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

doi:10.1007/s10439-009-9810-2

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	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); Long M(龙勉)
发表期刊	Annals of Biomedical Engineering
	2010
卷号	38 期号:2 页码:537-549
ISSN	0090-6964
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.
关键词	Linker Peptide Molecular Modeling Micropipette Adhesion Frequency Scatchard Analysis Binding Affinity
学科领域	生物力学
DOI	10.1007/s10439-009-9810-2
收录类别	SCI
语种	英语
WOS记录号	WOS:000274237000024
关键词[WOS]	LIGAND BINDING-KINETICS ; SINGLE-CHAIN ANTIBODY ; ALKALINE-PHOSPHATASE ; CRYSTAL-STRUCTURE ; ESCHERICHIA-COLI ; FLOW-CYTOMETRY ; FV FRAGMENT ; CELL-CYCLE ; ANTIGEN ; DESIGN
WOS研究方向	Engineering
WOS类目	Engineering, Biomedical
引用统计	被引频次：30[WOS] [WOS记录] [WOS相关记录]
文献类型	期刊论文
条目标识符	http://dspace.imech.ac.cn/handle/311007/33025
专题	微重力重点实验室
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.

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