检测人IL 37双抗夹心ELISA方法的建立

目的：建立定量检测人血清IL-37的双抗夹心ELISA。方法：以鼠抗人IL-37单抗作为捕获抗体，制备的兔抗人IL-37多抗作为检测抗体，HRP标记山羊抗兔IgG为二抗，重组人IL-37蛋白为标准品，建立检测人IL-37的双抗夹心ELISA方法。并对该方法的工作条件进行优化，对其灵敏度、线性范围、重复性和对登革热非结构蛋白NS1阳性患者血清IL-37的检测效果进行评价。结果：重组IL-37蛋白为标准品建立的双抗夹心ELISA法检测灵敏度为1.465 μg/L，线性范围为1.465~46.875 μg/L，批内和批间变异系数分别为6.6%和11.7%。采用此方法对诊断为登革热的患者血清进行检测，结果显示非结构蛋白NS1阳性患者IL-37水平显著高于健康人对照组。结论：成功建立了双抗夹心ELISA检测方法，可用于人血清中IL-37的检测。

Establishment of a double antibody sandwich ELISA for detection of human IL-37

Objective:To establish a double antibody sandwich ELISA assay for detection of human IL-37 in serum.Methods: Mouse anti-human IL-37 monoclonal antibody was used as capturing antibody,rabbit anti-human IL-37 polyclonal antibodies served as detection antibody,HRP labeled goat anti-rabbit IgG employed as second antibody and recombinant human IL-37 protein used as reference standard for the establishment of a doubleantibody sandwich ELISA.The working conditions were optimized,such as sensitivity,linear range,reproducibility and evaluated the serum IL-37 in patients with Dengue fever.Results: The sensitivity of the established ELISA method was 1.465 μg/L approximately.Likewise,the linearity range of this method was about (1.465-46.875) μg/L.Further,the co efficient of variation (CV) of inter-batch and intra-batch in this study were 6.6% and 11.7%,respectively.Notably,this method could be used in the detection of IL-37 in serum of the patients with Dengue fever,showing that the level of IL-37 in Dengue fever patients was much higher than that in healthy controls.Conclusion: The double antibody sandwich ELISA assay for the detection of human IL-37 was successfully established,which can be apply to detect of human IL-37 in clinical samples.