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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

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 3

Single particle detection.

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Single particle detection.
(A) Two preparations of labeled MNPs, one com...
(A) Two preparations of labeled MNPs, one complexed with PE-Cy7–labeled antibody (left panel) and the other with a PE-Cy5.5–labeled antibody (center panel) were mixed (right panel). Of the events recorded, approximately 1.5% appeared to be double positive (right panel). Thus, approximately 98.5% of events represent single MNPs. Results represent one of four experiments. (B) A preparation of HIV-1SF162 was divided into two parts: one part was stained with DiD (left panel) and the other with DiO (center panel). The two preparations were mixed, and the virions were captured by MNPs and analyzed (right panel). Note, about 90% of the events represent individual HIV-1 virions. Results represent one of four experiments. On each plot, the fraction of events in their respective gates is expressed as a percentage of the total events in the plot.

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ISSN: 0021-9738 (print), 1558-8238 (online)

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