卡波济肉瘤相关疱疹病毒包膜糖蛋白K8.1单克隆抗体的鉴定

目的:对已制备并经初筛的卡波济肉瘤相关疱疹病毒(KSHV)包膜糖蛋白K8.1单克隆抗体进行特异性鉴定。方法:采用酶联免疫吸附试验(ELISA),经3次克隆选择,筛选出能稳定分泌针对K8.1蛋白单克隆抗体(McAb)的杂交瘤细胞株共9株。采用Westernblot法评价9株杂交瘤细胞培养上清识别原核表达重组K8.1蛋白和原发性渗出性淋巴瘤(PEL)细胞系BCBL-1中KSHVK8.1包膜糖蛋白的能力。结果:筛选了9株能够稳定分泌抗K8.1单克隆抗体的杂交瘤细胞株;Westernblot法显示,9株单抗均能特异性识别原核表达K8.1/GST融合蛋白和BCBL-1中KSHVK8.1包膜糖蛋白。结论:成功制备了KSHV包膜糖蛋白K8.1单克隆抗体。

Characterization of monoclonal antibody against envelope glycoprotein K8.1 gene of kaposi's sarcoma-associated herpesvirus (KSHV)

Objective: To identify the specification of the monoclonal antibody(McAb) against the envelope glycoprotein K8.1 of Kaposi's sarcoma-associated herpesvirus(KSHV).Methods: By way of three times of cloning choice,enzyme linked immunosorbent assay(ELISA) was performed to generate nine cell lines which stably produced McAbs against K8.1 protein.Furthermore,the specificity of McAbs was identified by Western blot.Results: Nine cell lines which stably produced McAbs against K8.1 protein were screened successfully by using ELISA assay.Western blot analysis demonstrated that McAbs produced by nine cell lines could specifically recognize the prokaryotic expressed fusion protein K8.1/GST and the envelope glycoprotein K8.1 of KSHV in BCBL-1.Conclusion: McAbs against the envelope glycoprotein K8.1 of KSHV were prepared successfully.