Microarray Compound Screening (μARCS) to identify inhibitors of HIV integrase

Caroline A. David, Tim Middleton, Debra Montgomery, Hock Ben Lim, Warren Kati, Akhter Molla, Xiaoling Xuei, Usha Warrior, James L. Kofron, David J. Burns

Research output: Contribution to journalArticle

27 Citations (Scopus)

Abstract

A novel high-throughput strand transfer assay has been developed, using Microarray Compound Screening (μARCS) technology, to identify inhibitors of human immunodeficiency virus (HIV) integrase. This technology utilizes agarose matrices to introduce a majority of the reagents throughout the assay. Integration of biotinylated donor DNA with fluorescein isothiocyanate (FITC)-labeled target DNA occurs on a SAM membrane in the presence of integrase. An anti-FITC antibody conjugated to alkaline phosphatase (AP) was used to do an enzyme-linked immunosorbent assay with the SAM. An agarose gel containing AttoPhos, a substrate of AP, was used for detection of the integrase reactions on the SAM. For detection, the AttoPhos gel was separated from the SAM after incubation and then the gel was imaged using an Eagle Eye II closed-circuit device camera system. Potential integrase inhibitors appear as dark spots on the gel image. A library of approximately 250,000 compounds was screened using this HIV integrase strand transfer assay in μARCS format. Compounds from different structural classes were identified in this assay as novel integrase inhibitors.

Original languageEnglish (US)
Pages (from-to)259-266
Number of pages8
JournalJournal of Biomolecular Screening
Volume7
Issue number3
DOIs
StatePublished - 2002
Externally publishedYes

Fingerprint

Integrases
Microarrays
Viruses
Assays
Screening
Gels
Integrase Inhibitors
HIV
Fluorescein
Sepharose
Alkaline Phosphatase
Technology
DNA
Immunosorbents
Enzyme-Linked Immunosorbent Assay
Equipment and Supplies
Cameras
Membranes
Throughput
Arthrogryposis renal dysfunction cholestasis syndrome

ASJC Scopus subject areas

  • Analytical Chemistry
  • Clinical Biochemistry
  • Biotechnology
  • Biochemistry
  • Molecular Biology

Cite this

David, C. A., Middleton, T., Montgomery, D., Lim, H. B., Kati, W., Molla, A., ... Burns, D. J. (2002). Microarray Compound Screening (μARCS) to identify inhibitors of HIV integrase. Journal of Biomolecular Screening, 7(3), 259-266. https://doi.org/10.1089/108705702760047754

Microarray Compound Screening (μARCS) to identify inhibitors of HIV integrase. / David, Caroline A.; Middleton, Tim; Montgomery, Debra; Lim, Hock Ben; Kati, Warren; Molla, Akhter; Xuei, Xiaoling; Warrior, Usha; Kofron, James L.; Burns, David J.

In: Journal of Biomolecular Screening, Vol. 7, No. 3, 2002, p. 259-266.

Research output: Contribution to journalArticle

David, CA, Middleton, T, Montgomery, D, Lim, HB, Kati, W, Molla, A, Xuei, X, Warrior, U, Kofron, JL & Burns, DJ 2002, 'Microarray Compound Screening (μARCS) to identify inhibitors of HIV integrase', Journal of Biomolecular Screening, vol. 7, no. 3, pp. 259-266. https://doi.org/10.1089/108705702760047754
David, Caroline A. ; Middleton, Tim ; Montgomery, Debra ; Lim, Hock Ben ; Kati, Warren ; Molla, Akhter ; Xuei, Xiaoling ; Warrior, Usha ; Kofron, James L. ; Burns, David J. / Microarray Compound Screening (μARCS) to identify inhibitors of HIV integrase. In: Journal of Biomolecular Screening. 2002 ; Vol. 7, No. 3. pp. 259-266.
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