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An in vivo NanoBiT-based pseudoviral biosensor for real-time monitoring of SARS-CoV-2 spike/hACE2 interactions and therapeutic screening.

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Title: An in vivo NanoBiT-based pseudoviral biosensor for real-time monitoring of SARS-CoV-2 spike/hACE2 interactions and therapeutic screening.
Authors: Lin, Cheng-Han1 (AUTHOR), Lin, Meng-Wei1 (AUTHOR), Yang, Xin-Rui1 (AUTHOR), Chiang, Hua-Hsin1 (AUTHOR), Wu, Ting-Hsuan1 (AUTHOR), Tsai, Chin-Hung2 (AUTHOR), Lin, Chih-Sheng1,3 (AUTHOR) lincs@nycu.edu.tw
Source: Analytica Chimica Acta. Nov2025, Vol. 1376, pN.PAG-N.PAG. 1p.
Subjects: SARS-CoV-2, Biosensors, Membrane glycoproteins, Angiotensin converting enzyme, Clinical trials, Bioluminescence, Real-time computing
Abstract: Severe acute respiratory syndrome coronavirus 2 infects host cells through its trimeric surface spike protein, which binds to the human angiotensin-converting enzyme 2 (hACE2) receptor. The spike/hACE2 interaction is a critical target for therapeutic intervention against COVID-19. Although existing hACE2 transgenic mouse (TG) models facilitate studies of viral pathogenesis, they lack the capacity for real-time, non-invasive monitoring of viral entry events in vivo. To address this limitation, we developed a novel in vivo NanoLuc Binary Technology-based pseudoviral biosensor optimized for dynamic visualization of spike–hACE2 binding in live animals. Genetically engineered LgBiT-fused hACE2 (LgBiT-hACE2) mice and a pseudovirus tagged with SmBiT-fused spike protein were developed. Upon spike–hACE2 binding, Nanoluciferase reconstitution generated quantifiable bioluminescent signals detectable within 6 h post-infection. Quantitative bioluminescence imaging using an in vivo imaging system enabled the visualization of virus infection in the LgBiT-hACE2 mouse model. Furthermore, baicalein and baicalin, two active flavonoid compounds derived from Scutellaria baicalensis , were identified as potent inhibitors for the viral spike and the hACE2 protein, respectively. Combined treatment yielded up to 90 % inhibition of spike/hACE2 interaction in vivo. This study presents an innovative in vivo biosensing model to non-invasively quantify virus–host interactions and evaluate the efficacy of spike-targeted antiviral agents under physiologically relevant conditions. [Display omitted] • In vivo NanoBiT pseudoviral biosensor monitored spike–hACE2 binding in real time. • LgBiT-hACE2 mouse model enabled non-invasive tracking of viral entry. • BA.2-SmBiT spike pseudovirus simulated SARS-CoV-2 spike-mediated infection. • NanoBiT bioluminescence imaging enhanced longitudinal infection tracking. • Baicalein and baicalin significantly inhibited spike/hACE2 interaction. [ABSTRACT FROM AUTHOR]
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Database: Engineering Source
Description
Abstract:Severe acute respiratory syndrome coronavirus 2 infects host cells through its trimeric surface spike protein, which binds to the human angiotensin-converting enzyme 2 (hACE2) receptor. The spike/hACE2 interaction is a critical target for therapeutic intervention against COVID-19. Although existing hACE2 transgenic mouse (TG) models facilitate studies of viral pathogenesis, they lack the capacity for real-time, non-invasive monitoring of viral entry events in vivo. To address this limitation, we developed a novel in vivo NanoLuc Binary Technology-based pseudoviral biosensor optimized for dynamic visualization of spike–hACE2 binding in live animals. Genetically engineered LgBiT-fused hACE2 (LgBiT-hACE2) mice and a pseudovirus tagged with SmBiT-fused spike protein were developed. Upon spike–hACE2 binding, Nanoluciferase reconstitution generated quantifiable bioluminescent signals detectable within 6 h post-infection. Quantitative bioluminescence imaging using an in vivo imaging system enabled the visualization of virus infection in the LgBiT-hACE2 mouse model. Furthermore, baicalein and baicalin, two active flavonoid compounds derived from Scutellaria baicalensis , were identified as potent inhibitors for the viral spike and the hACE2 protein, respectively. Combined treatment yielded up to 90 % inhibition of spike/hACE2 interaction in vivo. This study presents an innovative in vivo biosensing model to non-invasively quantify virus–host interactions and evaluate the efficacy of spike-targeted antiviral agents under physiologically relevant conditions. [Display omitted] • In vivo NanoBiT pseudoviral biosensor monitored spike–hACE2 binding in real time. • LgBiT-hACE2 mouse model enabled non-invasive tracking of viral entry. • BA.2-SmBiT spike pseudovirus simulated SARS-CoV-2 spike-mediated infection. • NanoBiT bioluminescence imaging enhanced longitudinal infection tracking. • Baicalein and baicalin significantly inhibited spike/hACE2 interaction. [ABSTRACT FROM AUTHOR]
ISSN:00032670
DOI:10.1016/j.aca.2025.344623