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Title: Molecular Modeling and Affinity Determination of scFv Antibody: Proper Linker Peptide Enhances Its Activity
Author: 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)
Source: Annals of Biomedical Engineering
Issued Date: 2010
Volume: 38, Issue:2, Pages:537-549
Abstract: 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.
Keyword: Linker peptide ; Molecular modeling ; Micropipette adhesion frequency ; Scatchard analysis ; Binding affinity
Language: 英语
Indexed Type: SCI
Corresponding Author: 龙勉
Correspondent Email: mlong@imech.ac.cn
DOI: 10.1007/s10439-009-9810-2
DOC Type: Article
WOS Subject: Engineering, Biomedical
WOS Subject Extended: Engineering
WOS Keyword Plus: LIGAND BINDING-KINETICS ; SINGLE-CHAIN ANTIBODY ; ALKALINE-PHOSPHATASE ; CRYSTAL-STRUCTURE ; ESCHERICHIA-COLI ; FLOW-CYTOMETRY ; FV FRAGMENT ; CELL-CYCLE ; ANTIGEN ; DESIGN
WOS ID: WOS:000274237000024
ISSN: 0090-6964
Subject: 交叉与边缘领域的力学::生物力学
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Content Type: 期刊论文
URI: http://dspace.imech.ac.cn/handle/311007/33025
Appears in Collections:国家微重力实验室_期刊论文

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