Synthesis of Chirally Chimeric Protein Nanoparticle Vaccines via Mirror-Image Spy Chemistry.

ABSTRACT: Mirror-image proteins and nucleic acids exhibit remarkable biostability and bioorthogonality, offering a promising strategy to enhance the longevity of biological therapeutics. Here, we present a modular approach for constructing complex protein architectures that integrate both natural-chirality (L-) and mirror-image (D-) motifs. Key to this strategy is the development of D-SpyStapler-a chemically synthesized, chirally inverted ligase that enables the efficient conjugation of D-SpyTag and D-BDTag. By exploiting the achirality of glycine (Gly), we used L-sortase to covalently link D-peptides (e.g., SpyTag or BDTag, bearing an N-terminal poly-Gly motif) to L-proteins (e.g., GFP, VLP-forming Mi3, or antigens) containing a C-terminal LPXTG sorting signal. The resulting D-SpyTagged and D-BDTag

SUBMITTER: Kou S 

PROVIDER: S-EPMC12363629 | biostudies-literature | 2025 Aug

REPOSITORIES: biostudies-literature