Flow Virometry to Analyze Antigenic Spectra of Virions and Extracellular Vesicles

Overview

Journal J Vis Exp

Specialties Biology
General Medicine

Date 2017 Feb 13

PMID 28190041

Citations 4

Authors

Anush Arakelyan

Wendy Fitzgerald

Sonia Zicari

Murad Vagida

Jean-Charles Grivel

Leonid Margolis

Affiliations

Soon will be listed here.

Abstract

Cells release small extracellular vesicles (EVs) into the surrounding media. Upon virus infection cells also release virions that have the same size of some of the EVs. Both virions and EVs carry proteins of the cells that generated them and are antigenically heterogeneous. In spite of their diversity, both viruses and EVs were characterized predominantly by bulk analysis. Here, we describe an original nanotechnology-based high throughput method that allows the characterization of antigens on individual small particles using regular flow cytometers. Viruses or extracellular vesicles were immunocaptured with 15 nm magnetic nanoparticles (MNPs) coupled to antibodies recognizing one of the surface antigens. The captured virions or vesicles were incubated with fluorescent antibodies against other surface antigens. The resultant complexes were separated on magnetic columns from unbound antibodies and analyzed with conventional flow cytometers triggered on fluorescence. This method has wide applications and can be used to characterize the antigenic composition of any viral- and non-viral small particles generated by cells in vivo and in vitro. Here, we provide examples of the usage of this method to evaluate the distribution of host cell markers on individual HIV-1 particles, to study the maturation of individual Dengue virions (DENV), and to investigate extracellular vesicles released into the bloodstream.

Citing Articles

Applying Flow Virometry to Study the HIV Envelope Glycoprotein and Differences Across HIV Model Systems.

Burnie J, Fernandes C, Patel A, Persaud A, Chaphekar D, Wei D Viruses. 2024; 16(6).

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Harnessing the Therapeutic Potential of Extracellular Vesicles for Biomedical Applications Using Multifunctional Magnetic Nanomaterials.

Yang L, Patel K, Rathnam C, Thangam R, Hou Y, Kang H Small. 2022; 18(13):e2104783.

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Flow virometry for process monitoring of live virus vaccines-lessons learned from ERVEBO.

Ricci G, Minsker K, Kapish A, Osborn J, Ha S, Davide J Sci Rep. 2021; 11(1):7432.

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Flow Virometry: a Powerful Tool To Functionally Characterize Viruses.

Lippe R J Virol. 2017; 92(3).

PMID: 29167334 PMC: 5774884. DOI: 10.1128/JVI.01765-17.

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