人源全分子抗c-Met 抗体的制备及对鼻咽癌细胞生物学特性的影响
作者:
作者单位:

作者简介:

通讯作者:

中图分类号:

基金项目:

江苏省科技厅临床医学科技专项计划(BL2013038);国家自然科学基金(81502673)


Preparation of full molecule human anti-c-Met antibody and its effects on nasopharyngeal carcinoma cells
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:利用基因工程技术制备全分子人源抗 c-Met抗体,分析该抗体免疫学特性,并观察该全分子抗体对鼻咽癌细胞增殖-迁移和侵袭的影响-方法:设计引物扩增抗c-Met Fab抗体的重轻链可变区序列,将其分别克隆到真核表达载体pFUSE-CHIg-hG1 和 pFUSE-CLIg-hκ 中-转染真核细胞,表达产物用Protein A柱纯化-通过ELISA-Western blot和免疫荧光等方法鉴定该抗体的免疫学特性-CCK-8检测抗体对人鼻咽癌细胞增殖的抑制作用,划痕实验-Transwell侵袭实验分析抗体对人鼻咽癌细胞迁移和侵袭的影响-结果:成功重组全分子人源抗c-Met 抗体的表达载体,获得的全分子抗体保留了与抗原的特异性结合活性-CCK-8实验中结果显示全分子抗体能抑制c-Met阳性鼻咽癌细胞CNE2和HONE1的增殖-细胞划痕和Transwell侵袭实验结果显示全分子抗体能抑制c-Met阳性细胞CNE2和HONE1的迁移和侵袭能力-结论:本研究成功制备人源全分子抗c-Met抗体,该抗体有明显的中和活性,为探索鼻咽癌分子靶向治疗奠定了基础-

    Abstract:

    Objective:To produce a full molecule human anti-c-Met antibody with genetic engineering, and analyze the biological activity of the antibody and observe its influence on proliferation, migration and invasion of nasopharyngeal carcinoma cells. Methods:We designed a primer for amplifying the variable region of anti-c-Met Fab antibody. VH and VL genes were cloned into pFUSE-CHIg-hG1 and pFUSE-CLIg-hκ expression vectors, respectively, and were both transfected into eukaryotic cells. The expression product was purified using protein A column. The identification and immunological characteristics of antibody were analyzed by enzyme-linked immune sorbent assay (ELISA), Western blotting assay and immunofluorescence assay. The inhibition function of the antibody in nasopharyngeal carcinoma cells was analyzed by CCK-8. The effects of the antibody on the invasion and migration of nasopharyngeal carcinoma cells was determined by wound-healing assay and Transwell invasion assay. Results:The results demonstrated that the full molecule human anti-c-Met antibody was successfully produced, and the antibody effectively recognized c-Met protein. In nasopharyngeal carcinoma cell experiment, CCK8 assay demonstrated that the antibody played a significant role in proliferation of CNE2 and HONE1. Wound-healing assay and Transwell invasion assay demonstrated that the antibody could suppressed the migration and invasion of CNE2 and HONE1. Conclusion:In this study, full molecule human anti-c-Met antibody was established successfully and has obvious neutralizing effect. The antibody is expected to become a candidate molecule for molecular targeted therapy of nasopharyngeal carcinoma.

    参考文献
    相似文献
    引证文献
引用本文

白璐月,刘金荣,唐 奇,熊四平,陈 雅,杜银娟,周 荧,朱 进,陈仁杰.人源全分子抗c-Met 抗体的制备及对鼻咽癌细胞生物学特性的影响[J].南京医科大学学报(自然科学版),2016,(6):687-692

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:2015-11-03
  • 最后修改日期:
  • 录用日期:
  • 在线发布日期: 2016-06-15
  • 出版日期: