检测人IL-37双抗夹心ELISA方法的建立

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

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Ig G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1.465μg/L,线性范围为1.465~46.875μg/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。

ObjectiveTo establish a double antibody sandw-ich 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 sandw-ich ELISA. The w-orking conditions were optimized,such as sensitivity,linear range,reproducibility and evaluated the serum IL-37 in patients wdth Dengue fever. Results： The sensitivity of the established ELISA method was 1.465 滋 g^^L approximately. Likew-ise,the linearity range of this method was about （1.465-46.875）滋 g/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 w dth Dengue fever, show-ing that the level of IL-37 in Dengue fever patients was much higher than that in healthy controls. Conclusion： The double antibody sandw- ich ELISA assay for the detection of human IL-37 was successfully established, w-hich can be apply to detect of human IL-37 in clinical samples.