A novel high throughput quantum dot-based fluorescence assay for quantitation of virus binding and attachment

Quantum dots (QDots) are fluorescent semiconductor nanocrystals with a narrow emission spectrum, high quantum yield, and excellent photostability. These unique properties of QDots have been utilized to develop a fluorescent binding assay using biotinylated human T cell leukemia virus type 1 (biot-HT...

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Bibliographic Details
Published in:Journal of virological methods 2007-05, Vol.141 (2), p.125-132
Main Authors: Kampani, Karan, Quann, Kevin, Ahuja, Jaya, Wigdahl, Brian, Khan, Zafar K., Jain, Pooja
Format: Article
Language:English
Subjects:
Biological and medical sciences
Biotinylation
Cell Line
Cells, Cultured
Fluorescence
Fundamental and applied biological sciences. Psychology
HTLV-1
Human T-lymphotropic virus 1
Human T-lymphotropic virus 1 - physiology
Humans
Leukocytes, Mononuclear
Microbiology
Microscopy, Confocal - methods
Quantum dot
Quantum Dots
Techniques used in virology
Viral binding assay
Virology
Virus Attachment
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Summary:Quantum dots (QDots) are fluorescent semiconductor nanocrystals with a narrow emission spectrum, high quantum yield, and excellent photostability. These unique properties of QDots have been utilized to develop a fluorescent binding assay using biotinylated human T cell leukemia virus type 1 (biot-HTLV-1) conjugated with streptavidin-coated QDots that enabled both qualitative and quantitative analyses of viral binding. The specificity and linearity of the assay was demonstrated utilizing T cells, the primary HTLV-1-susceptible cell population. Furthermore, differential binding of HTLV-1 was analyzed in additional cell types of clinical relevance including primary CD4 + and CD8 + T cells, dendritic cells (DCs), monocytes, bone marrow progenitor cells, and epithelial cells. DCs exhibited maximum binding affinity when compared to other examined cell types except the Jurkat and SUP-T1 T cell lines. Finally, blocking antibodies directed against a putative HTLV-1 receptor on DCs; DC-SIGN (dendritic cell-specific ICAM-3-grabbing non-integrin), were utilized to study the inhibition of HTLV-1 binding to target cells. Overall, these results demonstrated that this novel high throughput assay can be utilized to study the binding of a biotinylated virus and has implications for screening of viral binding inhibitors as well as host membrane proteins that may serve as receptors for viral entry.
ISSN:0166-0934
1879-0984
DOI:10.1016/j.jviromet.2006.11.043