重组子宫内膜抗原制备及其在蛋白芯片检测中的应用

Preparation of recombinant endometrial antigen and its application in protein chip detection

目的探索一种纯度高、简便、低成本制备抗子宫内膜抗原的系统和便捷的检测方法。方法通过YK537大肠杆菌表达系统制备人重组子宫内膜抗原蛋白。获得蛋白制备蛋白芯片,利用蛋白芯片方法和酶联免疫吸附法(ELISA)方法检测71例不孕患者抗子宫内膜抗体。结果重组子宫内膜蛋白基因序列克隆入表达载体phoA下游并在大肠杆菌中成功表达。SDS-PAGE和HPLC的结果显示重组子宫内膜抗原蛋白纯度大于95%。通过蛋白芯片和ELISA检测显示蛋白芯片和传统ELISA方法的受试者工作特征曲线(ROC曲线)显示结果一致,曲线下面积为0.89。结论新设计的重组子宫内膜抗原蛋白能在大肠杆菌分泌表达系统简便、高效地表达,将获得的新抗原用于蛋白芯片,操作简便,结果可信,有望成为临床检测抗子宫内膜抗体的手段。

Objective：To prepare a high purity,simple,low cost anti-endometrial antigen system and a convenient detection method Methods： Recombinant human endometrium antigen was prepared by YK537 Escherichia coli expression system.The protein chip of the protein prepared was prepared and the anti-endometrium antibody of 71 infertile patients was detected by ELISA method.Results：The recombinant endometrial protein genes sequences were cloned into the downstream of expression vector phoA,and the proteins were successfully expressed in Escherichia coli.The results of SDS-PAGE and HPLC showed that the purity of recombinant endometrial proteins was more than 95%.The results of protein chip and ELISA showed that the receiver operating characteristic curve（ROC）of the protein chip was consistent with the traditional ELISA,and the area under the curve was 0.89.Conclusions：The newly designed recombination endometrium antigen can be expressed easily and efficiently in the E.coli secretory expression system.The protein chip detection method is easy to operate,and it is expected to become a means of clinical detection of anti-endometrial antibodies.