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Generation and Characterization of a Nanobody Against SARS-CoV

Jiangfan Li, Lei He, Yong‐Qiang Deng, Shuhui Qi, Yuehong Chen, Xiaolu Zhang, Shixiong Hu, Ruiwen Fan, Guangyu Zhao, Cheng‐Feng Qin

2021Virologica Sinica19 citationsDOIOpen Access PDF

Abstract

The sudden emergence of severe acute respiratory syndrome coronavirus (SARS-CoV) has caused global panic in 2003, and the risk of SARS-CoV outbreak still exists. However, no specific antiviral drug or vaccine is available; thus, the development of therapeutic antibodies against SARS-CoV is needed. In this study, a nanobody phage-displayed library was constructed from peripheral blood mononuclear cells of alpacas immunized with the recombinant receptor-binding domain (RBD) of SARS-CoV. Four positive clones were selected after four rounds of bio-panning and subjected to recombinant expression in E. coli. Further biological identification demonstrated that one of the nanobodies, S14, showed high affinity to SARS-CoV RBD and potent neutralization activity at the picomole level against SARS-CoV pseudovirus. A competitive inhibition assay showed that S14 blocked the binding of SARS-CoV RBD to either soluble or cell-expressed angiotensin-converting enzyme 2 (ACE2). In summary, we developed a novel nanobody targeting SARS-CoV RBD, which might be useful for the development of therapeutics against SARS.

Topics & Concepts

Recombinant DNAPanning (audio)VirologyAntibodyPeripheral blood mononuclear cellSevere acute respiratory syndrome coronavirus 2 (SARS-CoV-2)Phage displaySevere acute respiratory syndromeBiologyCoronavirusNeutralizationGlycoproteinPeptide libraryCoronavirus disease 2019 (COVID-19)ChemistryMolecular biologyImmunologyVirusMedicineGenePeptide sequenceIn vitroBiochemistryInfectious disease (medical specialty)DiseaseZoomPathologyLens (geology)PaleontologySARS-CoV-2 and COVID-19 ResearchMonoclonal and Polyclonal Antibodies ResearchBacteriophages and microbial interactions
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