Protein chip analysis by probing time-resolved UV fluorescence

Paulius Grigaravicius; Rüdiger Dietrich; Wolfgang Fritzsche; Karl Otto Greulich; Uwe Horn; Dietmar Knoll; Sven Peters; Hans-Martin Striebel; Peter Schellenberg

doi:10.1117/12.728451

13 July 2007 Protein chip analysis by probing time-resolved UV fluorescence

Paulius Grigaravicius, Rüdiger Dietrich, Wolfgang Fritzsche, Karl Otto Greulich, Uwe Horn, Dietmar Knoll, Sven Peters, Hans-Martin Striebel, Peter Schellenberg

Author Affiliations +

Proceedings Volume 6633, Biophotonics 2007: Optics in Life Science; 66332E (2007) https://doi.org/10.1117/12.728451
Event: European Conferences on Biomedical Optics, 2007, Munich, Germany

Abstract

We describe a novel label-free method to analyse protein interactions on microarrays as well as in solution. By this technique the time resolved native protein fluorescence in the UV is probed. The method is based on alterations of the protein upon ligand binding, and, as a consequence, of alterations of the environment of the proteins' aromatic amino acids. These amino acids act as internal probes, and as a result, the fluorescence lifetime of the proteins change due to binding to a ligand partner such as another protein. We were able to demonstrate the feasibility of the method with many compounds, including protein-protein, protein-antibody, protein-nucleic acid and protein-small ligand pairs. Unlike to many other label-free techniques, the sensitivity of the method does not depend on the size of the counterbinding ligand and therefore is particularly suitable for drug monitoring, when small molecules are involved.

Citation Download Citation

Paulius Grigaravicius, Rüdiger Dietrich, Wolfgang Fritzsche, Karl Otto Greulich, Uwe Horn, Dietmar Knoll, Sven Peters, Hans-Martin Striebel, and Peter Schellenberg "Protein chip analysis by probing time-resolved UV fluorescence", Proc. SPIE 6633, Biophotonics 2007: Optics in Life Science, 66332E (13 July 2007); https://doi.org/10.1117/12.728451

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