食蟹猴血浆中TNF-α单克隆抗体间接ELISA检测法的建立
Establishment of indirect ELISA for detecting TNF-α monoclonal antibody in plasma of machin
分类号:
出版年·卷·期(页码):2014,34 (9):0-0
DOI:
10.16155/j.0254-1793.2017.01.01
-----摘要:-------------------------------------------------------------------------------------------
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。
-----英文摘要:---------------------------------------------------------------------------------------
Objective: To establish an indirect ELISA detection method for TNF-α monoclonal antibody in plasma of machin. Methods: An indirect ELISA kit was built by utilizing the anti-TNF-α monoclonal antibody and goat anti-human IgG-HRP,which was used as a coating antibody and a detecting antibody respectively.Methodology validations were performed for the method. Results: The detection limit for TNF-α was 0.2 ng·mL-1;the limit of quantification was 0.5 ng·mL-1;the standard curve of established ELISA was linear between 0.5 and 200 ng·mL-1.Inter-day accuracy and intra-day accuracy were 85.5%-95.0%,85.3%-94.0%,respectively.Inter-day precision and intra-day precision were 4.6%-15.0%,1.1%-8.0%,respectively. Conclusion: An indirect ELISA is developed,providing a useful tool for quantifying the level of TNF-α in plasma of machin.
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