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Sökning: onr:"swepub:oai:DiVA.org:kth-343495" > Profiling of Surfac...

LIBRIS Formathandbok  (Information om MARC21)
FältnamnIndikatorerMetadata
00004700naa a2200445 4500
001oai:DiVA.org:kth-343495
003SwePub
008240215s2024 | |||||||||||000 ||eng|
009oai:prod.swepub.kib.ki.se:238284526
0247 a https://urn.kb.se/resolve?urn=urn:nbn:se:kth:diva-3434952 URI
0247 a https://doi.org/10.3791/662302 DOI
0247 a http://kipublications.ki.se/Default.aspx?queryparsed=id:2382845262 URI
040 a (SwePub)kthd (SwePub)ki
041 a engb eng
042 9 SwePub
072 7a ref2 swepub-contenttype
072 7a art2 swepub-publicationtype
1001 a Sahi, Maryamu KTH,Affinitets-proteomik,Science for Life Laboratory, SciLifeLab4 aut0 (Swepub:kth)u1j5kiqj
2451 0a Profiling of Surface Protein Epitopes on Viral Particles by Multiplex Dual-Reporter Strategy
264 1b MyJove Corporation,c 2024
338 a print2 rdacarrier
500 a QC 20240215
520 a 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.
650 7a NATURVETENSKAPx Biologix Biokemi och molekylärbiologi0 (SwePub)106022 hsv//swe
650 7a NATURAL SCIENCESx Biological Sciencesx Biochemistry and Molecular Biology0 (SwePub)106022 hsv//eng
7001 a Andersson, Sarahu KTH,Affinitets-proteomik,Science for Life Laboratory, SciLifeLab4 aut0 (Swepub:kth)u1gmxc8k
7001 a Mattsson, Ceciliau KTH,Affinitets-proteomik,Science for Life Laboratory, SciLifeLab4 aut0 (Swepub:kth)u1adnwkp
7001 a Dale, Matildau KTH,Affinitets-proteomik,Science for Life Laboratory, SciLifeLab4 aut0 (Swepub:kth)u1n0viuj
7001 a Kagiolglou, Sofiau KTH,Affinitets-proteomik,Science for Life Laboratory, SciLifeLab4 aut0 (Swepub:kth)PI000000
7001 a Hofström, Camillau KTH,Science for Life Laboratory, SciLifeLab,Läkemedelsutveckling4 aut0 (Swepub:kth)u12237py
7001 a Persson, Helenau KTH,Science for Life Laboratory, SciLifeLab,Läkemedelsutveckling4 aut0 (Swepub:kth)u1l700sh
7001 a Klingström, Jonasu Center for Infectious Medicine, Department of Medicine Huddinge, Karolinska Institutet, Sweden; Public Health Agency of Sweden, Sweden4 aut
7001 a Chiodi, Francescau Karolinska Institutet4 aut
7001 a Fredolini, Claudiau KTH,Science for Life Laboratory, SciLifeLab,Affinitets-proteomik4 aut0 (Swepub:kth)u11xul3s
7102 a KTHb Affinitets-proteomik4 org
7730 t Journal of Visualized Experimentsd : MyJove Corporationg 2024:203q 2024:203x 1940-087X
8564 u https://doi.org/10.3791/66230y Fulltext
8564 8u https://urn.kb.se/resolve?urn=urn:nbn:se:kth:diva-343495
8564 8u https://doi.org/10.3791/66230
8564 8u http://kipublications.ki.se/Default.aspx?queryparsed=id:238284526

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