Integration of Virtual Screening with High-Throughput Flow Cytometry to Identify Novel Small Molecule Formylpeptide Receptor Antagonists

2005 ◽  
Vol 68 (5) ◽  
pp. 1301-1310 ◽  
Author(s):  
Bruce S. Edwards ◽  
Cristian Bologa ◽  
Susan M. Young ◽  
Konstantin V. Balakin ◽  
Eric R. Prossnitz ◽  
...  
Keyword(s):  
Flow Cytometry ◽  
Virtual Screening ◽  
Small Molecule ◽  
High Throughput ◽  
Receptor Antagonists ◽  
Formylpeptide Receptor

Identification of Small-Molecule Inducers of FOXP3 in Human T Cells Using High-Throughput Flow Cytometry

2017 ◽  
pp. 243-252 ◽  
Author(s):  
Rob Jepras ◽  
Poonam Shah ◽  
Metul Patel ◽  
Steve Ludbrook ◽  
Gregory Wands ◽  
...  
Keyword(s):  
Flow Cytometry ◽  
T Cells ◽  
Small Molecule ◽  
High Throughput ◽  
Human T Cells

Biomolecular screening of formylpeptide receptor ligands with a sensitive, quantitative, high-throughput flow cytometry platform

2006 ◽  
Vol 1 (1) ◽  
pp. 59-66 ◽  
Author(s):  
Bruce S Edwards ◽  
Susan M Young ◽  
Tudor I Oprea ◽  
Cristian G Bologa ◽  
Eric R Prossnitz ◽  
...  
Keyword(s):  
Flow Cytometry ◽  
High Throughput ◽  
Receptor Ligands ◽  
Formylpeptide Receptor

High-Throughput Screening with HyperCyt® Flow Cytometry to Detect Small Molecule Formylpeptide Receptor Ligands

2005 ◽  
Vol 10 (4) ◽  
pp. 374-382 ◽  
Author(s):  
Susan M. Young ◽  
Cristian Bologa ◽  
Eric R. Prossnitz ◽  
Tudor I. Oprea ◽  
Larry A. Sklar ◽  
...  
Keyword(s):  
Flow Cytometry ◽  
High Throughput ◽  
High Throughput Screening ◽  
Receptor Ligands ◽  
Chemical Library ◽  
Assay Sensitivity ◽  
Compound Libraries ◽  
Analysis System ◽  
G Protein Coupled ◽  
Formylpeptide Receptor

High-throughput flow cytometry (HTFC), enabled by faster automated sample processing, represents a promising high- content approach for compound library screening. HyperCyt® is a recently developed automated HTFC analysis system by which cell samples are rapidly aspirated from microplate wells and delivered to the flow cytometer. The formylpeptide receptor (FPR) family of G protein–coupled receptors contributes to the localization and activation of tissue-damaging leukocytes at sites of chronic inflammation. Here, the authors describe development and application of an HTFC screening approach to detect potential anti-inflammatory compounds that block ligand binding to FPR. Using a homogeneous no-wash assay, samples were routinely processed at 1.5 s/well (~2500 cells analyzed/sample), allowing a 96-well plate to be processed in less than 2.5 min. Assay sensitivity and accuracy were validated by detection of a previously documented active compound with relatively low FPR affinity (sulfinpyrazone, inhibition constant [Ki]=14 μM) from among a collection of 880 compounds in the Prestwick Chemical Library. The HyperCyt® system was therefore demonstrated to be a robust, sensitive, and highly quantitative method with which to screen lead compound libraries in a 96-well format.

scholarly journals Identification of a Small GTPase Inhibitor Using a High-Throughput Flow Cytometry Bead-Based Multiplex Assay

2009 ◽  
Vol 15 (1) ◽  
pp. 10-20 ◽  
Author(s):  
Zurab Surviladze ◽  
Anna Waller ◽  
Yang Wu ◽  
Elsa Romero ◽  
Bruce S. Edwards ◽  
...  
Keyword(s):  
Flow Cytometry ◽  
Small Molecule ◽  
High Throughput ◽  
False Negative ◽  
Small Gtpases ◽  
Multiplex Assay ◽  
Small Gtpase ◽  
Dependent Manner ◽  
Gtp Binding

Small GTPases are key regulators of cellular activity and represent novel targets for the treatment of human diseases using small-molecule inhibitors. The authors describe a multiplex, flow cytometry bead-based assay for the identification and characterization of inhibitors or activators of small GTPases. Six different glutathione-S-transferase (GST)—tagged small GTPases were bound to glutathione beads, each labeled with a different red fluorescence intensity. Subsequently, beads bearing different GTPase were mixed and dispensed into 384-well plates with test compounds, and fluorescent—guanosine triphosphate (GTP) binding was used as the readout. This novel multiplex assay allowed the authors to screen a library of almost 200,000 compounds and identify more than 1200 positive compounds, which were further verified by dose-response analyses, using 6- to 8-plex assays. After the elimination of false-positive and false-negative compounds, several small-molecule families with opposing effects on GTP binding activity were identified. The authors detail the characterization of MLS000532223, a general inhibitor that prevents GTP binding to several GTPases in a dose-dependent manner and is active in biochemical and cell-based secondary assays. Live-cell imaging and confocal microscopy studies revealed the inhibitor-induced actin reorganization and cell morphology changes, characteristic of Rho GTPases inhibition. Thus, high-throughput screening via flow cytometry provides a strategy for identifying novel compounds that are active against small GTPases.

scholarly journals High‐Throughput Measurement of Small‐Molecule Enantiopurity by Using Flow Cytometry

ChemBioChem ◽  
2018 ◽  
Vol 19 (17) ◽  
pp. 1853-1857 ◽  
Author(s):  
Zhesen Tan ◽  
Jennifer M. Heemstra
Keyword(s):  
Flow Cytometry ◽  
Small Molecule ◽  
High Throughput

scholarly journals Duplex high-throughput flow cytometry screen identifies two novel formylpeptide receptor family probes

2009 ◽  
Vol 75A (3) ◽  
pp. 253-263 ◽  
Author(s):  
Susan M. Young ◽  
Cristian M. Bologa ◽  
Dan Fara ◽  
Bj K. Bryant ◽  
Juan Jacob Strouse ◽  
...  
Keyword(s):  
Flow Cytometry ◽  
High Throughput ◽  
Receptor Family ◽  
Formylpeptide Receptor
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