Volume 2 Supplement 1

2005 International Meeting of The Institute of Human Virology

Open Access

A High Throughput Quantum Dot-based Fluorescence Assay for Quantitation of HTLV-1 Binding and Attachment

  • Karan Kampani1,
  • Pooja Jain1 and
  • Brian Wigdahl1
Retrovirology20052(Suppl 1):P46

DOI: 10.1186/1742-4690-2-S1-P46

Published: 8 December 2005

Quantum dots (Qdots) are fluorescent semiconductor nanocrystals comprised of CdSe core with a semiconductor shell of zinc sulfide coated with a polymer shell allowing particles to be conjugated to biological molecules while retaining the optical properties of the particle. We have used this unique property of Qdots to develop a high throughput binding assay to study the attachment of HTLV-1 to host cells. To this end, we have biotinylated cell-free HTLV-1 (biot-HTLV-1) to facilitate viral detection using streptavidin-coated Qdots. B cells (BTHP-1 and Ramos) were exposed to biot-HTLV-1 with increasing concentrations of DEAE-dextran, a reagent known to enhance binding of other retroviruses. Unbound virus was removed by washing and cells were added with strep-Qdots and fluorescence readings were obtained at 605 nm. HTLV-1 bound efficiently to BTHP-1 and Ramos cells and this binding was significantly increased (3-fold) by DEAE-dextran. To confirm the specificity of viral binding, a competitive inhibition assay was performed wherein increasing amounts of non-biotinylated HTLV-1 was added to the binding assay along with a fixed amount of biot-HTLV-1. A dose-dependent inhibition in biot-HTLV-1 binding was observed in the presence of native virus. These results suggest that the Qdot-based assay may be useful in studying virus attachement to host cells, and the screening of inhibitors for viral binding and entry.

Notes

Authors’ Affiliations

(1)
Department of Microbiology and Immunology, and Institute for Molecular Medicine and Infectious Disease, Drexel University College of Medicine

Copyright

© The Author(s) 2005

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