SPM for functional identification of individual biomolecules

Robert Ros; Falk Schwesinger; Celestino Padeste; Andreas Plueckthun; Dario Anselmetti; Hans-Joachim Guentherodt; Louis Tiefenauer

doi:10.1117/12.350621

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17 June 1999 SPM for functional identification of individual biomolecules

Robert Ros, Falk Schwesinger, Celestino Padeste, Andreas Plueckthun, Dario Anselmetti, Hans-Joachim Guentherodt, Louis Tiefenauer

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Proceedings Volume 3607, Scanning and Force Microscopies for Biomedical Applications; (1999) https://doi.org/10.1117/12.350621
Event: BiOS '99 International Biomedical Optics Symposium, 1999, San Jose, CA, United States

Abstract

The identification of specific binding molecules is of increasing interest in the context of drug development based on combinatorial libraries. Scanning Probe Microscopy (SPM) is the method of choice to image and probe individual biomolecules on a surface. Functional identification of biomolecules is a first step towards screening on a single molecule level. As a model system we use recombinant single- chain Fv fragment (scFv) antibody molecules directed against the antigen fluorescein. The scFv's are covalently immobilized on a flat gold surface via the C-terminal cysteine, resulting in a high accessibility of the binding site. The antigen is immobilized covalently via a long hydrophilic spacer to the silicon nitride SPM-tip. This arrangement allows a direct measurement of binding forces. Thus, closely related antibody molecules differing in only one amino acid at their binding site could be distinguished. A novel SPM-software has been developed which combines imaging, force spectroscopic modes, and online analysis. This is a major prerequisite for future screening methods.

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Robert Ros, Falk Schwesinger, Celestino Padeste, Andreas Plueckthun, Dario Anselmetti, Hans-Joachim Guentherodt, and Louis Tiefenauer "SPM for functional identification of individual biomolecules", Proc. SPIE 3607, Scanning and Force Microscopies for Biomedical Applications, (17 June 1999); https://doi.org/10.1117/12.350621

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