全人源抗狂犬病病毒scFv-Fc融合抗体的制备及活性分析
作者:
作者单位:

作者简介:

通讯作者:

中图分类号:

基金项目:

国家自然科学基金资助(81273325);江苏省科技支撑计划资助项目(BE2011842)


Preparation and activity analysis of a whole human scFv-Fc fusion antibody against rabies virus
Author:
Affiliation:

Fund Project:

  • 摘要
  • |
  • 图/表
  • |
  • 访问统计
  • |
  • 参考文献
  • |
  • 相似文献
  • |
  • 引证文献
  • |
  • 资源附件
  • |
  • 文章评论
    摘要:

    目的:制备全人源抗狂犬病病毒scFv-Fc融合抗体,并分析其结合活性-方法:构建全人源抗狂犬病病毒scFv-Fc融合抗体原核表达载体,优化表达并纯化全人源抗狂犬病病毒scFv-Fc融合抗体-以纯化的灭活狂犬病病毒包板,对纯化的抗体进行结合活性分析-结果:构建全人源抗狂犬病病毒scFv-Fc-pColdⅡ原核表达载体,经测序分析正确后,转入大肠杆菌BL21(DE3)进行表达,在加入浓度为0.5 mmol/L IPTG时,scFv-Fc融合抗体的表达较高,其分子量大小为57 000-纯化后的scFv-Fc融合抗体在1∶512的条件下仍可以较好地结合狂犬病病毒-结论:构建全人源抗狂犬病病毒scFv-Fc-pColdⅡ原核表达载体,表达并纯化了scFv-Fc融合抗体,为研发狂犬病暴露后预防治疗性抗体药物奠定了基础-

    Abstract:

    Objective:To prepare a human scFv-Fc fusion antibody against rabies virus and to analyze its binding activity. Methods:The human scFv-Fc prokaryotic expression vector was constructed. After optimizing prokaryotic expression system,the human scFv-Fc fusion antibody against rabies virus was expressed and purified. Purified humanized scFv-Fc antibody was confirmed its binding activity by binding to purified inactivated rabies virus. Results:After sequence analysis,the human scFv-Fc-pColdⅡprokaryotic expression vector was successfully established. Through transforming E.coli.BL21 (DE3),human scFv-Fc fusion antibody was expressed by optimized induction of IPTG at concentration of 0.5 mmol/L. The expression of scFv-Fc fusion antibody was increased and its molecular weight was 57 000. By purification,the human scFv-Fc fusion antibody was purified with excellent binding activity even in 512 times diluted concentration. Conclusion:We have successfully established a whole human anti-rabies virus scFv-Fc-pColdⅡ prokaryotic expression vector,and expressed and purified scFv-Fc fusion antibody. This fusion antibody can lay a foundation for developing novel antibody-targeted drugs of rabies prophylaxis after exposure.

    参考文献
    相似文献
    引证文献
引用本文

高 畅,张倩倩,熊四平,唐小军,仇镇宁,冯振卿,朱 进.全人源抗狂犬病病毒scFv-Fc融合抗体的制备及活性分析[J].南京医科大学学报(自然科学版),2014,(6):741-744-749

复制
分享
文章指标
  • 点击次数:
  • 下载次数:
  • HTML阅读次数:
  • 引用次数:
历史
  • 收稿日期:2014-01-16
  • 最后修改日期:
  • 录用日期:
  • 在线发布日期: 2014-06-19
  • 出版日期:
通知关闭
郑重声明