抗人类疱疹病毒7型南京分离株YY5单克隆抗体的制备与鉴定

Preparation and Characterization of Monoclonal Antibodies Against Human Herpesvirus 7

目的:制备抗人类疱疹病毒7型(HHV-7)单克隆抗体,应用于临床与科研。方法:用纯化的HHV-7南京株YY5免疫Balb/c小鼠,取其脾细胞与骨髓瘤细胞SP2/0融合,经间接ELISA法筛选。间接免疫荧光、免疫印迹试验等方法鉴定单抗。结果:获得了3株分泌抗(HHV-7)单克隆抗体的杂交瘤细胞系,分别命名为6H9、6C11、4D5。单克隆抗体亚类鉴定表明:6H9为IgG1类,6C11、4D5为IgM类。用间接ELISA方法检测单抗腹水滴度分别为:1:6.40×104,1:1.28×105,1:1.00×103。免疫印迹试验结果表明:6C11能与分子质量约70 ku大小的病毒蛋白结合。结论:成功制备并鉴定3株抗HHV-7单克隆抗体,为进一步研究HHV-7的特性及为临床快速诊断提供可能。

Objective: To prepare monoclonal antibodies against human herpesvirus 7 (HHV-7). Methods: Purified HHV-7 strain YY5 was immunized to Balb/c mouse, and the spleen cells of the immunized mouse were fused with SP2/0 myeloma cells. The positive clones which produced McAbs against HHV-7 were screened by ELISA. Results: Clones named as 6H9, 6C11, 4D5 secreting specific McAbs against HHV-7 were obtained. Among these, 6H9 was of the IgG1. 6C11 and 4D5 were of the IgM, Ascitic titres of the antibodies were 1: 6. 40× 104, 1: 1. 28× 105, 1: 1. 00 ×103, when tested by ELBA respectively. Western-blot showed that 6C11 was bound to 70ku protein of HHV-7. Conclusions: The McAbs against HHV-7 may be useful tool for studying biological properties of HHV-7 and clinic laboratory detection.