SPM for functional identification of individual biomolecules

Robert Ros, Falk Schwesinger, Celestino Padeste, Andreas Pluckthun, Dario Anselmetti, Hans Joachim Guntherodt, Louis Tiefenauer

Research output: Chapter in Book/Report/Conference proceedingChapter

5 Scopus citations

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.

Original languageEnglish (US)
Title of host publicationProceedings of SPIE - The International Society for Optical Engineering
PublisherSociety of Photo-Optical Instrumentation Engineers
Pages84-89
Number of pages6
Volume3607
StatePublished - 1999
Externally publishedYes
EventProceedings of the 1999 Scanning and Force Microscopies for Biomedical Applications - San Jose, CA, USA
Duration: Jan 24 1999Jan 25 1999

Other

OtherProceedings of the 1999 Scanning and Force Microscopies for Biomedical Applications
CitySan Jose, CA, USA
Period1/24/991/25/99

ASJC Scopus subject areas

  • Electrical and Electronic Engineering
  • Condensed Matter Physics

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