用于测量生物体液中前列腺素E2的高特异性和高灵敏度免疫测定法。
Highly specific and sensitive immunoassay for the measurement of prostaglandin E2 in biological fluids.
作者信息
Gao Yuan, Hou Rui, Liu Fen, Cai Runlan, Fang Lei, Peng Cheng, Qi Yun
机构信息
Institute of Medicinal Plant Development, Chinese Academy of Medical Sciences & Peking Union Medical College, Beijing 100193, China.
Pharmacy College, Chengdu University of Traditional Chinese Medicine, Chengdu 610075, China.
出版信息
Bioanalysis. 2015 Oct;7(19):2597-607. doi: 10.4155/bio.15.130. Epub 2015 Oct 12.
BACKGROUND
Lack of specificity of anti-PGE2 antibodies is a long-standing problem. Given quite a few analogs and low PGE2 content in biological fluids, it is quite important to simultaneously meet the demands of high specificity and sensitivity.
RESULTS
Highly specific anti-PGE2 antibodies were obtained by combined use of cationic carrier protein and Mannich reaction. The cross-reactivity values of the resultant polyclonal and monoclonal antibodies against eight analogs were <14 and <5%, respectively. Furthermore, we established a highly sensitive ELISA, which could be applied to direct analysis of PGE2 at the pg/ml level (LOQ = 15.6 pg/ml).
CONCLUSION
We provide an appropriate strategy to develop a highly-specific and sensitive immunoassay for measuring low PGE2 content in biological samples.
背景
抗前列腺素E2（PGE2）抗体缺乏特异性是一个长期存在的问题。鉴于生物体液中有不少类似物且PGE2含量较低，同时满足高特异性和高灵敏度的要求非常重要。
结果
通过阳离子载体蛋白与曼尼希反应联合使用获得了高度特异性的抗PGE2抗体。所得多克隆抗体和单克隆抗体对八种类似物的交叉反应值分别<14%和<5%。此外，我们建立了一种高灵敏度的酶联免疫吸附测定法（ELISA），可用于直接分析pg/ml水平的PGE2（定量下限=15.6 pg/ml）。
结论
我们提供了一种合适的策略，用于开发一种高特异性和高灵敏度的免疫测定法，以测量生物样品中低含量的PGE2。
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