生物学杂志

生物学杂志

• 技术方法 • 上一篇    下一篇

抗人TDP-43单克隆抗体的制备纯化及效价测定

  

  1. 1. 丽水市第二人民医院, 丽水 323000; 2. 杭州浙大迪迅生物基因工程有限公司， 杭州 310052
  • 出版日期:2018-08-18 发布日期:2018-08-18
  • 通讯作者: 刘奕，硕士研究生，从事免疫学研究方向，E-mail：13556754@qq.com
  • 作者简介:吴绍长，硕士研究生，主任医师，从事神经内科研究方向，E-mail：1473383870@qq.com
  • 基金资助:
    丽水市科技计划项目（2013JYZB61）

Preparation and purification of monoclonal antibody against human TDP-43#br#

  1. 1. Lishui Second People′s Hospital， Lishui 323000；2. Hangzhou Zheda Dixun Biological Gene Engineering Co., Ltd, Hangzhou 310052, China
  • Online:2018-08-18 Published:2018-08-18

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摘要： 为了建立人TDP-43蛋白的检测、纯化方法以及为开发相关的检测试剂提供基础，利用重组TDP-43免疫BALB/c小鼠，采用杂交瘤细胞技术，获得分泌抗人TDP-43蛋白抗体的杂交瘤细胞株，用杂交瘤细胞株诱生小鼠腹水，应用蛋白A亲和层析法进行抗体纯化，并采用ELISA测定抗体效价。结果表明，2株稳定分泌抗人TDP-43单克隆抗体细胞株1C5和2A5被成功制备，其亚类均为IgG1,效价分别为1∶ 256 000和1∶ 512 000。

关键词:  , TDP-43；单克隆抗体；制备；鉴定

Abstract: In order to establish a detection method and purification technology of human TDP-43 for the development of relevant test kits, BALB/c mice were immunized with purified recombinant human TDP-43, hybridoma cell lines secreting monoclonal antibodies (McAbs) against human TDP-43 were obtained by hybridoma technique. The McAbs were purified using affinity chromatography on immobilized protein A and identified by their subtype and titers via ELISA. The results showed that two monoclonal antibodies of 1C5 and 2A5 belonging to IgG1 were successfully prepared with the titer of 1∶ 256 000 and 1∶ 512 000.

Key words: TDP-43, monoclonal antibody, preparation, characterization