Advanced Search

Journal Navigation

Journal Home

Subscriptions

Archive

Contact Us

Table of Contents

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
Right arrow Citation Map
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 (25)
Right arrow Citing Articles via Google Scholar
Right arrow Citing Articles via Scopus
Google Scholar
Right arrow Articles by Turconi, S.
Right arrow Articles by Pope, A. J.
Right arrow Search for Related Content
PubMed
Right arrow PubMed Citation
Right arrow Articles by Turconi, S.
Right arrow Articles by Pope, A. 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?

Real Experiences of uHTS: A Prototypic 1536-Well Fluorescence Anisotropy-Based uHTS Screen and Application of Well-Level Quality Control Procedures

Sandra Turconi

Molecular Interactions & New Assay Technologies, GlaxoSmithKline Pharmaceuticals, Harlow, Essex, UK

Kerry Shea

Molecular Interactions & New Assay Technologies, GlaxoSmithKline Pharmaceuticals, Harlow, Essex, UK

Stephen Ashman

Molecular Interactions & New Assay Technologies, GlaxoSmithKline Pharmaceuticals, Harlow, Essex, UK

Kenneth Fantom

Molecular Interactions & New Assay Technologies, GlaxoSmithKline Pharmaceuticals, Harlow, Essex, UK

David L. Earnshaw

Molecular Interactions & New Assay Technologies, GlaxoSmithKline Pharmaceuticals, Harlow, Essex, UK

Ryan P. Bingham

Molecular Interactions & New Assay Technologies, GlaxoSmithKline Pharmaceuticals, Harlow, Essex, UK

Ulrich M. Haupts

Molecular Interactions & New Assay Technologies, GlaxoSmithKline Pharmaceuticals, Harlow, Essex, UK

Murray J.B. Brown

Molecular Interactions & New Assay Technologies, GlaxoSmithKline Pharmaceuticals, Harlow, Essex, UK

Andrew J. Pope

Molecular Interactions & New Assay Technologies, GlaxoSmithKline Pharmaceuticals, Harlow, Essex, UK

This paper describes, for the first time, a true ultra-high throughput screen (uHTS) based upon fluorescence anisotropy and performed entirely in 1536-well assay plates. The assay is based upon binding and displacement of a BODIPY-FL-labeled antibiotic to a specific binding site on 70S ribosomes from Escherichia coli (Kd 15 nM). The screen was performed at uHTS rates (i.e., >100,000 assay wells/24 h) using entirely commercially available equipment. In order to examine the reproducibility of detection of test compound effects, assays were performed in duplicate. Both overall assay statistics and reproducibility for individual compound results were excellent, at least equivalent to conventional HTS assays. Interference artifacts occurred mainly as a result of autofluorescence from test compounds. Well-level quality control procedures were developed to detect, eliminate, or even correct for such effects. Moreover, development of a brighter, longer wavelength probe (based upon Cy3B) markedly reduced such interferences. Overall, the data demonstrate that fluorescence anisotropy-based uHTS is now a practical reality.

Journal of Biomolecular Screening, Vol. 6, No. 5, 275-290 (2001)
DOI: 10.1177/108705710100600502
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
 Antimicrob. Agents Chemother.Home page
K. Yan, E. Hunt, J. Berge, E. May, R. A. Copeland, and R. R. Gontarek
Fluorescence Polarization Method To Characterize Macrolide-Ribosome Interactions
Antimicrob. Agents Chemother., August 1, 2005; 49(8): 3367 - 3372.
[Abstract] [Full Text] [PDF]

Home page
 Nucleic Acids ResHome page
T. Göhler, S. Jäger, G. Warnecke, H. Yasuda, E. Kim, and W. Deppert
Mutant p53 proteins bind DNA in a DNA structure-selective mode
Nucleic Acids Res., February 18, 2005; 33(3): 1087 - 1100.
[Abstract] [Full Text] [PDF]

Home page
 J Biomol ScreenHome page
P. Blommel, G. T. Hanson, and K. W. Vogel
Multiplexing Fluorescence Polarization Assays to Increase Information Content Per Screen: Applications for Screening Steroid Hormone Receptors
J Biomol Screen, June 1, 2004; 9(4): 294 - 302.
[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
A. Harris, S. Cox, D. Burns, and C. Norey
Miniaturization of Fluorescence Polarization Receptor-Binding Assays Using CyDye-Labeled Ligands
J Biomol Screen, August 1, 2003; 8(4): 410 - 420.
[Abstract] [PDF]

Home page
 J Biomol ScreenHome page
T. C. Turek-Etienne, E. C. Small, S. C. Soh, T. A. Xin, P. V. Gaitonde, E. B. Barrabee, R. F. Hart, and R. W. Bryant
Evaluation of Fluorescent Compound Interference in 4 Fluorescence Polarization Assays: 2 Kinases, 1 Protease, and 1 Phosphatase
J Biomol Screen, April 1, 2003; 8(2): 176 - 184.
[Abstract] [PDF]