The SpACE-CCM: a facile and versatile cell culture medium-based biosensor for detection of SARS-CoV-2 spike-ACE2 interaction = SARS-CoV-2 Spike-ACE2 단백질간의 상호작용을 이용한 바이오센서 개발
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- The SpACE-CCM: a facile and versatile cell culture medium-based biosensor for detection of SARS-CoV-2 spike-ACE2 interaction = SARS-CoV-2 Spike-ACE2 단백질간의 상호작용을 이용한 바이오센서 개발
- Youngwook Ham; N C Cho; Daeyong; J H Kim; Min Ju Jo; Min Seon Jeong; B Y Pak; S Lee; Mi-Kyung Lee; Seung-Wook Chi; Tae-Don Kim; N C Jeong; Sungchan Cho
- Bibliographic Citation
- Biosensors & Bioelectronics, vol. 227, pp. 115169-115169
- Publication Year
- The COVID-19 pandemic is an ongoing global public health threat. COVID-19 is caused by severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) infection, and binding of the SARS-CoV-2 spike to its receptor, angiotensin-converting enzyme 2 (ACE2), on host cells is critical for viral infection. Here, we developed a luminescent biosensor that readily detects interactions of the spike receptor-binding domain (RBD) and ACE2 in cell culture medium (‘SpACE-CCM’), which was based on bimolecular complementation of the split nanoluciferase-fused spike RBD and ectodomain of ACE2 and further engineered to be efficiently secreted from cells by adding a heterologous secretory signal peptide (SSP). Screening of various SSPs identified ‘interferon-α+alanine-aspartate’ as the SSP that induced the highest activity. The SpACE-CCM biosensor was validated by observing a marked reduction of the activity caused by interaction-defective mutations or in the presence of neutralizing antibodies, recombinant decoy proteins, or peptides. Importantly, the SpACE-CCM biosensor responded well in assay-validating conditions compared with conventional cell lysate-based NanoLuc Binary Technology, indicating its advantage. We further demonstrated the biosensor's versatility by quantitatively detecting neutralizing activity in blood samples from COVID-19 patients and vaccinated individuals, discovering a small molecule interfering with the spike RBD-ACE2 interaction through high-throughput screening, and assessing the cross-reactivity of neutralizing antibodies against SARS-CoV-2 variants. Because the SpACE-CCM is a facile and rapid one-step reaction biosensor that aptly recapitulates the native spike-ACE2 interaction, it would be advantageous in many experimental and clinical applications associated with this interaction.
- COVID-19SARS-CoV-2Spike receptor-binding domainACE2Protein-protein interactionCell culture medium-based biosensor
- Appears in Collections:
- Critical Diseases Diagnostics Convergence Research Center > 1. Journal Articles
Division of Biomedical Research > 1. Journal Articles
Division of Biomedical Research > Immunotherapy Research Center > 1. Journal Articles
Ochang Branch Institute > Nucleic Acid Therapeutics Research Center > 1. Journal Articles
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