Generation and Characterization of a Nanobody Against SARS-CoV

Virol Sin. 2021 Dec;36(6):1484-1491. doi: 10.1007/s12250-021-00436-1. Epub 2021 Aug 17.

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.

Keywords: Nanobody; Neutralizing antibody; Receptor-binding domain (RBD); Severe acute respiratory syndrome coronavirus (SARS-CoV).

MeSH terms

  • Antibodies, Neutralizing
  • Antibodies, Viral / metabolism
  • COVID-19*
  • Escherichia coli / metabolism
  • Humans
  • Leukocytes, Mononuclear / metabolism
  • Protein Binding
  • SARS-CoV-2
  • Severe acute respiratory syndrome-related coronavirus* / metabolism
  • Spike Glycoprotein, Coronavirus / metabolism

Substances

  • Antibodies, Neutralizing
  • Antibodies, Viral
  • Spike Glycoprotein, Coronavirus