人5型腺病毒六邻体蛋白单克隆抗体的制备及鉴定

Preparation and identification of specific monoclonal antibodies against hexon of HAdV5

目的制备抗人5型腺病毒(HAdV5)六邻体(Hexon)蛋白的单克隆抗体并对其进行鉴定。方法用纯化的Hexon蛋白免疫BALB/c小鼠,取其脾细胞与骨髓瘤细胞融合,经ELISA间接法筛选和克隆化培养,获得4株分泌抗HAdV5的Hexon蛋白单克隆抗体的杂交瘤细胞。纯化获得单克隆抗体,使用ELISA和Western blot鉴定单克隆抗体的敏感性和特异性。结果 ELISA和Western blot结果表明这4株单克隆抗体与HAdV5的Hexon蛋白可以特异性结合。单克隆抗体株4A9-HRP标记和8D6建立的ELISA夹心法具备较高灵敏度和特异性。结论获得4株抗HAdV5的Hexon蛋白的单克隆抗体,为HAdV5相关的疫苗研制和HAdV5感染性疾病的早期快速诊断奠定了基础。

Objective To obtain the monoclonal antibody against hexon protein of human adenovirus type 5 （HAdVS）. Methods BALB/c mice were immunized with purified hexon protein, and the spleen cells of the mice were isolated and fused with myloma cells. Four hybridoma cell strains were screened by indirect ELISA and cul- tured,and the sensitivity and specificity were analyzed by ELISA and Western blotting test. Results These hybrido- ma cells contained specific monoclonal antibodies against hexon protein of HAdV5 as identified by ELISA and West- ern blot. Conclusion The monoclonal antibodies against hexon protein of HAdV5 with high specificity have been successfully obtained,and these antibodies could be useful in developing assays for early diagnosis of HAdV5 infec- tion and also in vaccine development based on HAdV5 virus.