Current Pharmaceutical Biotechnology

Zeno Foldes-Papp
Visiting Professor of Medical Biochemistry
HELIOS Clinical Center of Emergency Medicine
Department for Internal Medicine
Alte-Koelner-Strasse 9
D-51688 Koeln-Wipperfuerth
Germany

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Diffusion and Binding Properties Investigated by Fluorescence Correlation Spectroscopy (FCS)

Author(s): D. Grunwald, M. C. Cardoso, H. Leonhardt, V. Buschmann.

Abstract:

During the last years, Fluorescence Correlation Spectroscopy (FCS) has proven to be a powerful tool for basic research in many applications. The combination of a minimal detection volume in the femtoliter range coupled with very high sensitivity extends the possibilities to design sensitive homogeneous tests. In this article we illustrate the analysis of binding processes with FCS based on the changes in diffusion characteristics of GFP upon binding to an antibody. Problems induced by highly heterogeneous samples are discussed and differences of GFP binding to a monoclonal and a polyclonal antibody are shown and analyzed. We stress data processing, limitations and useful approximations in FCS methodology. Basic ideas of data acquisition and processing as well as new developments and applications are presented.

Keywords: fluorescence correlation spectroscopy, gfp, kinetics, binding assay, antibody

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Article Details

VOLUME: 6
ISSUE: 5
Year: 2005
Page: [381 - 386]
Pages: 6
DOI: 10.2174/138920105774370616
Price: $58