tetano
Editor, Senior Moderator
Front Immunol
. 2026 Jan 12:16:1726449.
doi: 10.3389/fimmu.2025.1726449. eCollection 2025.
Susceptibility of broad reactivity nanobodies to resistance mutations in the S2 domain of SARS-CoV-2 predicted by yeast display deep mutational scanning
Christina R Ball[SUP] 1 [/SUP], Walter Ramage[SUP] 1 [/SUP], Ryan Mate[SUP] 2 [/SUP], Simon E Hufton[SUP] 1 [/SUP]
Affiliations
Introduction: The rapid evolution of SARS-CoV-2 has led to the erosion of vaccine induced serum neutralization and monoclonal antibody efficacy. As such, interest is inevitably moving towards more conserved regions of the SARS-CoV-2 spike protein like the S2 domain. Resistance mutations continue to be a major obstacle for the development of antivirals and vaccines which target the RBD but what extent these will be a problem for S2 binding antibodies is not known.
Methods: We have developed a yeast display deep scanning mutagenesis platform which allows an unbiased prospective assessment of millions of single and double mutations for their effects on antibody binding to the S2 domain.
Results: We have compared the mutational resistance of a panel of five nanobodies mapped to four distinct non-competing epitopes within the conserved fusion peptide, stem helix and heptad repeat 2 elements of the S2 domain. Yeast display deep mutational scanning predicted reduced binding of C303, G223, G225, and G142 to naturally occurring resistance mutations which were experimentally confirmed on SARS-CoV-2 variants.
Discussion: Our study shows that resistance mutations in conserved elements of the S2 domain may still pose a challenge to the development of monoclonal antibodies and subunit vaccines.
Keywords: S2 domain; SARS-CoV-2; deep mutational scanning; nanobody; pandemic; yeast display.
. 2026 Jan 12:16:1726449.
doi: 10.3389/fimmu.2025.1726449. eCollection 2025.
Susceptibility of broad reactivity nanobodies to resistance mutations in the S2 domain of SARS-CoV-2 predicted by yeast display deep mutational scanning
Christina R Ball[SUP] 1 [/SUP], Walter Ramage[SUP] 1 [/SUP], Ryan Mate[SUP] 2 [/SUP], Simon E Hufton[SUP] 1 [/SUP]
Affiliations
- PMID: 41601659
- PMCID: PMC12832929
- DOI: 10.3389/fimmu.2025.1726449
Introduction: The rapid evolution of SARS-CoV-2 has led to the erosion of vaccine induced serum neutralization and monoclonal antibody efficacy. As such, interest is inevitably moving towards more conserved regions of the SARS-CoV-2 spike protein like the S2 domain. Resistance mutations continue to be a major obstacle for the development of antivirals and vaccines which target the RBD but what extent these will be a problem for S2 binding antibodies is not known.
Methods: We have developed a yeast display deep scanning mutagenesis platform which allows an unbiased prospective assessment of millions of single and double mutations for their effects on antibody binding to the S2 domain.
Results: We have compared the mutational resistance of a panel of five nanobodies mapped to four distinct non-competing epitopes within the conserved fusion peptide, stem helix and heptad repeat 2 elements of the S2 domain. Yeast display deep mutational scanning predicted reduced binding of C303, G223, G225, and G142 to naturally occurring resistance mutations which were experimentally confirmed on SARS-CoV-2 variants.
Discussion: Our study shows that resistance mutations in conserved elements of the S2 domain may still pose a challenge to the development of monoclonal antibodies and subunit vaccines.
Keywords: S2 domain; SARS-CoV-2; deep mutational scanning; nanobody; pandemic; yeast display.