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Thank you for your interest in MDS Analytical Technologies' CellKey™ System. Documents marked with an asterisk (*) contain customer approved collaboration data or new research data, thus require a simple, one-time registration. Or, if you have already registered, you must be logged in.
Poster
New Applications |
(organized in reverse chronological order) |
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 |  | | | Introducing the CellKey™384 System, a 384-well label-free cellular assay platform for biorelevant drug discovery applications. In this poster we demonstrate the ease of scale up from 96-well to 384-well assay format without compromising the quality of output data. Testing target receptors on several cell lines, including suspension cells, we effectively transferred assay workflows, assay protocols and data analysis parameters to the high throughput system. Consistency of data in terms of CellKey™ response profiles, and EC 50/IC 50 values are demonstrated across both platforms. The data featured here show that the CellKey™384 System enables simple and fast investigation of targets in a high throughput biorelevant drug discovery paradigm. SBS 2008 (Society of Biomolecular Sciences Conference) |  | | | | | Here we successfully demonstrate the feasibility of scaling a label free impedance assay to high throughput screening. Many aspects of assay design, set up and execution are considered, including workflow and measurement duration, while leveraging the inherent ease of use of a label-free technology. We report assay performance statistics including Z’ for multiple plates over multiple days. The CellKey™384 System brings the simplified workflow of a label free assay, the biorelevance of endogenous receptor analysis and the rich information of integrated cellular responses to high throughput screening. SBS 2008 (Society of Biomolecular Sciences Conference) | | | | | | Primary cells provide the most biologically relevant context to study endogenous receptors; however their use early in drug discovery can be prohibitively expensive. The CellKey™ Small Sample 96W microplate enables pharmacological analysis of endogenous receptors at a significantly reduced cell number per well without sacrificing assay reproducibility or robustness. The data presented describe experiments performed using U937 and THP-1 cells, and PBMCs. Experiments were run in parallel on the CellKey™ Standard 96W microplates and the Small Sample 96W microplates. The new Small Sample microplate allows the CellKey™ System to be utilized more cost-effectively with primary cells, facilitating identification of unique and higher-quality compounds efficiently in a label-free biorelevant live-cell assay. Presented at ACT 2007 (IBC's Assay and Cellular Targets Conference) | |
Receptor Panning |
(organized in reverse chronological order) |
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| | | | This poster describes use of the CellKey™ system for receptor identification, characterization, and pharmacology studies. The ability to quickly and easily determine the profile of active receptors in any given cell may be useful in a variety of fields ranging from research to clinical diagnostics. Receptor panning of multiple cell types including CHO, HEK293, HeLa, U- 937, U-2 OS, and TE671 resulted in the identification of many functionally active, differently-coupled endogenous GPCRs, some of which have not been previously documented in the literature. Detailed pharmacological analyses were also performed, including the pharmacological subtyping of GPCRs and Schild analysis. Presented at SBS 2006 (Society of Biomolecular Sciences Conference) | | | | | | The CellKey™ system’s versatility is demonstrated in this analysis of U-2 OS cells. Receptor panning of U-2 OS cells yielded identification of numerous functionally active and differently-coupled endogenous GPCRs, many not previously documented in the literature for these cells. The CellKey™ system also enabled secondary screening and target validation, as shown by an example of histamine receptor subtype identification. Detection and deciphering of cross-talk between GPCR-mediated pathways was also performed. Finally, two different agonists targeting the same receptor generated distinct CDS response profiles, and were then shown to be functionally different by their differential sensitivity to a biochemical modulator. Presented at DDT 2006 (Drug Discovery Technology Conference) | | | | | | CDS was used to measure the functional activity of a comprehensive panel of endogenous G-protein coupled, protein tyrosine kinase, and nuclear receptors in a variety of mammalian cells. Afterward, a subset of active GPCR receptors in the presence and absence of biochemical modulators was studied to deconstruct their observed CDS response and to assess their functional G-protein (Gq, Gi/o, Gs) coupling mechanism. Finally, selective agonists and antagonists were used to confirm the receptor subtype indicated by the unique CDS kinetic response profile. Presented at SBS 2005 (Society of Biomolecular Sciences Conference) | | | | | | This poster demonstrates that CDS can be used to measure mast cell degranulation. The CellKey™ system measured the EC 50 value of Fc eRI stimulation. Antagonist assays were performed using cell-permeable inhibitors of kinases that are involved in the Fc eRI signaling cascade, including Lyn-, PI3-, and Syk-kinases. In addition, the activation of multiple endogenous receptors in mast cells demonstrates the use of CDS in identifying new receptor targets. Presented at SBS 2005 (Society of Biomolecular Sciences Conference), and IIR Cell-Based Assay 2006 Conference | |
Signal Pathway Identification and Deconvolution |
(organized in reverse chronological order) |
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| | | | Real-time characterization of receptors in their native environment is desirable in drug discovery. This poster compares endogenous receptor pharmacology to that of a transfected and over-expressed receptor, and investigates the correlation between these systems. The studies detail the CellKey™ system’s ability to generate robust and reproducible pharmacology data on a panel of agonists (including an inverse agonist) to an important GPCR target expressed in both endogenous and transfected settings. The CellKey™ system’s unique ability to simultaneously provide information about the GPCR pathway activated and the pharmacology of a given ligand is also discussed. Results were compared to conventional assays such as GTP's. Presented at SBS 2006 (Society of Biomolecular Sciences Conference) | | | | | | The CellKey™ system’s versatility is demonstrated in this analysis of U-2 OS cells. Receptor panning of U-2 OS cells yielded identification of numerous functionally active and differently-coupled endogenous GPCRs, many not previously documented in the literature for these cells. The CellKey™ system also enabled secondary screening and target validation, as shown by an example of histamine receptor subtype identification. Detection and deciphering of cross-talk between GPCR-mediated pathways was also performed. Finally, two different agonists targeting the same receptor generated distinct CDS response profiles, and were then shown to be functionally different by their differential sensitivity to a biochemical modulator. Presented at DDT 2006 (Drug Discovery Technology Conference) | | | | | | The identification of live-cell permeable inhibitors for protein kinases is of great interest in therapeutic oncology. This poster shows the epidermal growth factor response in several cancer cell lines and the blocking of the signaling cascades by small-molecule tyrosine kinase inhibitors. The results demonstrate applications of CDS for cell-based compound screening and for the analysis of receptor-ligand signal transduction pathways. Presented at SBS 2005 (Society of Biomolecular Sciences Conference) | | | | | | CDS was used to measure the functional activity of a comprehensive panel of endogenous G-protein coupled, protein tyrosine kinase, and nuclear receptors in a variety of mammalian cells. Afterward, a subset of active GPCR receptors in the presence and absence of biochemical modulators was studied to deconstruct their observed CDS response and to assess their functional G-protein (Gq, Gi/o, Gs) coupling mechanism. Finally, selective agonists and antagonists were used to confirm the receptor subtype indicated by the unique CDS kinetic response profile. Presented at SBS 2005 (Society of Biomolecular Sciences Conference) | | | | | | CDS was used to investigate and compare human MCHR1 signaling in stably transfected CHO and U2-OS cells. MCHR1 transduces its signal via both Gi and Gq G-proteins in transfected cells. The CDS platform enabled the dose-dependent differentiation of Gi- and Gq-coupled events simultaneously in the same microplate well and allowed pharmacological analysis of agonist and antagonist compounds. Further, CDS response profiles demonstrated the complex signal transduction through MCHR1 in a label-free fashion. All results were compared to conventional secondary messenger assays. Presented at SBS 2005 (Society of Biomolecular Sciences Conference), IIR Cell-Based Assay 2006 Conference and IBC ACT 2005 (Assays & Cellular Targets Conference) | | | | | | CDS was used to conduct a pharmacological evaluation of the activation of an endogenously expressed GPCR in two related rhabdomyosarcoma cell lines. CDS was able to consistently measure activation of the endogenous receptor. In contrast, Ca2+ assays required the development of a clonal cell line and the use of non-physiological concentrations of agonist to elicit a response. Presented at SBS 2005 (Society of Biomolecular Sciences Conference), Screentech 2006 and IIR GPCR 2006 Conference | | | | | | This poster presents CDS response profiles that are characteristic of the main subsets of GPCRs within a cell line. Receptor panning was used to investigate the endogenous receptors active in several cell lines: U-2 OS (osteosarcoma), TE-671 (rhabdomyosarcoma) and HeLa (cervical carcinoma). Evaluation of the signaling pathways of three of the hits (histamine, UK14,304 and EGF) was conducted. Presented at DDT 2005 (Drug Discovery Technology Conference) | |
Pharmacological Analysis |
(organized in reverse chronological order) |
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| | | | Cell-based assays are an important aspect of the drug discovery process. However, few investigators appreciate the variability
inherent in routine cell culture practices which can affect assay reproducibility and robustness. Using the CellKey™ system, a labelfree cellular assay technology, we investigated specific cell culture variables, and measured their effects on cellular responses to certain drugs. To quantify cell culture conditions such as cell density and morphology, we used the IncuCyte™ imaging system developed by Essen Instruments. IncuCyte™ is a compact, automated imaging platform designed to provide kinetic, non-invasive live cell imaging. Using these two systems, we were able to document how cell culture conditions can affect cell-based assay data both quantitatively and qualitatively. Our results demonstrate that cellular responses via activated GPCR’s vary with culture conditions and that explicitly monitoring certain cell culture conditions can significantly improve the quality of the data obtained from cell-based assays. Presented at ACT 2006 | | | | | | Real-time characterization of receptors in their native environment is desirable in drug discovery. This poster compares endogenous receptor pharmacology to that of a transfected and over-expressed receptor, and investigates the correlation between these systems. The studies detail the CellKey™ system’s ability to generate robust and reproducible pharmacology data on a panel of agonists (including an inverse agonist) to an important GPCR target expressed in both endogenous and transfected settings. The CellKey™ system’s unique ability to simultaneously provide information about the GPCR pathway activated and the pharmacology of a given ligand is also discussed. Results were compared to conventional assays such as GTP's. Presented at SBS 2006 (Society of Biomolecular Sciences Conference) | | | | | | This poster describes use of the CellKey™ system for receptor identification, characterization, and pharmacology studies. The ability to quickly and easily determine the profile of active receptors in any given cell may be useful in a variety of fields ranging from research to clinical diagnostics. Receptor panning of multiple cell types including CHO, HEK293, HeLa, U- 937, U-2 OS, and TE671 resulted in the identification of many functionally active, differently-coupled endogenous GPCRs, some of which have not been previously documented in the literature. Detailed pharmacological analyses were also performed, including the pharmacological subtyping of GPCRs and Schild analysis. Presented at SBS 2006 (Society of Biomolecular Sciences Conference) | | | | | | The CellKey™ assay system can be used as a simple and fast functional assay for the measurement of chemokine receptor-ligand binding activity. In this poster, the CellKey™ system was used to analyze the activity of CXCR4 in several human cancer cell lines. The CXCR4 receptor is involved in many biological processes including hematopoiesis, immune cell trafficking, malignant cell growth, and metastasis. The EC 50 and IC 50 values generated were consistent with literature values. Additionally, the endogenous CXCR4 is known to be Gi-coupled, and this was confirmed with specific modulators. Presented at SBS 2006 (Society of Biomolecular Sciences Conference) | | | | | | b-adrenoreceptor antagonists are one of the most widely used classes of drugs in
the management of hypertension, heart disease, anxiety and migraine. Additionally,
b2-adrenoreceptor agonists are used in the treatment of asthma and chronic obstructive pulmonary disease. Identifying antagonists that are selective for the different b1, b2 or b3 subtypes is important in the control of deleterious side effects. In this study, the CellKey™ system is used to measure endogenous b-adrenoreceptor activity in adherent and non-adherent cells, identify the receptor subtype expressed with subtype-selective agonists, and generate pharmacologically relevant data including Schild analysis. Presented at DDT 2006 (Drug Discovery Technology Conference) | | | | | | CDS was used to perform typical hit-to-lead experiments on a panel of compounds applied to two different GPCR targets. Hit confirmation and pharmacological data were generated for agonist and antagonist compounds discovered in an HTS screening campaign. CDS response profiles were examined to gain information about the manner in which each compound interacted with the cells, including the identification of non-silent antagonists and of non-selective compounds. Presented at SBS 2005 (Society of Biomolecular Sciences Conference), IIR Cell-Based Assay 2006 Conference and Screentech 2006 | | | | | | The ability to measure the activity of endogenous receptors in a live-cell kinetic assay is attractive in drug discovery because it permits the characterization of receptors in their native environment. The ability to use human primary cells is of particular interest because they provide an in vivo model that is physiologically identical to human tissue. The CellKey™ system, monitors cellular physiology in a non-invasive manner and without the need for fluorophores, tagged proteins or transfected reporter systems. To illustrate the power of this label-free approach, we present data for a number of receptors in primary human prostate cells (epithelial, stromal and smooth muscle). Using the CellKey™ platform, activation of G-protein coupled receptors (GPCRs) results in CDS response profiles that are characteristic of their functional G-protein coupling mechanism. Proprietary cluster analysis of CDS response profiles may be used in the automated identification of the G-protein coupling of uncharacterized GPCRs. In prostate cells, pharmacological evaluations were performed to rank a panel of muscarinic agonists and determine EC50 and IC50 values for an agonist and antagonist to the EGF receptor. These studies detail powerful techniques that can be applied to selectivity screening of compounds and target validation in an endogenous setting. Presented at CHI RNAi 2006 | | | | | | CDS was used to conduct a pharmacological evaluation of the activation of an endogenously expressed GPCR in two related rhabdomyosarcoma cell lines. CDS was able to consistently measure activation of the endogenous receptor. In contrast, Ca2+ assays required the development of a clonal cell line and the use of non-physiological concentrations of agonist to elicit a response. Presented at SBS 2005 (Society of Biomolecular Sciences Conference), Screentech 2006 and IIR GPCR 2006 Conference | | | | | | This poster presents the use of CDS in the pharmacological evaluation of ligands for a number of receptors in a variety of cell types. Data on hit confirmation, receptor selectivity analysis, ligand potency, and Schild analysis of receptor-selective antagonists is presented. The study demonstrates that CDS measurements quantitatively align with results from other cell-based assays in determining the potency and ranking of agonists and antagonists. Presented at DDT 2005 (Drug Discovery Technology Conference) | |
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