定量检测PD-1单克隆抗体ELISA法的建立

发布时间：2019-06-27 07:36

【摘要】：目的建立一种高特异、高灵敏、新型的定量检测食蟹猴血清中PD-1单克隆抗体质量浓度的ELISA方法。方法用重组人PD-1作为抗原包被到96孔板上,加入系列浓度标准品(PD-1单克隆抗体)和稀释的食蟹猴血清,再加入稀释的猴血清吸附的羊抗人Ig G-HRP(二抗),二抗与结合到抗原上的PD-1单克隆抗体特异性结合,加入底物显色剂,用硫酸终止,在450 nm下读数。结果在40~0.625 ng/ml范围内,PD-1单克隆抗体质量浓度的对数值与吸光值呈S形曲线,方法灵敏度为0.625 ng/ml,TNF-α、r HSA、PEG-GH和受试品PD-1单克隆抗体均无交叉反应,方法的回收率为94.5%~98.0%,板内精密度波动在-4.1%~-0.1%,板间精密度波动在-5.0%~-1.0%。结论本方法符合新生物制品临床前药代动力学研究指导原则的要求,可用于食蟹猴体内PD-1单克隆抗体的定量检测。
[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，

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