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Nanoparticle-based flow virometry for the analysis of individual virions
Anush Arakelyan, Wendy Fitzgerald, Leonid Margolis, Jean-Charles Grivel
Anush Arakelyan, Wendy Fitzgerald, Leonid Margolis, Jean-Charles Grivel
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Technical Advance AIDS/HIV

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 1

Outline of the flow virometry procedure.

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Outline of the flow virometry procedure.
(i) MNPs coupled to a virion-sp...
(i) MNPs coupled to a virion-specific antibody against gp120 (capture antibody, light gray). (ii) MNPs coupled to capture antibody are incubated with viruses (schematically presented as icosahedrons), which become immobilized on MNPs. (iii) MNP-immobilized viruses are visualized with human anti-gp120 antibody (blue) recognizing an epitope different from the one recognized by the capture antibody. MNPs are visualized with a fluorescent antibody or its Fab (red) against the Fc portion of the capture antibody, and viral antigens of interest are visualized with fluorescently labeled monoclonal antibodies (green and yellow). (iv) Virus-MNP complexes with bound antibodies are separated from unbound antibodies with magnetic columns. (v) The MNP-immobilized virions stained with fluorescence-labeled antibodies eluted from the magnetic columns are analyzed with the flow cytometer set up to trigger on MNP fluorescence rather than on light scatter.

Copyright © 2026 American Society for Clinical Investigation
ISSN: 0021-9738 (print), 1558-8238 (online)

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