Gao Pu-Jun, Piao Yun-Feng, Liu Xiao-Dong, Qu Li-Ke, Shi Yang, Wang Xiao-Cong, Yang Han-Yi
Department of Digestion, 1st Hospital affiliated to Jilin University, Changchun 130021, Jilin Province, China.
World J Gastroenterol. 2003 Sep;9(9):2114-6. doi: 10.3748/wjg.v9.i9.2114.
To observe the binding activity of beta-2-glycoprotein I(beta(2)GPI) to hepatitis B surface antigen (HBsAg) and the possible roles of beta(2)GPI in hepatitis B virus (HBV) infection.
The rationale of ELISA methods and ELISA-based research method and ligand-blotting technique were used to detect the specific interaction of beta(2)GPI with HBsAg.
With the increase of rHBsAg, the binding of beta(2)GPI to rHBsAg elevated, and these changes had statistic significance. When we added non- biotinlyated beta(2)GPI, the OD value significantly decreased though they still were positively relevant to rHBsAg, suggesting non- biotinlyated beta(2)GPI competed with biotinlyated beta(2)GPI to saturate the binding sites on rHBsAg. Meanwhile BSA was used as negative control to substitute for rHBsAg coating the plates. The results indicated no interaction between beta(2)GPI and BSA, suggesting the affinity of beta(2)GPI to rHBsAg was specific. The ligand blotling indicated that beta(2)GPI might bind to rHBsAg no matter whether it was under reduced condition or not.
The binding of beta(2)GPI to HBsAg suggests that beta(2)GPI may be a carrier of HBV and that beta(2)GPI may play important roles in HBV infection.
观察β2糖蛋白I(β2GPI)与乙型肝炎表面抗原(HBsAg)的结合活性以及β2GPI在乙型肝炎病毒(HBV)感染中的可能作用。
采用酶联免疫吸附测定(ELISA)方法的原理、基于ELISA的研究方法和配体印迹技术检测β2GPI与HBsAg的特异性相互作用。
随着重组乙型肝炎表面抗原(rHBsAg)浓度增加,β2GPI与rHBsAg的结合增加,且这些变化具有统计学意义。当加入未生物素化的β2GPI时,光密度(OD)值显著降低,尽管其仍与rHBsAg呈正相关,提示未生物素化的β2GPI与生物素化的β2GPI竞争以饱和rHBsAg上的结合位点。同时,用牛血清白蛋白(BSA)作为阴性对照替代rHBsAg包被酶标板。结果表明β2GPI与BSA之间无相互作用,提示β2GPI与rHBsAg的亲和力具有特异性。配体印迹显示,无论是否处于还原条件下,β2GPI均可能与rHBsAg结合。
β2GPI与HBsAg的结合提示β2GPI可能是HBV的载体,且β2GPI可能在HBV感染中发挥重要作用。