Title

Diffusion and binding properties investigated by Fluorescence Correlation Spectroscopy (FCS)

UMMS Affiliation

Department of Biochemistry and Molecular Pharmacology; RNA Therapeutics Institute

Publication Date

2005-10-27

Document Type

Article

Subjects

*Algorithms; Binding Sites; Biopolymers; Diffusion; Fluorescent Antibody Technique; Kinetics; Protein Interaction Mapping; Spectrometry, Fluorescence; Statistics as Topic

Disciplines

Analytical Chemistry | Biochemistry | Biochemistry, Biophysics, and Structural Biology | Biotechnology | Molecular Biology | Pharmacology, Toxicology and Environmental Health

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

DOI of Published Version

10.2174/138920105774370616

Source

Grünwald D, Cardoso MC, Leonhardt H, Buschmann V. Diffusion and binding properties investigated by Fluorescence Correlation Spectroscopy (FCS). Curr Pharm Biotechnol. 2005 Oct;6(5):381-6. doi:10.2174/138920105774370616

Journal/Book/Conference Title

Current pharmaceutical biotechnology

Comments

At the time of publication, David Grünwald was not yet affiliated with the University of Massachusetts Medical School.

Related Resources

Link to Article in PubMed

PubMed ID

16248811

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