具有中和活性抗人类腺病毒单克隆抗体的制备、鉴定及应用  

Preparation,characterization and application of anti-human adenovirus monoclonal antibody with neutralization activity

作　　者：李思袖[1] 张国成[1] 许东亮[1] 聂晓晶[1] 李小青[1] 汪志华[1] 张学红[1] 

机构地区：[1]第四军医大学西京医院儿科,陕西西安710032

出　　处：《细胞与分子免疫学杂志》2009年第12期1140-1142,共3页Chinese Journal of Cellular and Molecular Immunology

摘　　要：目的:制备具有中和活性抗人类腺病毒(HAdv)单克隆抗体(mAb),并进行鉴定与应用。方法:以活的人类腺病毒3型(HAdv-3)滴鼻免疫BALB/c鼠,用细胞融合技术制备抗HAdv的mAb细胞株,采用中期分裂相秋水仙素阻抑法对mAb细胞株染色体进行分析;使用鼠mAb亚型鉴定试剂盒进行抗体亚型鉴定,通过ELISA、Western blot和间接免疫荧光技术进行特异性鉴定。建立HAdv-3感染动物模型,用获得的HAdv-3 mAb进行保护性研究。结果:细胞融合率为86%,抗HAdv抗体阳性孔率为51.4%。鉴定1株杂交瘤细胞(1A4),染色体数为98条,抗体亚类属IgG2a/κ,抗体腹水ELISA效价达10-5。ELISA、Western blot和间接免疫荧光证实该mAb特异性好。该mAb对HAdv-3感染动物有保护性作用。结论:成功地制备具有中和活性的抗HAdv mAb。该mAb识别的是HAdv六邻体蛋白,对HAdv-3感染动物有保护作用。AIM： To prepare, identify and apply antihuman adenoviru（HAdv） neutralization monoclonal antibody （mAb）. METHODS： BALB/c mice were immunized with live human adenovirus type3 （HAdv-3） strain intranarially. Sp2/0 cells were fused with the spleen cells harvested from BALB/c mice. The chromosomal amounts of the hybridoma cells were analyzed by colchicine. A commercially available mouse mAb isotyping kit was used to identify the isotype of this mAb. Clones secreting specific monoclonal antibody were screened by indirect enzyme linked immunosorbent assay （ELISA）, Western blot and indirect immunofluorescent assay. Infected animal model was established, and the protective effect of mAb was studied. RESULTS： The fusion rate was 86%, and the positive rate was 51.4%. One of the hybridoma cell was identified （IA4）, the chromosomal amounts of was 98, the subtype mAb type belonged to IgG2a/K, and the titer of the mAb secreted by the strain in ascite reached more than 10-5. The specificity of the mAb was proved by ELISA , Western blot and indirect immunofluorescent assay. This mAb had protective effect on animal infected by HAdv-3. CONCLUSION： The anti-adenovirus mAb which have neutralization activities was successfully prepared. The mAb recognized the hexon subunit and had protective effect on animal infected by HAdv-3.

关 键 词：腺病毒 单克隆抗体 中和活性 

分 类 号：Q813.2[生物学—生物工程]