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
投稿时间:2018-05-16  修订日期:2018-05-16
DOI:
中文关键词: 单克隆抗体  脊髓灰质病毒I型
英文关键词: monoclonal antibody  type I poliovirus
基金项目:脊髓灰质炎病毒I型单克隆抗体制备(37001/CZP17010)
作者单位E-mail
王朝元 中南民族大学 生命科学学院 wangchaoyuan@mail.scuec.edu.cn 
唐永洪 中南民族大学 生命科学学院  
黄圣 中南民族大学 生命科学学院  
杨继红 华中师范大学 生命科学学院 705772657@qq.com 
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中文摘要:
      建立脊灰病毒Sabin株I型特异的单克隆抗体。利用Vero细胞增殖脊灰病毒Sabin株I型,用蔗糖密度梯度离心法纯化的病毒免疫SPF级Balb/c小鼠,取其免疫脾淋巴细胞与SP2/0-Ag14骨髓瘤细胞进行细胞融合,通过建立的间接ELISA筛选抗体阳性的杂交瘤细胞。阳性杂交瘤细胞通过有限稀释法克隆化,细胞培养法制备单抗,中和试验测定抗体效价。获得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;细胞培养法制备的单抗,利用间接ELISA测得抗体效价为1:8,中和试验测得中和抗体效价为1:58。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:
      The monoclonal antibody for type I poliovirus were prepared. Vero cells were used to proliferate poliovirus. Poliovirus Sabin strain type I, which was purified by sucrose density gradient centrifugation, was used to immunize Balb/c mice, and the spleen cells of the immunized mice were fused with mouse SP2/0-Ag14 myeloma cells. The hybridoma cells stably secreting monoclonal antibodies type I poliovirus were screened by indirect ELISA and then cloned by limited dilution method. One stable hybridoma cell line G8G7 were obtained. The titer of the monoclonal antibodies secreted by cell culture, measured with indirect ELISA assay was 1/8, and neutralization antibody titer measured by neutralization test was 1/58. We have 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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