Molecular Modeling and Affinity Determination of scFv Antibody: Proper Linker Peptide Enhances Its Activity

	Molecular Modeling and Affinity Determination of scFv Antibody: Proper Linker Peptide Enhances Its Activity
	Gu X; Jia XL(贾潇凌); Feng JN; Shen BF; Huang Y; Geng SS; Sun YX; Wang Y; Li Y; Long M(龙勉)
刊名	Annals of Biomedical Engineering
	2010
通讯作者邮箱	mlong@imech.ac.cn
卷号	38 期号:2 页码:537-549
关键词	Linker peptide Molecular modeling Micropipette adhesion frequency Scatchard analysis Binding affinity
ISSN号	0090-6964
通讯作者	龙勉
合作状况	国内
中文摘要	One of existing strategies to engineer active antibody is to link VH and VL domains via a linker peptide. How the composition, length, and conformation of the linker affect antibody activity, however, remains poorly understood. In this study, a dual approach that coordinates molecule modeling, biological measurements, and affinity evaluation was developed to quantify the binding activity of a novel stable miniaturized anti-CD20 antibody or singlechain fragment variable (scFv) with a linker peptide. Upon computer-guided homology modeling, distance geometry analysis, and molecular superimposition and optimization, three new linker peptides PT1, PT2, and PT3 with respective 7, 10, and 15 residues were proposed and three engineered antibodies were then constructed by linking the cloned VH and VL domains and fusing to a derivative of human IgG1. The binding stability and activity of scFv-Fc chimera to CD20 antigen was quantified using a micropipette adhesion frequency assay and a Scatchard analysis. Our data indicated that the binding affinity was similar for the chimera with PT2 or PT3 and ~24-fold higher than that for the chimera with PT1, supporting theoretical predictions in molecular modeling. These results further the understanding in the impact of linker peptide on antibody structure and activity.
学科主题	生物力学
类目[WOS]	Engineering, Biomedical
研究领域[WOS]	Engineering
关键词[WOS]	LIGAND BINDING-KINETICS ; SINGLE-CHAIN ANTIBODY ; ALKALINE-PHOSPHATASE ; CRYSTAL-STRUCTURE ; ESCHERICHIA-COLI ; FLOW-CYTOMETRY ; FV FRAGMENT ; CELL-CYCLE ; ANTIGEN ; DESIGN
收录类别	SCI
语种	英语
WOS记录号	WOS:000274237000024
公开日期	2009-12-31
内容类型	期刊论文
源URL	[http://dspace.imech.ac.cn/handle/311007/33025]
专题	力学研究所_国家微重力实验室
通讯作者	Long M(龙勉)
推荐引用方式 GB/T 7714	Gu X,Jia XL,Feng JN,et al. Molecular Modeling and Affinity Determination of scFv Antibody: Proper Linker Peptide Enhances Its Activity[J]. Annals of Biomedical Engineering,2010,38(2):537-549.
APA	Gu X.,Jia XL.,Feng JN.,Shen BF.,Huang Y.,...&龙勉.(2010).Molecular Modeling and Affinity Determination of scFv Antibody: Proper Linker Peptide Enhances Its Activity.Annals of Biomedical Engineering,38(2),537-549.
MLA	Gu X,et al."Molecular Modeling and Affinity Determination of scFv Antibody: Proper Linker Peptide Enhances Its Activity".Annals of Biomedical Engineering 38.2(2010):537-549.