Journal of Biomolecular Screening

 

Advanced Search

Journal Navigation

Journal Home

Subscriptions

Archive

Contact Us

Table of Contents

Click here for more information

Sign In to gain access to subscriptions and/or personal tools.
This Article
Right arrow Full Text (PDF)
Right arrow All Versions of this Article:
1087057106298538v1
12/2/285    most recent
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 ISI 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 Google Scholar
Google Scholar
Right arrow Articles by Cassutt, K. J.
Right arrow Articles by Tang, W.
Right arrow Search for Related Content
PubMed
Right arrow PubMed Citation
Right arrow Articles by Cassutt, K. J.
Right arrow Articles by Tang, W.
Social Bookmarking
Add to CiteULike   Add to Connotea   Add to Del.icio.us   Add to Digg   Add to Reddit   Add to Technorati  
What's this?
This version was published on March 1, 2007
Journal of Biomolecular Screening, Vol. 12, No. 2, 285-287 (2007)
DOI: 10.1177/1087057106298538
© 2007 Society for Biomolecular Sciences

Identifying Nonselective Hits from a Homogeneous Calcium Assay Screen

Kelly J. Cassutt

Hamamatsu Corp., Bridgewater, NJ

Michael J. Orsini

Mojgan Abousleiman

Dean Colone

Weimin Tang

Pharmaceutical Research and Development, Johnson and Johnson, Raritan, NJ

The authors used a homogeneous calcium dye kit with a cell line transfected using a recombinant protein construct to screen a 50,000 compound library for G-protein coupled receptor (GPCR) agonists. Only 1 of the 365 primary hits activated Gq-coupled GPCRs, as shown using IP-ONE HTRF®. Furthermore, an agonist screen against the entire compound library and same heterologous cell line using AequoScreenTM technology generated no false positives and identified the same positive hit. Next, a multiplex assay composed of both Fluo-3 and Fura-2–loaded cells identified 1 false positive and the same true-positive hit out of the 365 primary hits. Finally, rescreening the 365 primary hits against the parental cell line loaded using the homogeneous calcium dye kit confirmed the specificity of the same true-positive hit only. In summary, the results suggest that AequoScreenTM technology, IP-ONE HTRF®, and multiplex assays are unique, orthogonal technologies to identify nonspecific hits.

Key Words: G-protein-coupled receptor • homogeneous calcium assay • aequorin • functional drug screening system • Fura-2 • Fluo-3 • IP-ONE HTRF®


Add to CiteULike CiteULike   Add to Connotea Connotea   Add to Del.icio.us Del.icio.us   Add to Digg Digg   Add to Reddit Reddit   Add to Technorati Technorati    What's this?