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<p>Pseudotype virus titrations and protein expression of ACE2s and sarbecovirus Spikes.</p>
Nazia Thakur (9904162)
Joseph Newman (5638340)
Dongchun Ni (15188356)
Jeffrey Seow (9415776)
Abigail L. Hay (25157576)
Yeonjae Lee (12256119)
Ayush Upadhyay (25157573)
Babatunde E. Ekundayo (25157578)
John A. Hammond (7431785)
Thomas P. Peacock (6536120)
Kelvin Lau (1847764)
Katie J. Doores (346210)
Dalan Bailey (193037)
Microbiology
Cell Biology
Evolutionary Biology
Environmental Sciences not elsewhere classified
Ecology
Immunology
Biological Sciences not elsewhere classified
Cancer
Infectious Diseases
Virology
multiple bat coronaviruses
g ., rc
g ., civet
clade 1 viruses
restricted ace2 usage
mammalian ace2 proteins
g ., rhgb07
using monoclonal antibodies
clade 1 sarbecoviruses
div >< p
determined host range
converting enzyme 2
2 convalescent individuals
ace2 ), via
using binding assays
antigenicity using sera
related bat sarbecoviruses
using sarbecoviruses
host range
clade 3
pangolin ),
neutralising antibodies
entry assays
binding domain
sarbecoviruses interact
sarbecoviruses exhibited
rodent ace2
like rhgb07
generalist ace2
related antigenicity
zoonotic potential
vitally important
still evidence
specialist </
shifting pattern
robust evidence
research indicates
racoon dog
novel structure
largely maintained
including human
immunodominant target
human primate
generalist phenotypes
generalist </
especially omicron
entry phenotypes
diverse panel
continued evolution
breakthrough infections
<p><b>(A)</b> Lentiviral pseudotypes expressing different bat sarbecovirus Spikes were titrated on BHK-21 cells overexpressing human ACE2 (SARS-CoV-2, BANAL-20–52, BANAL-20–103, BANAL-20–236, RaTG13, SARS-CoV-1, WIV-1, Rs4231, Rf1, Rp3, RacCS203, RpYN06), <i>Rh. cornutus</i> ACE2 (Rc-o319) or <i>Rh. alcyone</i> ACE2 (RhGB07, RfGB02). Virus titres were used to normalise input virus for subsequent assays. <b>(B)</b> Expression of tagged ACE2 proteins was carried out in lysates from BHK-21 cells transfected with ACE2 constructs and probed for HA expression by Western blotting. <b>(C)</b> Sarbecovirus Spikes were used to generate lentivral-based pseudotypes, and the viral supernatant purified by ultracentrifugation under a 20% sucrose cushion. Purified viruses were probed with an anti-FLAG antibody (except RhGB07 and RfGB02, which are untagged), an anti-Spike antibody (S, full-length Spike; S1/S2, cleaved variant) and the lentivirus internal control, anti-p24 antibody. The data underlying this Figure can be found in <a href="http://www.plosbiology.org/article/info:doi/10.1371/journal.pbio.3003944#pbio.3003944.s017" target="_blank">S1 Data</a> and <a href="http://www.plosbiology.org/article/info:doi/10.1371/journal.pbio.3003944#pbio.3003944.s018" target="_blank">S1 Raw Images</a>.</p> <p>(TIF)</p>
2026-10-01T17:40:07Z
Image
Figure
10.1371/journal.pbio.3003944.s002
https://figshare.com/articles/figure/_p_Pseudotype_virus_titrations_and_protein_expression_of_ACE2s_and_sarbecovirus_Spikes_p_/34047316
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