Join us for an informative webinar, discussing the use of cloned cells for research. It will include presentations from the Scientific Director at Amgen, Jeff Reagan; the Co-Founder and EVP of Molecular Sensing, Inc., Scot Weinberger; and the Director of Cell & Molecular Biology at Chantest, Luke Armstrong. Evaluation of the Mechanism of Action of Allosteric Modulators and its Application to Drug Discovery Allosteric modulators are a class of drugs which bind to a site distinct from the orthosteric binding site on a receptor or enzyme. These types of molecules offer potential benefits in terms of both safety and efficacy for the treatment of various diseases. Calcimimetics are positive allosteric modulators at the calcium sensing receptor. In this lecture, we will discuss the use of calcimimetics for the treatment of secondary hyperparathyroidism. This presentation will discuss the application of an operational model of allosteric agonism/modulation to aid in understanding the mechanism of action of calcimimetics. In addition, an emerging technology based upon backscattering interferometry will be discussed. This technology allows one to determine both the affinity and cooperativity of allosteric modulators to a receptor using a label-free system. The information derived from this assay will allow one to couple it to information obtained from cell-based assays, in order to gain a better understanding of the mechanism of action of allosteric modulators. Label-Free, Free-Solution Assays in the Study of Orthosteric and Allosteric GPCR Ligand Binding Using Back-Scattering Interferometry Back-scattering interferometry (BSI) is an emerging label-free technology for mass- and matrix-independent biophysical characterization of small molecule interaction with complex integral membrane proteins in a native-like environment. Integral membrane proteins such as GPCRs are critical targets for drug discovery, but present a host of challenges to the investigation of their biophysical properties. Of paramount interest to drug discovery efforts is the characterization of the interaction of GPCRs with small molecule compounds as a component of library screening, MOA determination, drug candidate profiling, and other applications. The difficulty associated with obtaining target-enriched material that is functionally intact, effectively restricts the range of assay techniques suited to quantifying compound affinity data in vitro. Fluorescent and radiolabeled ligands serve a vital role, but are typically employed for only indirect characterization of orthosteric and allosteric ligand binding. BSI allows for the direct determination of compound affinity (Kd), directly for GPCR ligands, without the use of labels or tethering the target to a surface, which is difficult for integral membrane targets and can result in low sensitivity and unwanted artifacts. In this webinar, we will describe the application of BSI to the assay of small molecule ligand binding to human M4 acetylcholine receptor overexpressed in crude membrane fractions in free solution. Both orthosteric and allosteric ligand binding to M4, as well as allostery for this system, will be reviewed and discussed. Dialing for Targets: Procuring Cells Expressing Channels and Receptors for Biotech Research Drug discovery and safety testing for ion channels and GPCRs rely heavily on cell lines expressing recombinant targets, which are assayed for target activity on automated platforms. ChanTest is the leading provider of ion channel safety and discovery services, and also offers the most extensive portfolio of high quality ion channel-expressing cell lines. ChanTest also provides services to develop new ion channel cell lines that are thoroughly validated for functional assay applications (e.g., automated patch clamp and fluorescent assays) . Examples of projects to develop cell lines stably or transiently expressing novel targets for use in automated electrophysiology assays will be presented. For customers who already have cell lines, but are experiencing constraints in making cell banks, ChanTest also offers services for large-scale growth and cryopreservation of customer-provided cell lines. Additionally, ChanTest has capabilities for making membrane preparations from any cell line, and performing radioligand binding assays on the preparations for quality testing. In a partnership with Molecular Sensing, ChanTest membrane preparations are proving to be effective for analysis by backscattering interferometry. In sum, for biotech companies wanting to contract out ion channel cell line projects of any scope, ChanTest is an ideal partner.

• published: 27 May 2014
• views: 3