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Profiling of Surfac...
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Sahi, MaryamKTH,Affinitets-proteomik,Science for Life Laboratory, SciLifeLab
(author)
Profiling of Surface Protein Epitopes on Viral Particles by Multiplex Dual-Reporter Strategy
- Article/chapterEnglish2024
Publisher, publication year, extent ...
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MyJove Corporation,2024
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printrdacarrier
Numbers
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LIBRIS-ID:oai:DiVA.org:kth-343495
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https://urn.kb.se/resolve?urn=urn:nbn:se:kth:diva-343495URI
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https://doi.org/10.3791/66230DOI
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http://kipublications.ki.se/Default.aspx?queryparsed=id:238284526URI
Supplementary language notes
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Language:English
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Summary in:English
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Classification
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Subject category:ref swepub-contenttype
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Subject category:art swepub-publicationtype
Notes
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QC 20240215
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Membrane proteins on enveloped viruses play an important role in many biological functions involving virus attachment to target cell receptors, fusion of viral particles to host cells, host-virus interactions, and disease pathogenesis. Furthermore, viral membrane proteins on virus particles and presented on host cell surfaces have proven to be excellent targets for antivirals and vaccines. Here, we describe a protocol to investigate surface proteins on intact severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) particles using the dual-reporter flow cytometric system. The assay exploits multiplex technology to obtain a triple detection of viral particles by three independent affinity reactions. Magnetic beads conjugated to recombinant human angiotensin-converting enzyme-2 (ACE2) were used to capture viral particles from the supernatant of cells infected with SARS-CoV-2. Then, two detection reagents labeled with R-phycoerythrin (PE) or Brilliant Violet 421 (BV421) were applied simultaneously. As a proof-of-concept, antibody fragments targeting different epitopes of the SARS CoV-2 surface protein Spike (S1) were used. The detection of viral particles by three independent affinity reactions provides strong specificity and confirms the capture of intact virus particles. Dose-dependency curves of SARS-CoV-2 infected cell supernatant were generated with replicate coefficient variances (mean/SD) ˂14%. Good assay performance in both channels confirmed that two virus surface target protein epitopes are detectable in parallel. The protocol described here could be applied for (i) high-multiplex, high-throughput profiling of surface proteins expressed on enveloped viruses; ii) detection of active intact viral particles; and (iii) assessment of specificity and affinity of antibodies and antiviral drugs for surface epitopes of viral antigens.The application can be potentially extended to any type of extracellular vesicles and bioparticles, exposing surface antigens in body fluids or other liquid matrices.
Subject headings and genre
Added entries (persons, corporate bodies, meetings, titles ...)
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Andersson, SarahKTH,Affinitets-proteomik,Science for Life Laboratory, SciLifeLab(Swepub:kth)u1gmxc8k
(author)
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Mattsson, CeciliaKTH,Affinitets-proteomik,Science for Life Laboratory, SciLifeLab(Swepub:kth)u1adnwkp
(author)
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Dale, MatildaKTH,Affinitets-proteomik,Science for Life Laboratory, SciLifeLab(Swepub:kth)u1n0viuj
(author)
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Kagiolglou, SofiaKTH,Affinitets-proteomik,Science for Life Laboratory, SciLifeLab(Swepub:kth)PI000000
(author)
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Hofström, CamillaKTH,Science for Life Laboratory, SciLifeLab,Läkemedelsutveckling(Swepub:kth)u12237py
(author)
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Persson, HelenaKTH,Science for Life Laboratory, SciLifeLab,Läkemedelsutveckling(Swepub:kth)u1l700sh
(author)
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Klingström, JonasCenter for Infectious Medicine, Department of Medicine Huddinge, Karolinska Institutet, Sweden; Public Health Agency of Sweden, Sweden
(author)
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Chiodi, FrancescaKarolinska Institutet
(author)
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Fredolini, ClaudiaKTH,Science for Life Laboratory, SciLifeLab,Affinitets-proteomik(Swepub:kth)u11xul3s
(author)
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KTHAffinitets-proteomik
(creator_code:org_t)
Related titles
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In:Journal of Visualized Experiments: MyJove Corporation2024:2031940-087X
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Sahi, Maryam
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Andersson, Sarah
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Mattsson, Cecili ...
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Dale, Matilda
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Kagiolglou, Sofi ...
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Hofström, Camill ...
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Persson, Helena
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Klingström, Jona ...
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Chiodi, Francesc ...
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Fredolini, Claud ...
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- About the subject
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- NATURAL SCIENCES
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NATURAL SCIENCES
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and Biological Scien ...
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and Biochemistry and ...
- Articles in the publication
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Journal of Visua ...
- By the university
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Royal Institute of Technology
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Karolinska Institutet