脊髓灰质炎病毒Ⅰ型单克隆抗体的制备  

Preparation of Monoclonal Antibody for Type Ⅰ Poliovirus

作　　者：王朝元[1] 唐永洪 黄圣 尹玉莹 杨继红[2] Wang Chaoyuan;Tang Yonghong;Huang Sheng;Ying Yuying;Yang Jihong(College of Life Sciences,South-Central University for Nationalities,Wuhan 430074,China;College of Life Sciences,Huazhong Normal University,Wuhan 430079,China)

机构地区：[1]中南民族大学生命科学学院,武汉430074 [2]华中师范大学生命科学学院,武汉430079

出　　处：《中南民族大学学报（自然科学版）》2018年第4期35-39,共5页Journal of South-Central University for Nationalities：Natural Science Edition

基　　金：中央高校基本科研业务费专项资金资助项目(37001/CZP17010);中南民族大学横向科研资助项目(HZY17059)

摘　　要：目的:为建立脊灰病毒Sabin株Ⅰ型特异的单克隆抗体.方法:以Vero细胞增殖病毒并用蔗糖密度梯度离心法纯化后,用病毒免疫SPF级Balb/c小鼠.取小鼠免疫脾淋巴细胞与SP2/0-Ag14骨髓瘤细胞进行细胞融合,通过间接ELISA法筛选抗体阳性的杂交瘤细胞,并用有限稀释法克隆化.用细胞培养法制备脊灰病毒单抗,并以中和试验测定单抗效价.结果:获得了8株分泌抗脊灰病毒Sabin株Ⅰ型单克隆抗体的杂交瘤细胞株,其中1株G8G7能够稳定分泌抗体.间接ELISA法测得单抗的效价为1:8,中和试验测得中和抗体的效价为1:58.结论:G8G7分泌的抗体为抗脊灰病毒Sabin株Ⅰ型特异的单克隆抗体.Objective: To establish a specific monoclonal antibody of poliovirus Sabin strain Ⅰ. Methods: After Poliovirus Sabin strain type Ⅰ was proliferated in Vero cells and purified by sucrose density gradient centrifugation,SPF Balb/c mice were immunized by poliovirus. Then the splenic lymphocytes from immunized mice were fused with SP2/0-Ag14 myeloma cells. The positive hybridoma cells were screened by indirect ELISA and cloned by limited dilution method. The monoclonal antibody of poliovirus type Ⅰ was prepared by cell culture and its titer was determined by neutralization test. Results: Eight stable hybridoma cell lines secreting anti-poliovirus Sabin strain I monoclonal antibodies were obtained,one of which( G8G7) could secret antibodies stably. The titer of monoclonal antibody measured by indirect ELISA was 1:8,and the titer of neutralized antibody measured by neutralization test was 1 : 58. Conclusion: It is concluded that the antibody secreted by G8G7 hybridoma cell lines is a type Ⅰ-specific monoclonal antibody against poliovirus Sabin strain.

关 键 词：单克隆抗体 脊髓灰质病毒Ⅰ型 Vero细胞系 

分 类 号：Q939.91[生物学—微生物学]