New method for determination of average scFv fragment number displayed on the M13 phage surface

	New method for determination of average scFv fragment number displayed on the M13 phage surface
	Zhao, Peng 1; Zhu, Guijie 1; Zhang, Lihua 1; Liang, Zhen 1; Li, Zonghai 2; Zhang, Yukui 1
刊名	pure and applied chemistry
	2010
卷号	82 期号:1 页码:205-211
关键词	capillary electrophoresis with laser-induced fluorescence enhanced green fluorescent protein quantitative evaluation single-chain-Fv display M13 phage library
通讯作者	张丽华
产权排序	1;1
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.
WOS标题词	science & technology ; physical sciences
类目[WOS]	chemistry, multidisciplinary
研究领域[WOS]	chemistry
关键词[WOS]	green fluorescent protein ; gene-transfer ; antibodies ; libraries ; binding ; cells ; site
收录类别	SCI
原文出处	false
语种	英语
WOS记录号	WOS:000274709300018
公开日期	2010-11-30
内容类型	期刊论文
源URL	[http://159.226.238.44/handle/321008/103203]
专题	大连化学物理研究所_中国科学院大连化学物理研究所
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
推荐引用方式 GB/T 7714	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.
APA	Zhao, Peng,Zhu, Guijie,Zhang, Lihua,Liang, Zhen,Li, Zonghai,&Zhang, Yukui.(2010).New method for determination of average scFv fragment number displayed on the M13 phage surface.pure and applied chemistry,82(1),205-211.
MLA	Zhao, Peng,et al."New method for determination of average scFv fragment number displayed on the M13 phage surface".pure and applied chemistry 82.1(2010):205-211.