Advanced Search

Journal Navigation

Journal Home

Subscriptions

Archive

Contact Us

Table of Contents

Click here for more information

CiteULike is a free service for managing and discovering scholarly references - click here to get started.

Sign In to gain access to subscriptions and/or personal tools.
Journal of Biomolecular Screening
This Article
Right arrow Full Text (PDF)
Right arrow References
Right arrow Alert me when this article is cited
Right arrow Alert me if a correction is posted
Services
Right arrow Email this article to a friend
Right arrow Similar articles in this journal
Right arrow Similar articles in Web of Science
Right arrow Similar articles in PubMed
Right arrow Alert me to new issues of the journal
Right arrow Add to Saved Citations
Right arrow Download to citation manager
Right arrowRequest Permissions
Right arrow Request Reprints
Right arrow Add to My Marked Citations
Citing Articles
Right arrow Citing Articles via HighWire
Right arrow Citing Articles via Web of Science (21)
Right arrow Citing Articles via Google Scholar
Right arrow Citing Articles via Scopus
Google Scholar
Right arrow Articles by Scheel, A. A.
Right arrow Articles by Lohse, M. J.
Right arrow Search for Related Content
PubMed
Right arrow PubMed Citation
Right arrow Articles by Scheel, A. A.
Right arrow Articles by Lohse, M. J.
Social Bookmarking
Add to CiteULike   Add to Complore   Add to Connotea   Add to Del.icio.us   Add to Digg   Add to Reddit   Add to Technorati   Add to Twitter  
What's this?

Receptor-Ligand Interactions Studied with Homogeneous Fluorescence-Based Assays Suitable for Miniaturized Screening

Andreas A. Scheel

EVOTEC BioSystems AG, Hamburg, Germany

Bettina Funsch

EVOTEC BioSystems AG, Hamburg, Germany

Michael Busch

EVOTEC BioSystems AG, Hamburg, Germany

Gabriele Gradl

EVOTEC BioSystems AG, Hamburg, Germany

Johannes Pschorr

EVOTEC BioSystems AG, Hamburg, Germany

Martin J. Lohse

Lnstitut fur Pharmakologie, Universitdt Wurzburg, Wurzburg, Germany

Cell membrane receptors play a central role in controlling cellular functions, making them the target of drugs for a wide variety of diseases. This report describes how a recently developed method, fluorescence intensity distribution analysis (FIDA), can be used to develop homogeneous, nonradioactive high throughput screening assays for membrane receptors. With FIDA, free ligand and ligand accumulated on receptor-bearing membrane vesicles can be distinguished on the basis of their particle brightness. This allows the concentration of both bound and free ligand to be determined reliably from a single measurement, without any separation. We demonstrate that ligand affinity, receptor expression level, and potency of inhibitors can be determined using the epidermal growth factor and ß2-adrenergic receptors as model systems. Highly focused confocal optics enable single-molecule sensitivity, and sample volumes can thus be reduced to 1,IL without affecting the quality of the fluorescence signal. Our results demonstrate that FIDA is an ideal method for membrane receptor assays offering substantial benefits for assay development and high throughput pharmaceutical screening.

Journal of Biomolecular Screening, Vol. 6, No. 1, 11-18 (2001)
DOI: 10.1177/108705710100600103
Add to CiteULike CiteULike   Add to Complore Complore   Add to Connotea Connotea   Add to Del.icio.us Del.icio.us   Add to Digg Digg   Add to Reddit Reddit   Add to Technorati Technorati   Add to Twitter Twitter    What's this?


This article has been cited by other articles:


Home page
 J Biomol ScreenHome page
H. Harma, A. Rozwandowicz-Jansen, E. Martikkala, H. Frang, I. Hemmila, N. Sahlberg, V. Fey, M. Perala, and P. Hanninen
A New Simple Cell-Based Homogeneous Time-Resolved Fluorescence QRET Technique for Receptor-Ligand Interaction Screening
J Biomol Screen, September 1, 2009; 14(8): 936 - 943.
[Abstract] [PDF]

Home page
 J Biomol ScreenHome page
W. R. Leifert, A. L. Aloia, O. Bucco, R. V. Glatz, and E. J. McMurchie
G-Protein-Coupled Receptors in Drug Discovery: Nanosizing Using Cell-Free Technologies and Molecular Biology Approaches
J Biomol Screen, December 1, 2005; 10(8): 765 - 779.
[Abstract] [PDF]

Home page
 J Biomol ScreenHome page
N. Benson, H. F. Boyd, J. R. Everett, J. Fries, P. Gribbon, N. Haque, K. Henco, T. Jessen, W. H. Martin, T. J. Mathewson, et al.
NanoStore: A Concept for Logistical Improvements of Compound Handling in High-Throughput Screening
J Biomol Screen, September 1, 2005; 10(6): 573 - 580.
[Abstract] [PDF]

Home page
 J Biomol ScreenHome page
S. Jager, N. Garbow, A. Kirsch, H. Preckel, F. U. Gandenberger, K. Herrenknecht, M. Rudiger, J. P. Hutchinson, R. P. Bingham, F. Ramon, et al.
A Modular, Fully Integrated Ultra-High-Throughput Screening System Based on Confocal Fluorescence Analysis Techniques
J Biomol Screen, December 1, 2003; 8(6): 648 - 659.
[Abstract] [PDF]

Home page
 J Biomol ScreenHome page
U. Haupts, M. Rudiger, S. Ashman, S. Turconi, R. Bingham, C. Wharton, J. Hutchinson, C. Carey, K. J. Moore, and A. J. Pope
Single-Molecule Detection Technologies in Miniaturized High-Throughput Screening: Fluorescence Intensity Distribution Analysis
J Biomol Screen, January 1, 2003; 8(1): 19 - 33.
[Abstract] [PDF]

Home page
 J Biomol ScreenHome page
M. Klumpp, A. Scheel, E. Lopez-Calle, M. Busch, K. J. Murray, and A. J. Pope
Ligand Binding to Transmembrane Receptors on Intact Cells or Membrane Vesicles Measured in a Homogeneous 1-Microliter Assay Format
J Biomol Screen, June 1, 2001; 6(3): 159 - 170.
[Abstract] [PDF]

Home page
 J Biomol ScreenHome page
M. Rudiger, U. Haupts, K. J. Moore, and A. J. Pope
Single-Molecule Detection Technologies in Miniaturized High Throughput Screening: Binding Assays for G Protein-Coupled Receptors Using Fluorescence Intensity Distribution Analysis and Fluorescence Anisotropy
J Biomol Screen, February 1, 2001; 6(1): 29 - 37.
[Abstract] [PDF]