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目的:建立检测食蟹猴血浆中TNF-α单克隆抗体的间接ELISA检测方法。方法:利用包被抗原TNF-α及酶标抗体山羊抗人IgG-HRP构建TNF-α单克隆抗体的ELISA检测试剂盒,并进行方法学验证。结果:间接ELISA法检测抗TNF-α抗体的最低检出限为0.2 ng·mL-1;最低定量限为0.5 ng·mL-1;浓度0.5~200 ng·mL-1范围内线性良好。日内、日间回收率分别为85.5%~95.0%和85.3%~94.0%;日内、日间精密度分别为4.6%~15.0%和1.1%~8.0%。结论:本实验成功建立了TNF-α单克隆抗体的间接ELISA检测方法,可用于食蟹猴血浆中TNF-α单克隆抗体的含量测定。
Objective: To establish an indirect ELISA method for the detection of monoclonal antibody against TNF-α in plasma of cynomolgus monkeys. Methods: The ELISA kit for detecting TNF-α monoclonal antibody was constructed by using coated antigen TNF-α and enzyme-labeled goat anti-human IgG-HRP, and the method was validated by the method. Results: The minimum detectable limit of detection of anti-TNF-α antibody by indirect ELISA was 0.2 ng · mL-1. The lowest limit of quantification was 0.5 ng · mL-1. The linearity was good in the range of 0.5 ~ 200 ng · mL-1. The daily and daily recoveries were 85.5% -95.0% and 85.3% -94.0%, respectively. The intra-day and inter-day recoveries were 4.6% -15.0% and 1.1% -8.0%, respectively. Conclusion: The indirect ELISA method of TNF-α monoclonal antibody was successfully established in this experiment and could be used to determine the content of monoclonal antibody of TNF-α in plasma of cynomolgus monkeys.