目的研制特异性检测人可溶性OX40L蛋白(sOX40L)的ELISA试剂盒,探讨其在自身免疫性疾病临床诊断中的应用。方法利用两株识别不同抗原表位的鼠抗人OX40L单克隆抗体1G1和4C12分别作为包被抗体和检测抗体,采用双抗体酶联免疫夹心法研制特异性检测人sOX40L的ELISA方法,并对其稳定性、精确性和特异性进行分析。结果①建立的检测人sOX40L酶联免疫试剂盒在抗原浓度为3.91~250ng/ml范围内有良好的线性关系,并具有良好的稳定性、较高的准确性和特异性。②利用该ELISA方法检测到sOX40L蛋白在肾病综合征和Graves’病患者血清中表达水平均显著高于健康对照组。结论成功建立了检测人sOX40L蛋白的ELISA试剂盒,应用该方法检测到sOX40L在多种自身免疫病患者血清中异常高表达。该试剂盒在OX40L分子基础研究以及自身免疫病的临床诊断方面具有潜在的应用价值。
Objective To establish a ELISA assay for evaluating soluble human OX40L protein and evaluate its clinical values in autoimmune diseases.Methods The sandwich ELISA was prepared by utilizing two anti -hOX40L monoclonal antibodies 1G1 and 4C12 as capture and detecting antibody respectively.Then its stability,precision and specificity were analyzed.Results A ELISA assay for the detection of soluble OX40L with high stability,veracity and specificity was developed.The detection range of the ELISA permits a reliable assay of sOX40L from 3.91~250ng/ml.Compared with healthy control group,high expression levels were detectable in the sera of patient group of Graves' disease and nephritic syndrome.Conclusion By using the ELISA kit,high sOX40L level in the sera of patient with autoimmune diseases was detected.It may be of significance in basic research of OX40L and diagnosis and prognosis estimation of autoimmune diseases.