人血清apo M的ELISA检测方法评价

Methodological evaluation of sandwich ELISA for detecting concentration of apo M in human plasma

目的建立并评价检测人血清载脂蛋白M(apo M)浓度的双抗体夹心ELISA法。方法用抗apo M单克隆抗体作包被抗体,辣根过氧化物酶(HRP)标记的抗apo M单克隆抗体作酶标记抗体,建立检测apo M的双抗体夹心ELISA法。检测483例体检健康者的血清apo M浓度,建立本实验室的参考值。结果线性范围为31.3～500 ng/mL,最低检测限为23.9 ng/mL;脂蛋白(a)、apo A1和apo B对apo M检测结果干扰作用小;批内、批间变异系数(CV)均<10%;参考值为(22.1±12.1)μg/mL。结论建立检测人血清apo M浓度的双抗体夹心ELISA法,为研究apo M的生理功能及其在脂质代谢中的作用提供了一种新的研究工具。

Objective To establish an ELISA to detect the concentration of apolipoprotein M（apo M） in human plasma,and evaluate the methodological features.Method A sandwich ELISA was developed by using anti-apo M monoclonal antibody as the coated antibody,and horseradish peroxidase（HRP） labeled anti-apo M monoclonal antibody as the enzyme-labeled antibody.The developed method was used to investigate normal apo M variation and establish the reference value in this laboratory by measuring 483 samples of healthy individuals.Result The linear range of the method presented a linear range of 31.3-500 ng/mL with a lowest detectable concentration of 23.9 ng/mL.The interferences of Lp（a）,apo AI and apo B to apo M detection were minor.The intra-assay coefficient of variation（CV） and inter-assay CV of the ELISA were both less than 10%.The overall reference interval of apo M was（22.1±12.1） μg/mL.Conclusions A sandwich ELISA to measure the apo M concentration in human plasma was developed,and the method could serve as a useful tool for the research on apo M function to understand its role in lipid metabolism.