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TitleUnconventional linkers facilitate potent stabilized coronavirus stem antibody responses following nanoparticle vaccination.
Journal, issue, pagesbioRxiv, Year 2025
Publish dateNov 19, 2025
AuthorsChristian K O Dzuvor / Sydney P Moak / Lindsay R McManus / Abigail E Dzordzorme / Lamount R Evanson / Taewoo Kim / Abigail Thomas / Olayimika Adeyemi / Valerie Foley / Aditi Limaye / Ryan P McNamara / Kizzmekia S Corbett-Helaire /
PubMed AbstractVaccine technologies that protect against a range of related pathogens within viral families, such as human immunodeficiency virus (HIV), influenza, and coronaviruses (CoVs) represent the future of ...Vaccine technologies that protect against a range of related pathogens within viral families, such as human immunodeficiency virus (HIV), influenza, and coronaviruses (CoVs) represent the future of viral vaccine development. Towards developing broad-spectrum CoV and influenza vaccines, we and others previously designed and evaluated CoV and influenza stem antigens; but these elicited relatively weak and sub-neutralizing antibody (Ab) responses. Multivalent antigen display on nanoparticles (NPs) is an established strategy to enhance and shape immunogenicity. However, one facet of NP vaccines has been largely overlooked: the indispensable linker segment between the antigen and NP core. Here, we introduce -designed rigid (L2) and rarely used long flexible (L6) linkers to optimally display antigens on NPs, target occluded epitopes, and enhance cross-reactive Ab responses, using prefusion-stabilized Middle East respiratory syndrome coronavirus (MERS-CoV) spike (S-2P) and stem (SS) antigens as prototype antigens. Antigenic characterization of L2-NPs confirmed enhanced Ab binding and exposure of cross-reactive epitopes compared with L6-NPs and soluble antigens. Immunization with SS-L2-NPs elicited broader, more potent cross-reactive Ab responses across the seven human-infecting CoVs and pandemic threat WIV1-CoV, whereas SS-L6-NPs induced stronger neutralizing Ab responses against MERS-CoV, SARS-CoV-2, and WIV1-CoV. Ab competition and systems serology analyses revealed that SS-L2-NPs elicit robust Fc-mediated effector functions. By improving CoV-targeting Ab functionality, these linker approaches have the potential to confer broad-spectrum CoV protection and represent a promising strategy against hypervariable influenza and HIV viruses - as well as other broad viral families with pandemic potential.
External linksbioRxiv / PubMed:41332588 / PubMed Central
MethodsEM (single particle)
Resolution4.22 - 6.25 Å
Structure data

EMDB-73668: Structure of SS-L2-FerritinNP
Method: EM (single particle) / Resolution: 6.25 Å

EMDB-73671: Structure of SS-L2-LuSNP
Method: EM (single particle) / Resolution: 4.22 Å

Source
  • Mammalian expression vector Flag-MCS-pcDNA3.1 (others)

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