Skip to main page content
U.S. flag

An official website of the United States government

Dot gov

The .gov means it’s official.
Federal government websites often end in .gov or .mil. Before sharing sensitive information, make sure you’re on a federal government site.

Https

The site is secure.
The https:// ensures that you are connecting to the official website and that any information you provide is encrypted and transmitted securely.

Access keys NCBI Homepage MyNCBI Homepage Main Content Main Navigation
. 2005 Jun;10(4):374-82.
doi: 10.1177/1087057105274532.

High-throughput screening with HyperCyt flow cytometry to detect small molecule formylpeptide receptor ligands

Susan M Young  1 , Cristian Bologa, Eric R Prossnitz, Tudor I Oprea, Larry A Sklar, Bruce S Edwards
Affiliations

High-throughput screening with HyperCyt flow cytometry to detect small molecule formylpeptide receptor ligands

Susan M Young et al. J Biomol Screen. 2005 Jun.

Abstract

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 (approximately 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 [K(i)]=14 microM) 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.

PubMed Disclaimer

Similar articles

See all similar articles

Cited by

  • Optimization of Peptide Linker-Based Fluorescent Ligands for the Histamine H1 Receptor.
    Kok ZY, Stoddart LA, Mistry SJ, Mocking TAM, Vischer HF, Leurs R, Hill SJ, Mistry SN, Kellam B. Kok ZY, et al. J Med Chem. 2022 Jun 23;65(12):8258-8288. doi: 10.1021/acs.jmedchem.2c00125. Epub 2022 Jun 3. J Med Chem. 2022. PMID: 35734860 Free PMC article.
  • Finding New Molecular Targets of Familiar Natural Products Using In Silico Target Prediction.
    Mayr F, Möller G, Garscha U, Fischer J, Rodríguez Castaño P, Inderbinen SG, Temml V, Waltenberger B, Schwaiger S, Hartmann RW, Gege C, Martens S, Odermatt A, Pandey AV, Werz O, Adamski J, Stuppner H, Schuster D. Mayr F, et al. Int J Mol Sci. 2020 Sep 26;21(19):7102. doi: 10.3390/ijms21197102. Int J Mol Sci. 2020. PMID: 32993084 Free PMC article.
  • Fluorescent ligands: Bringing light to emerging GPCR paradigms.
    Soave M, Briddon SJ, Hill SJ, Stoddart LA. Soave M, et al. Br J Pharmacol. 2020 Mar;177(5):978-991. doi: 10.1111/bph.14953. Epub 2020 Feb 6. Br J Pharmacol. 2020. PMID: 31877233 Free PMC article. Review.
  • Guidelines for the use of flow cytometry and cell sorting in immunological studies (second edition).
    Cossarizza A, Chang HD, Radbruch A, Acs A, Adam D, Adam-Klages S, Agace WW, Aghaeepour N, Akdis M, Allez M, Almeida LN, Alvisi G, Anderson G, Andrä I, Annunziato F, Anselmo A, Bacher P, Baldari CT, Bari S, Barnaba V, Barros-Martins J, Battistini L, Bauer W, Baumgart S, Baumgarth N, Baumjohann D, Baying B, Bebawy M, Becher B, Beisker W, Benes V, Beyaert R, Blanco A, Boardman DA, Bogdan C, Borger JG, Borsellino G, Boulais PE, Bradford JA, Brenner D, Brinkman RR, Brooks AES, Busch DH, Büscher M, Bushnell TP, Calzetti F, Cameron G, Cammarata I, Cao X, Cardell SL, Casola S, Cassatella MA, Cavani A, Celada A, Chatenoud L, Chattopadhyay PK, Chow S, Christakou E, Čičin-Šain L, Clerici M, Colombo FS, Cook L, Cooke A, Cooper AM, Corbett AJ, Cosma A, Cosmi L, Coulie PG, Cumano A, Cvetkovic L, Dang VD, Dang-Heine C, Davey MS, Davies D, De Biasi S, Del Zotto G, Dela Cruz GV, Delacher M, Della Bella S, Dellabona P, Deniz G, Dessing M, Di Santo JP, Diefenbach A, Dieli F, Dolf A, Dörner T, Dress RJ, Dudziak D, Dustin M, Dutertre CA, Ebner F, Eckle SBG, Edinger M, Eede P, Ehrhardt GRA, Eich M, Engel P, Engelhardt B, Erdei A, Esser C, Everts B, Evrard M, Falk CS, Fehniger TA, Felipo-Benavent M, Fer… See abstract for full author list ➔ Cossarizza A, et al. Eur J Immunol. 2019 Oct;49(10):1457-1973. doi: 10.1002/eji.201970107. Eur J Immunol. 2019. PMID: 31633216 Free PMC article.
  • A Non-imaging High Throughput Approach to Chemical Library Screening at the Unmodified Adenosine-A3 Receptor in Living Cells.
    Arruda MA, Stoddart LA, Gherbi K, Briddon SJ, Kellam B, Hill SJ. Arruda MA, et al. Front Pharmacol. 2017 Dec 13;8:908. doi: 10.3389/fphar.2017.00908. eCollection 2017. Front Pharmacol. 2017. PMID: 29321740 Free PMC article.
See all "Cited by" articles

Publication types

LinkOut - more resources

-