中国科学院大连化学物理研究所机构知识库
Advanced  
DICP OpenIR  > 中国科学院大连化学物理研究所  > 期刊论文
题名: New method for determination of average scFv fragment number displayed on the M13 phage surface
作者: Zhao, Peng1;  Zhu, Guijie1;  Zhang, Lihua1;  Liang, Zhen1;  Li, Zonghai2;  Zhang, Yukui1
通讯作者: 张丽华
关键词: capillary electrophoresis with laser-induced fluorescence ;  enhanced green fluorescent protein ;  quantitative evaluation ;  single-chain-Fv display M13 phage library
刊名: PURE AND APPLIED CHEMISTRY
发表日期: 2010
DOI: 10.1351/PAC-CON-09-02-03
卷: 82, 期:1, 页:205-211
收录类别: SCI
文章类型: Article
部门归属: 18
项目归属: 1810
产权排名: 1;1
WOS标题词: Science & Technology ;  Physical Sciences
类目[WOS]: Chemistry, Multidisciplinary
研究领域[WOS]: Chemistry
英文摘要: Single-chain-Fv (scFv) display M13 phage library has been regarded as a powerful tool for screening specific antibodies via binding with target proteins. Generally the library quality is evaluated through detecting gene fragments by molecular biology methods, which is not only time- and labor-consuming, but also impossible to obtain quantitative information about the binding capacity of the phage library. In our recent study, a new method to calculate the average scFv number displayed on the M13 phage surface was proposed by capillary electrophoresis with laser-induced fluorescence (CE-LIF) detection. By this method, enhanced green fluorescent protein (EGFP) and scFv phage clones that could specifically bind with EGFP were mixed with different ratios, followed by analysis by CE-LIF. With the dilution of EGFP by phage solution, the peak areas of scFv phage clones and free EGFP were decreased continuously, while that of the EGFP-M13 phage complex was found to decrease initially, then trend to be stable, and finally decrease further. When the volume ratio of the M13 phage to EGFP reached 660:1, corresponding to the molecule number ratio as 1:2.6, no more EGFP was found to bind with the M13 phage, which demonstrated that, by average, 2.6 scFv fragments that could bind with EGFP were displayed on the M13 phage surface. All these experimental results demonstrated that, by such a method, the quantitative evaluation of the phage library could be achieved with high throughput and accuracy.
关键词[WOS]: GREEN FLUORESCENT PROTEIN ;  GENE-TRANSFER ;  ANTIBODIES ;  LIBRARIES ;  BINDING ;  CELLS ;  SITE
语种: 英语
原文出处: 查看原文
WOS记录号: WOS:000274709300018
Citation statistics: 
内容类型: 期刊论文
URI标识: http://cas-ir.dicp.ac.cn/handle/321008/103203
Appears in Collections:中国科学院大连化学物理研究所_期刊论文

Files in This Item:

There are no files associated with this item.


作者单位: 1.Chinese Acad Sci, Key Lab Separat Sci Analyt Chem, Natl Chromatog R&A Ctr, Dalian Inst Chem Phys, Dalian 116023, Peoples R China
2.Shanghai Jiao Tong Univ, Shanghai Canc Inst, Shanghai 200240, Peoples R China

Recommended Citation:
Zhao, Peng,Zhu, Guijie,Zhang, Lihua,et al. New method for determination of average scFv fragment number displayed on the M13 phage surface[J]. PURE AND APPLIED CHEMISTRY,2010,82(1):205-211.
Service
 Recommend this item
 Sava as my favorate item
 Show this item's statistics
 Export Endnote File
Google Scholar
 Similar articles in Google Scholar
 [Zhao, Peng]'s Articles
 [Zhu, Guijie]'s Articles
 [Zhang, Lihua]'s Articles
CSDL cross search
 Similar articles in CSDL Cross Search
 [Zhao, Peng]‘s Articles
 [Zhu, Guijie]‘s Articles
 [Zhang, Lihua]‘s Articles
Related Copyright Policies
Null
Social Bookmarking
  Add to CiteULike  Add to Connotea  Add to Del.icio.us  Add to Digg  Add to Reddit 
所有评论 (0)
暂无评论
 
评注功能仅针对注册用户开放,请您登录
您对该条目有什么异议,请填写以下表单,管理员会尽快联系您。
内 容:
Email:  *
单位:
验证码:   刷新
您在IR的使用过程中有什么好的想法或者建议可以反馈给我们。
标 题:
 *
内 容:
Email:  *
验证码:   刷新

Items in IR are protected by copyright, with all rights reserved, unless otherwise indicated.

 

 

Valid XHTML 1.0!
Powered by CSpace