的猴血清吸附的羊抗人Ig G-HRP(二抗),二抗與結(jié)合到抗原上的PD-1單克隆抗體特異性結(jié)合,加入底物顯色劑,用硫酸終止,在450 nm下讀數(shù)。結(jié)果在40~0.625 ng/ml范圍內(nèi),PD-1單克隆抗體質(zhì)量濃度的對數(shù)值與吸光值呈S形曲線,方法靈敏度為0.625 ng/ml,TNF-α、r HSA、PEG-GH和受試品PD-1單克隆抗體均無交叉反應,方法的回收率為94.5%~98.0%,板內(nèi)精密度波動在-4.1%~-0.1%,板間精密度波動在-5.0%~-1.0%。結(jié)論本方法符合新生物制品臨床前藥代動力學研究指導原則的要求,可用于食蟹猴體內(nèi)PD-1單克隆抗體的定量檢測。
[Abstract]:Objective to establish a highly specific, highly sensitive and novel ELISA method for quantitative determination of PD-1 monoclonal antibody in serum of cynomolgus monkeys. Methods Recombinant human PD-1 was used as antigen coated on 96 well plate, a series of concentration standard (PD-1 monoclonal antibody) and diluted cynomolgus monkey serum were added, and then sheep anti-human Ig G-HRP (second antibody) adsorbed by diluted monkey serum was added. The second antibody specifically binds to the PD-1 monoclonal antibody bound to the antigen. The substrate developer is added, terminated with sulfuric acid and read at 450 nm. Results in the range of 40 鈮，

本文編號：2506611