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Nanoparticle-based flow virometry for the analysis of individual virions
Anush Arakelyan, … , Leonid Margolis, Jean-Charles Grivel
Anush Arakelyan, … , Leonid Margolis, Jean-Charles Grivel
Published August 8, 2013
Citation Information: J Clin Invest. 2013;123(9):3716-3727. https://doi.org/10.1172/JCI67042.
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Technical Advance AIDS/HIV Article has an altmetric score of 19

Nanoparticle-based flow virometry for the analysis of individual virions

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Abstract

While flow cytometry has been used to analyze the antigenic composition of individual cells, the antigenic makeup of viral particles is still characterized predominantly in bulk. Here, we describe a technology, “flow virometry,” that can be used for antigen detection on individual virions. The technology is based on binding magnetic nanoparticles to virions, staining the virions with monoclonal antibodies, separating the formed complexes with magnetic columns, and characterizing them with flow cytometers. We used this technology to study the distribution of two antigens (HLA-DR and LFA-1) that HIV-1 acquires from infected cells among individual HIV-1 virions. Flow virometry revealed that the antigenic makeup of virions from a single preparation is heterogeneous. This heterogeneity could not be detected with bulk analysis of viruses. Moreover, in two preparations of the same HIV-1 produced by different cells, the distribution of antigens among virions was different. In contrast, HIV-1 of two different HIV-1 genotypes replicating in the same cells became somewhat antigenically similar. This nanotechnology allows the study of virions in bodily fluids without virus propagation and in principle is not restricted to the analysis of HIV, but can be applied to the analysis of the individual surface antigenic makeup of any virus.

Authors

Anush Arakelyan, Wendy Fitzgerald, Leonid Margolis, Jean-Charles Grivel

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Figure 6

Distribution of cellular antigens on HIV-1LAI.04 and HIV-1SF162 produced in two different cell types.

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Distribution of cellular antigens on HIV-1LAI.04 and HIV-1SF162 produced...
HIV-1LAI.04 and HIV-1SF162 were produced either in PBMCs or in Jurkat cells. Preparations of HIV-1 were captured on MNPs via VRC01, and virions were visualized with Alexa Fluor 647–labeled 2G12. The presence of HLA-DR (white bars) and LFA-1 (black bars) were revealed with PE-Cy5.5– and PE-Cy7–labeled antibodies specific to these antigens, respectively. Presented are the averages ± SEM of three to six experiments.

Copyright © 2025 American Society for Clinical Investigation
ISSN: 0021-9738 (print), 1558-8238 (online)

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