王朝元,唐永洪,黄 圣,尹玉莹,杨继红.脊髓灰质炎病毒I型单克隆抗体的制备[J].中南民族大学学报自然科学版,2018,(4):35-39
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
  
DOI:10.12130/znmdzk.20180408
中文关键词: 单克隆抗体  脊髓灰质病毒 I 型  Vero 细胞系
英文关键词: monoclonal antibody  type I poliovirus  Vero cell lines
基金项目: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)
作者单位
王朝元1,唐永洪1,黄 圣1,尹玉莹1,杨继红2 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 
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中文摘要:
      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.
英文摘要:
      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.
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