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Bibliographic Metadata
- TitleAntigen binding forces of individually addressed single-chain Fv antibody molecules
- Author
- Is part ofProceedings of the National Academy of Sciences of the United States of America, Vol. 95 Issue 13, page 7402-7405
- Published
- LanguageEnglish
- Document typeJournal Article
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Abstract
Antibody single-chain Fv fragment (scFv) molecules that are specific for fluorescein have been engineered with a C-terminal cysteine for a directed immobilization on a flat gold surface. Individual scFv molecules can be identified by atomic force microscopy. For selected molecules the antigen binding forces are then determined by using a tip modified with covalently immobilized antigen. An scFv mutant of 12 percent lower free energy for ligand binding exhibits a statistically significant 20 percent lower binding force. This strategy of covalent immobilization and measuring well separated single molecules allows the characterization of ligand binding forces in molecular repertoires at the single molecule level and will provide a deeper insight into biorecognition processes.
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