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目的制备淋巴细胞脉络丛脑膜炎病毒(LCMV)单克隆抗体(m Ab)。方法采用杂交技术获得并筛选出杂交瘤细胞,收集含单克隆抗体的培养上清行免疫荧光(IFA)、dot-blot、ELISA法鉴定抗体重链类型和测定单克隆抗体的免疫效价。用蛋白G Sepharose亲和层析填料纯化抗体,纯化后的抗体采用SDS-PAGE检测纯化效果,并进一步用Western blot检查抗体对LCMV的作用部位。结果成功获得能稳定分泌抗LCMV的特异性杂交瘤细胞株,顺利获得纯化的单克隆抗体,命名为anti-GP2。免疫荧光、ELISA结果均证实获得的单克隆抗体能识别LCMV抗原。用ELISA法鉴定出该单克隆抗体重链类型为Ig G2b,培养液免疫效价大约为1.35×10~3。蛋白G Sepharose亲和层析填料纯化抗体后经SDS-PAGE检测证实纯化抗体成功。Western blot提示该单克隆抗体对LCMV的作用部位为病毒颗粒表面糖蛋白抗原GP2。结论成功制备出了针对LCMV病毒颗粒表面糖蛋白抗原GP2的单克隆抗体(anti-GP2 m Ab)。
Objective To prepare a monoclonal antibody against lymphocytic choriomeningitis (LCMV) (m Ab). Methods The hybridomas were obtained and screened by hybridization. Immunofluorescence (IFA), dot-blot and ELISA were used to identify the type of antibody heavy chain and the immunological titer of the monoclonal antibody. Antibodies were purified with Protein G Sepharose affinity chromatography. Purified antibodies were tested for their purification by SDS-PAGE and Western blotting for antibodies against LCMV. Results The specific hybridoma cell line that can stably secrete anti-LCMV was successfully obtained. The purified monoclonal antibody was successfully obtained and named anti-GP2. Immunofluorescence and ELISA results confirmed that the obtained monoclonal antibody recognized LCMV antigen. The heavy chain of this monoclonal antibody was identified as Ig G2b by ELISA. The immune titer of the monoclonal antibody was about 1.35 × 10 ~ 3. Protein G Sepharose affinity chromatography packing purified antibody SDS-PAGE test confirmed the successful purification of antibodies. Western blot suggested that the site of action of this monoclonal antibody on LCMV was GP2, a glycoprotein antigen. Conclusion Anti-GP2 m Ab was successfully prepared against the glycoprotein antigen GP2 of LCMV.