Antigen binding forces of individually addressed single-chain Fv antibody molecules

Robert Ros, Falk Schwesinger, Dario Anselmetti, Martina Kubon, Rolf Schäfer, Andreas Plückthun, Louis Tiefenauer

Research output: Contribution to journalArticle

229 Scopus citations

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% lower free energy for ligand binding exhibits a statistically significant 20% 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.

Original languageEnglish (US)
Pages (from-to)7402-7405
Number of pages4
JournalProceedings of the National Academy of Sciences of the United States of America
Volume95
Issue number13
DOIs
StatePublished - Jun 23 1998

ASJC Scopus subject areas

  • General

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