肾综合征出血热病毒84-Fli株单克隆抗体的制备及鉴定

Preparation and identification of monoclonal antibody against hemorrhagic fever with renal syndrome virus 84-Fli strain

目的制备抗肾综合征出血热(hemorrhagic fever with renal syndromes,HFRS)病毒特异性单克隆抗体。方法以汉坦病毒84-Fli株抗原纯化液免疫BALB/c小鼠,采用细胞融合技术获得单克隆抗体,利用IFA、SDS-PAGE和Western blot法鉴定其生物学特性。结果成功筛选出阳性杂交瘤细胞株4F6,能稳定且持续分泌抗HFRS病毒的单克隆抗体,亚类鉴定结果为IgM,可与HFRS病毒蛋白在相对分子质量约50 000处发生特异性反应,具有较好的专属性。结论成功制备了1株抗HFRS病毒的单克隆抗体,为进一步研究该病毒感染的诊断、治疗及其预防方法奠定了基础。

Objective To prepare the specific monoclonal antibody against hemorrhagic fever with renal syndrome（HFRS）virus. Methods BALB/c mice were immunized with purified liquid of Hantaan virus 84-Fli strain, based on which monoclonal antibodies were prepared by cell fusion technology, and identified for biological characters by IFA, SDS-PAGE and Western blot. Results A positive hybridoma cell strain 4F6 was successfully screened, which stably and persistently secreted monoclonal antibody against HFRS virus, was identified as subtype IgM, and showed specific reaction band, with a relative molecular mass of about 50 000, with HFRS virus protein, indicating a high specificity. Conclusion A monoclonal antibody against HFRS virus was successfully prepared, which laid a foundation of further study on diagnosis, therapy and prevention of infection with HFRS virus.