Bi-paratopic and multivalent VH domains block ACE2 binding and neutralize SARS-CoV-2

QCRG Structural Biology Consortium, CryoEM grid freezing/collection team, CryoEM data processing team, Mammalian cell expression team, Protein purification team, Crystallography team, Bacterial expression team, Infrastructure team, Leadership team

Research output: Contribution to journalArticlepeer-review


Neutralizing agents against SARS-CoV-2 are urgently needed for the treatment and prophylaxis of COVID-19. Here, we present a strategy to rapidly identify and assemble synthetic human variable heavy (VH) domains toward neutralizing epitopes. We constructed a VH-phage library and targeted the angiotensin-converting enzyme 2 (ACE2) binding interface of the SARS-CoV-2 Spike receptor-binding domain (Spike-RBD). Using a masked selection approach, we identified VH binders to two non-overlapping epitopes and further assembled these into multivalent and bi-paratopic formats. These VH constructs showed increased affinity to Spike (up to 600-fold) and neutralization potency (up to 1,400-fold) on pseudotyped SARS-CoV-2 virus when compared to standalone VH domains. The most potent binder, a trivalent VH, neutralized authentic SARS-CoV-2 with a half-maximal inhibitory concentration (IC50) of 4.0 nM (180 ng ml−1). A cryo-EM structure of the trivalent VH bound to Spike shows each VH domain engaging an RBD at the ACE2 binding site, confirming our original design strategy. [Figure not available: see fulltext.]

Original languageEnglish (US)
Pages (from-to)113-121
Number of pages9
JournalNature chemical biology
Issue number1
StatePublished - Jan 2021
Externally publishedYes

ASJC Scopus subject areas

  • Molecular Biology
  • Cell Biology


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