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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]
Copyright of Analytica Chimica Acta is the property of Elsevier B.V. and its content may not be copied or emailed to multiple sites without the copyright holder's express written permission. Additionally, content may not be used with any artificial intelligence tools or machine learning technologies. However, users may print, download, or email articles for individual use. This abstract may be abridged. No warranty is given about the accuracy of the copy. Users should refer to the original published version of the material for the full abstract. (Copyright applies to all Abstracts.)
Database: Engineering Source
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  Data: 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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  Data: <searchLink fieldCode="AR" term="%22Lin%2C+Cheng-Han%22">Lin, Cheng-Han</searchLink><relatesTo>1</relatesTo> (AUTHOR)<br /><searchLink fieldCode="AR" term="%22Lin%2C+Meng-Wei%22">Lin, Meng-Wei</searchLink><relatesTo>1</relatesTo> (AUTHOR)<br /><searchLink fieldCode="AR" term="%22Yang%2C+Xin-Rui%22">Yang, Xin-Rui</searchLink><relatesTo>1</relatesTo> (AUTHOR)<br /><searchLink fieldCode="AR" term="%22Chiang%2C+Hua-Hsin%22">Chiang, Hua-Hsin</searchLink><relatesTo>1</relatesTo> (AUTHOR)<br /><searchLink fieldCode="AR" term="%22Wu%2C+Ting-Hsuan%22">Wu, Ting-Hsuan</searchLink><relatesTo>1</relatesTo> (AUTHOR)<br /><searchLink fieldCode="AR" term="%22Tsai%2C+Chin-Hung%22">Tsai, Chin-Hung</searchLink><relatesTo>2</relatesTo> (AUTHOR)<br /><searchLink fieldCode="AR" term="%22Lin%2C+Chih-Sheng%22">Lin, Chih-Sheng</searchLink><relatesTo>1,3</relatesTo> (AUTHOR)<i> lincs@nycu.edu.tw</i>
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  Label: Abstract
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  Data: 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]
– Name: AbstractSuppliedCopyright
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  Data: <i>Copyright of Analytica Chimica Acta is the property of Elsevier B.V. and its content may not be copied or emailed to multiple sites without the copyright holder's express written permission. Additionally, content may not be used with any artificial intelligence tools or machine learning technologies. However, users may print, download, or email articles for individual use. This abstract may be abridged. No warranty is given about the accuracy of the copy. Users should refer to the original published version of the material for the full abstract.</i> (Copyright applies to all Abstracts.)
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RecordInfo BibRecord:
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      – Type: doi
        Value: 10.1016/j.aca.2025.344623
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        Text: English
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        PageCount: 1
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      – SubjectFull: SARS-CoV-2
        Type: general
      – SubjectFull: Biosensors
        Type: general
      – SubjectFull: Membrane glycoproteins
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      – SubjectFull: Angiotensin converting enzyme
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      – SubjectFull: Clinical trials
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      – SubjectFull: Bioluminescence
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      – SubjectFull: Real-time computing
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            NameFull: Lin, Cheng-Han
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              Text: Nov2025
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