The Experts below are selected from a list of 20421 Experts worldwide ranked by ideXlab platform

Craig J Thomas - One of the best experts on this subject based on the ideXlab platform.

  • a homogeneous sirpα cd47 cell based Ligand Binding Assay utility for small molecule drug development in immuno oncology
    PLOS ONE, 2020
    Co-Authors: Teresa L Burgess, Joshua D Amason, Jeffrey S Rubin, Damien Y Duveau, Laurence Lamy, David D Roberts, Catherine L Farrell, James Inglese, Craig J Thomas
    Abstract:

    CD47 is an immune checkpoint protein that downregulates both the innate and adaptive anti-tumor immune response via its counter receptor SIRPα. Biologics, including humanized CD47 monoclonal antibodies and decoy SIRPα receptors, that block the SIRPα-CD47 interaction, are currently being developed as cancer immunotherapy agents. However, adverse side effects and limited penetration of tumor tissue associated with their structure and large size may impede their clinical application. We recently developed a quantitative high throughput screening Assay platform to identify small molecules that disrupt the Binding of SIRPα and CD47 as an alternative approach to these protein-based therapeutics. Here, we report on the development and optimization of a cell-based Binding Assay to validate active small molecules from our biochemical screening effort. This Assay has a low volume, high capacity homogenous format that relies on laser scanning cytometry (LSC) and associated techniques to enhance signal to noise measurement of cell surface Binding. The LSC Assay is specific, concentration dependent, and validated for the two major human SIRPα variants (V1 and V2), with results that parallel those of our biochemical data as well as published studies. We also utilized the LSC Assay to confirm published studies showing that the inhibition of amino-terminal pyroglutamate formation on CD47 using the glutaminyl cyclase inhibitor SEN177 disrupts SIRPα Binding. The SIRPα-CD47 interaction could be quantitatively measured in live and fixed tumor cells. Use of fixed cells reduces the burden of cell maintenance and provides stable cell standards to control for inter- and intra-Assay variations. We also demonstrate the utility of the Assay to characterize the activity of the first reported small molecule antagonists of the SIRPα-CD47 interaction. This Assay will support the screening of thousands of compounds to identify or validate active small molecules as hits, develop structure activity relationships and assist in the optimization of hits to leads by a typical iterative medicinal chemistry campaign.

  • a homogeneous sirpα cd47 cell based Ligand Binding Assay utility for small molecule drug development in immuno oncology
    bioRxiv, 2019
    Co-Authors: Teresa L Burgess, Joshua D Amason, Jeffrey S Rubin, Damien Y Duveau, Laurence Lamy, David D Roberts, Catherine L Farrell, James Inglese, Craig J Thomas, Thomas W Miller
    Abstract:

    ABSTRACT CD47 is an immune checkpoint protein that downregulates both the innate and adaptive anti-tumor immune response via its counter receptor SIRPα. Biologics, including humanized CD47 monoclonal antibodies and decoy SIRPα receptors, that block the SIRPα-CD47 interaction, are currently being developed as cancer immunotherapy agents. However, adverse side effects and limited penetration of tumor tissue associated with their structure and large size may impede their clinical application. We recently developed a quantitative high throughput screening Assay platform to identify small molecules that disrupt the Binding of SIRPα and CD47 as an alternative approach to these protein-based therapeutics.  Here, we report on the development and optimization of a cell-based Binding Assay to validate active small molecules from our biochemical screening effort. This Assay has a low volume, high capacity homogenous format that relies on laser scanning cytometry (LSC) and associated techniques to enhance signal to noise measurement of cell surface Binding. The LSC Assay is specific, concentration dependent, and validated for the two major human SIRPα variants (V1 and V2), with results that parallel those of our biochemical data as well as published studies.  The LSC Assay also confirmed published studies, such as the inhibition of amino-terminal pyroglutamate formation in CD47 disrupts SIRPα Binding. SIRPα-CD47 interaction could be quantitatively measured in live and fixed tumor cells. Use of fixed cells reduces the burden of cell maintenance and provides stable cell standards to control for inter- and intra-Assay variations.   We also demonstrate the utility of the Assay to characterize the activity of the first reported small molecule antagonists of the SIRPα-CD47 interaction. This Assay will support the screening of thousands of compounds to identify or validate active small molecules as hits, develop structure activity relationships and assist in the optimization of hits to leads by a typical iterative medicinal chemistry campaign.

Joshua D Amason - One of the best experts on this subject based on the ideXlab platform.

  • a homogeneous sirpα cd47 cell based Ligand Binding Assay utility for small molecule drug development in immuno oncology
    PLOS ONE, 2020
    Co-Authors: Teresa L Burgess, Joshua D Amason, Jeffrey S Rubin, Damien Y Duveau, Laurence Lamy, David D Roberts, Catherine L Farrell, James Inglese, Craig J Thomas
    Abstract:

    CD47 is an immune checkpoint protein that downregulates both the innate and adaptive anti-tumor immune response via its counter receptor SIRPα. Biologics, including humanized CD47 monoclonal antibodies and decoy SIRPα receptors, that block the SIRPα-CD47 interaction, are currently being developed as cancer immunotherapy agents. However, adverse side effects and limited penetration of tumor tissue associated with their structure and large size may impede their clinical application. We recently developed a quantitative high throughput screening Assay platform to identify small molecules that disrupt the Binding of SIRPα and CD47 as an alternative approach to these protein-based therapeutics. Here, we report on the development and optimization of a cell-based Binding Assay to validate active small molecules from our biochemical screening effort. This Assay has a low volume, high capacity homogenous format that relies on laser scanning cytometry (LSC) and associated techniques to enhance signal to noise measurement of cell surface Binding. The LSC Assay is specific, concentration dependent, and validated for the two major human SIRPα variants (V1 and V2), with results that parallel those of our biochemical data as well as published studies. We also utilized the LSC Assay to confirm published studies showing that the inhibition of amino-terminal pyroglutamate formation on CD47 using the glutaminyl cyclase inhibitor SEN177 disrupts SIRPα Binding. The SIRPα-CD47 interaction could be quantitatively measured in live and fixed tumor cells. Use of fixed cells reduces the burden of cell maintenance and provides stable cell standards to control for inter- and intra-Assay variations. We also demonstrate the utility of the Assay to characterize the activity of the first reported small molecule antagonists of the SIRPα-CD47 interaction. This Assay will support the screening of thousands of compounds to identify or validate active small molecules as hits, develop structure activity relationships and assist in the optimization of hits to leads by a typical iterative medicinal chemistry campaign.

  • a homogeneous sirpα cd47 cell based Ligand Binding Assay utility for small molecule drug development in immuno oncology
    bioRxiv, 2019
    Co-Authors: Teresa L Burgess, Joshua D Amason, Jeffrey S Rubin, Damien Y Duveau, Laurence Lamy, David D Roberts, Catherine L Farrell, James Inglese, Craig J Thomas, Thomas W Miller
    Abstract:

    ABSTRACT CD47 is an immune checkpoint protein that downregulates both the innate and adaptive anti-tumor immune response via its counter receptor SIRPα. Biologics, including humanized CD47 monoclonal antibodies and decoy SIRPα receptors, that block the SIRPα-CD47 interaction, are currently being developed as cancer immunotherapy agents. However, adverse side effects and limited penetration of tumor tissue associated with their structure and large size may impede their clinical application. We recently developed a quantitative high throughput screening Assay platform to identify small molecules that disrupt the Binding of SIRPα and CD47 as an alternative approach to these protein-based therapeutics.  Here, we report on the development and optimization of a cell-based Binding Assay to validate active small molecules from our biochemical screening effort. This Assay has a low volume, high capacity homogenous format that relies on laser scanning cytometry (LSC) and associated techniques to enhance signal to noise measurement of cell surface Binding. The LSC Assay is specific, concentration dependent, and validated for the two major human SIRPα variants (V1 and V2), with results that parallel those of our biochemical data as well as published studies.  The LSC Assay also confirmed published studies, such as the inhibition of amino-terminal pyroglutamate formation in CD47 disrupts SIRPα Binding. SIRPα-CD47 interaction could be quantitatively measured in live and fixed tumor cells. Use of fixed cells reduces the burden of cell maintenance and provides stable cell standards to control for inter- and intra-Assay variations.   We also demonstrate the utility of the Assay to characterize the activity of the first reported small molecule antagonists of the SIRPα-CD47 interaction. This Assay will support the screening of thousands of compounds to identify or validate active small molecules as hits, develop structure activity relationships and assist in the optimization of hits to leads by a typical iterative medicinal chemistry campaign.

Teresa L Burgess - One of the best experts on this subject based on the ideXlab platform.

  • a homogeneous sirpα cd47 cell based Ligand Binding Assay utility for small molecule drug development in immuno oncology
    PLOS ONE, 2020
    Co-Authors: Teresa L Burgess, Joshua D Amason, Jeffrey S Rubin, Damien Y Duveau, Laurence Lamy, David D Roberts, Catherine L Farrell, James Inglese, Craig J Thomas
    Abstract:

    CD47 is an immune checkpoint protein that downregulates both the innate and adaptive anti-tumor immune response via its counter receptor SIRPα. Biologics, including humanized CD47 monoclonal antibodies and decoy SIRPα receptors, that block the SIRPα-CD47 interaction, are currently being developed as cancer immunotherapy agents. However, adverse side effects and limited penetration of tumor tissue associated with their structure and large size may impede their clinical application. We recently developed a quantitative high throughput screening Assay platform to identify small molecules that disrupt the Binding of SIRPα and CD47 as an alternative approach to these protein-based therapeutics. Here, we report on the development and optimization of a cell-based Binding Assay to validate active small molecules from our biochemical screening effort. This Assay has a low volume, high capacity homogenous format that relies on laser scanning cytometry (LSC) and associated techniques to enhance signal to noise measurement of cell surface Binding. The LSC Assay is specific, concentration dependent, and validated for the two major human SIRPα variants (V1 and V2), with results that parallel those of our biochemical data as well as published studies. We also utilized the LSC Assay to confirm published studies showing that the inhibition of amino-terminal pyroglutamate formation on CD47 using the glutaminyl cyclase inhibitor SEN177 disrupts SIRPα Binding. The SIRPα-CD47 interaction could be quantitatively measured in live and fixed tumor cells. Use of fixed cells reduces the burden of cell maintenance and provides stable cell standards to control for inter- and intra-Assay variations. We also demonstrate the utility of the Assay to characterize the activity of the first reported small molecule antagonists of the SIRPα-CD47 interaction. This Assay will support the screening of thousands of compounds to identify or validate active small molecules as hits, develop structure activity relationships and assist in the optimization of hits to leads by a typical iterative medicinal chemistry campaign.

  • a homogeneous sirpα cd47 cell based Ligand Binding Assay utility for small molecule drug development in immuno oncology
    bioRxiv, 2019
    Co-Authors: Teresa L Burgess, Joshua D Amason, Jeffrey S Rubin, Damien Y Duveau, Laurence Lamy, David D Roberts, Catherine L Farrell, James Inglese, Craig J Thomas, Thomas W Miller
    Abstract:

    ABSTRACT CD47 is an immune checkpoint protein that downregulates both the innate and adaptive anti-tumor immune response via its counter receptor SIRPα. Biologics, including humanized CD47 monoclonal antibodies and decoy SIRPα receptors, that block the SIRPα-CD47 interaction, are currently being developed as cancer immunotherapy agents. However, adverse side effects and limited penetration of tumor tissue associated with their structure and large size may impede their clinical application. We recently developed a quantitative high throughput screening Assay platform to identify small molecules that disrupt the Binding of SIRPα and CD47 as an alternative approach to these protein-based therapeutics.  Here, we report on the development and optimization of a cell-based Binding Assay to validate active small molecules from our biochemical screening effort. This Assay has a low volume, high capacity homogenous format that relies on laser scanning cytometry (LSC) and associated techniques to enhance signal to noise measurement of cell surface Binding. The LSC Assay is specific, concentration dependent, and validated for the two major human SIRPα variants (V1 and V2), with results that parallel those of our biochemical data as well as published studies.  The LSC Assay also confirmed published studies, such as the inhibition of amino-terminal pyroglutamate formation in CD47 disrupts SIRPα Binding. SIRPα-CD47 interaction could be quantitatively measured in live and fixed tumor cells. Use of fixed cells reduces the burden of cell maintenance and provides stable cell standards to control for inter- and intra-Assay variations.   We also demonstrate the utility of the Assay to characterize the activity of the first reported small molecule antagonists of the SIRPα-CD47 interaction. This Assay will support the screening of thousands of compounds to identify or validate active small molecules as hits, develop structure activity relationships and assist in the optimization of hits to leads by a typical iterative medicinal chemistry campaign.

Christa E. Müller - One of the best experts on this subject based on the ideXlab platform.

  • fluorescent labeled selective adenosine a2b receptor antagonist enables competition Binding Assay by flow cytometry
    Journal of Medicinal Chemistry, 2018
    Co-Authors: Meryem Kose, Sabrina Gollos, Tadeusz Karcz, Amelie Fiene, Fabian Heisig, Andrea Behrenswerth, Vigneshwaran Namasivayam, Katarzyna Kieckononowicz, Christa E. Müller
    Abstract:

    Fluorescent Ligands represent powerful tools for biological studies and are considered attractive alternatives to radioLigands. In this study, we developed fluorescent antagonists for A2B adenosine receptors (A2BARs), which are targeted by antiasthmatic xanthines and were proposed as novel targets in immuno-oncology. Our approach was to merge a small borondipyrromethene (BODIPY) derivative with the pharmacophore of 8-substituted xanthine derivatives. On the basis of the design, synthesis, and evaluation of model compounds, several fluorescent Ligands were synthesized. Compound 29 (PSB-12105), which displayed high affinity for human, rat, and mouse A2BARs (Ki = 0.2–2 nM) and high selectivity for this AR subtype, was selected for further studies. A homology model of the human A2BAR was generated, and docking studies were performed. Moreover, 29 allowed us to establish a homogeneous receptor–Ligand Binding Assay using flow cytometry. These compounds constitute the first potent, selective fluorescent A2BAR li...

Stefan Hannus - One of the best experts on this subject based on the ideXlab platform.

  • homogeneous time resolved g protein coupled receptor Ligand Binding Assay based on fluorescence cross correlation spectroscopy
    Analytical Biochemistry, 2016
    Co-Authors: Thomas Antoine, David Ott, Katharina Ebell, Kerrin Hansen, Luc Henry, Frank Becker, Stefan Hannus
    Abstract:

    G protein-coupled receptors (GPCRs) mediate many important physiological functions and are considered as one of the most successful therapeutic target classes for a wide spectrum of diseases. Drug discovery projects generally benefit from a broad range of experimental approaches for screening compound libraries and for the characterization of Binding modes of drug candidates. Owing to the difficulties in solubilizing and purifying GPCRs, Assay formats have been so far mainly limited to cell-based functional Assays and radioLigand Binding Assays. In this study, we used fluorescence cross-correlation spectroscopy (FCCS) to analyze the interaction of detergent-solubilized receptors to various types of GPCR Ligands: endogenous peptides, small molecules, and a large surrogate antagonist represented by a blocking monoclonal antibody. Our work demonstrates the suitability of the homogeneous and time-resolved FCCS Assay format for a robust, high-throughput determination of receptor-Ligand Binding affinities and kinetic rate constants for various therapeutically relevant GPCRs.