| What's New |
| Promotions |
| Events/Invites |
Emerging Trends in Utilizing Cell Lines as Reagents – Novel Methods for High Throughput Screening Applications
A review of recent literature devoted to GPCR high throughput screening (HTS) shows an increasing reliance on the use of cell based assays - despite the inherent challenges in their use.
Although now widely accepted, cell-based assays have the advantage that the GPCR is studied in a functional and therefore more physiologically relevant context. Nonetheless, the use of cells in HTS also features several drawbacks in the screening environment, not the least of which are the large numbers required for screening, as well as the significant resource requirements to overcome their complexity.
Bergsdorf, et. al., note in the February 2008 issue of Assay and Drug Development Technologies that "compared to biochemical high-throughput screening (HTS) assays, cell-based functional assays are generally thought to be more time consuming and complex because of additional efforts for running continuous cell cultures as well as the numerous assay steps when transferring media and compounds. However, such systems require substantial investments in sophisticated hardware and highly specialized personnel."
In the June 2008 issue of the Journal of Biomolecular Screening, Gilchrist, et. al. point out that "the pharmaceutical industry must triple the number of new entities screened each year to meet increasing demand for new drug candidates," and further note that, "miniaturized cell-based functional assays have become increasingly important in HTS."
Both authors propose novel solutions for reaping the benefits of utilizing cell lines as reagents, and for overcoming the challenges in doing so. Bergsdorf and colleagues describe an elegant method that overcomes the throughput demands of cells-as-reagents, including:
The use of “cryopreserved cell aliquots instead of continuous cell culture"
Using “cells in suspension instead of adherent cells"
And utilizing “ready-to-screen assay plates with nanoliter aliquots of test compounds"
Their assay methodology translated well into a high-throughput format, and “the assay quality was sufficient to run HTS campaigns" in low and high density formats, “with good Z’ factors and excellent reproducibility of results."
The Gilchrist team’s research utilized PerkinElmer’s “AequoScree CHO-K1 cell line, heterologously expressing 5-HT2b receptor and apoaequorin" to investigate “aequorin-based detection of calcium mobilization as a means for enabling miniaturized functional GPCR HTS campaigns."
They concluded that aequorin assays provide “several advantages over the fluorescence-based methods for HTS of GPCR-induced calcium mobilization." The Gilchrist team was also able to conclude that the aequorin assay “is highly amenable to miniaturization" in high-density formats such as 1536-well, and furthermore, could be miniaturized “while maintaining and/or increasing statistical robustness of agonist and antagonist hit identification assays".
Taken together, current and emerging research continues to identify methods and technologies for enabling HTS programs designed around the concept of cell lines as reagents, with compelling results. New miniaturization approaches for minimizing amounts of needed reagents coupled with novel strategies for maintaining efficacy are becoming more widespread in the research community, providing better screening outcomes and accelerating the identification of novel compound leads.
PerkinElmer has the largest collection of validated cell lines and photoprotein cell lines for your GPCR research and discovery efforts. PerkinElmer offers stable mammalian cell lines over expressing a variety of recombinant GPCRs. Certified mycoplasma-free, they are tested for receptor expression levels, and in some cases, for functional coupling. Validation is done with PerkinElmer complementary membrane preparations and radioligands.
For more information on our complete cell line offering click here and visit our web today!