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应用一种人巨细胞病毒(CMV)抗体CMVB1的抗独特型单克隆抗体(McAb8C9)建立了一种人类巨细胞病毒(HCMV)新的ELISA检测法。CMVB1是一种HCMV糖蛋白抗原复合物(gB)的特异性抗体,用交联于载体上的CMVB1免疫纯系BALB/c小鼠后,取其脾细胞制备杂交瘤细胞,经筛选获得了抗独特型抗体McAb-8C9,再经纯化后应用。试验中所用CMVB1用生物素-XNHS标记,生物素/蛋白分子比率为10:1,所用HCMV为原型株AD169(2.2×10~5pfu/ml,100μlHCMV+1ml正常尿),用前95℃加热10min灭活。用每孔0.5μl的抗独特型抗体室温过夜包被微量板,PBS-Tween20洗涤后用1%BSA/PBS封闭,加入37℃预孵育
A novel ELISA assay for human cytomegalovirus (HCMV) was developed using an anti-idiotypic monoclonal antibody (McAb 8C9), a human cytomegalovirus (CMV) antibody. CMVB1 is a specific antibody against HCMV glycoprotein antigen complex (gB). BALB / c mice were immunized with CMVB1 cross-linked to the vector and splenocytes were harvested to prepare hybridoma cells. Anti-CMVB1 Unique antibody McAb-8C9, and then applied after purification. The CMVB1 used in the experiment was labeled with biotin-XNHS and the biotin / protein ratio was 10: 1. The HCMV used was the original strain AD169 (2.2 × 10 ~ 5pfu / ml, 100μl HCMV + 1ml normal urine) Inactivated Microtiter plates were coated with 0.5 μl anti-idiotypic antibody per well overnight at room temperature, washed with PBS-Tween 20, blocked with 1% BSA / PBS and pre-incubated at 37 ° C