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Mammalian cell display for rapid screening scFv antibody therapy
  • ISSN号:1672-9145
  • 期刊名称:《生物化学与生物物理学报:英文版》
  • 时间:0
  • 分类:Q78[生物学—分子生物学] TN873[电子电信—信息与通信工程]
  • 作者机构:[1]Beijing Institute of Radiation Medicine, Beijing 100850, China, [2]Anhui Medical University, Hefei 230032, China, [3]Beijing Institute of Microbiology and Epidemiology, Beijing 100071, China, [4]Chinese PLA General Hospital, Tumor Center Laboratory, Beijing 100850, China
  • 相关基金:This work was supported by the grants from the Beijing Natural Science Foundation (5113034) and the National Natural Science Foundation of China (81272701).
中文摘要:

人的抗体开始在有免疫力的基因治疗为使用正在引起注意。对癌症和传染疾病的治疗合适的有效治疗学的 monoclonal 抗体的有效产生将是极其珍贵的。抗体显示方法逐渐地被用来屏蔽人的 monoclonal 抗体。这里,我们报导一个哺乳动物的房间显示方法的构造源于一个天真的抗体全部剧目,为人,单个链的变量碎裂短暂地基于 pDisplay 向量在 293T 房间表面上被显示了。当前的 pDisplayscFv 抗体全部剧目的尺寸被估计了是 0.74

英文摘要:

Human antibodies are beginning to draw attention for use in immune gene therapy. The efficient generation of effective therapeutic monoclonal antibodies suitable for the treatment of cancers and infectious diseases would be enormously valuable. Antibody display methods are increasingly used to screen human monoclonal antibodies. Here we report the construction of a mammalian cell display method derived from a naive antibody repertoire, for which human single- chain variable fragments (scFv) have been transiently dis- played on 293T cell surfaces based on a pDisplay vector. The sizes of the current pDisplay-scFv antibody repertoires have been estimated to be 0.74 × 10^7. An immunoblot assay confirmed the expression of the scFv antibody library. The subceilular distribution of ErbB3-scFv expression plasmid facilitated the display of ErbB3 scFv on the cell membrane surface and the efficiency of the display was evaluated by fluorescence-activated cell sorting. This method of mamma- lian cell display was verified by successfully screening ErbB3 scFv candidates. A published scFv control was used to confirm the feasibility of the ErbB3 scFv screening process. Three ErbB3 scFv candidates were produced and they were found to have affinity similar to the published scFv candidate. Thus, the present screening system provided an optimal alternative for rapid acquisition of a novel candi- date scFv sequence to target genes with high affinity in vitro.

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期刊信息
  • 《生物化学与生物物理学报:英文版》
  • 北大核心期刊(2004版)
  • 主管单位:
  • 主办单位:中国科学院上海生物化学研究所
  • 主编:
  • 地址:上海岳阳路319号
  • 邮编:200031
  • 邮箱:abbs@sibs.ac.cn
  • 电话:021-54920956 54920955
  • 国际标准刊号:ISSN:1672-9145
  • 国内统一刊号:ISSN:31-1940/Q
  • 邮发代号:4-210
  • 获奖情况:
  • 国内外数据库收录:
  • 美国化学文摘(网络版),英国农业与生物科学研究中心文摘,荷兰文摘与引文数据库,美国生物医学检索系统,美国剑桥科学文摘,美国科学引文索引(扩展库),美国生物科学数据库,英国动物学记录,日本日本科学技术振兴机构数据库,中国中国科技核心期刊,中国北大核心期刊(2004版),英国英国皇家化学学会文摘,中国北大核心期刊(2000版)
  • 被引量:5851