Enhancing the sensitivity of nanobodies through covalent and non-covalent polymerization.

ABSTRACT: The precise identification of disease using antibodies is crucial for guiding therapeutic interventions. Among these binding proteins, the nanobody is the smallest known antigen-binding fragment. This study aimed to enhance antigen detection sensitivity by integrating covalent dimerization and non-covalent heptamerization strategies to increase the density of nanobodies for improved antigen capture. First, we synthesized a SpyTag-Catcher-fused nanobody and a heptamerization protein using a prokaryotic expression system. Western blot was used to preliminarily verify the nanobody's antigen-binding capability. Surface Plasmon Resonance (SPR) analysis revealed that the enhanced immunoactivity of the nanobody polymerization resulted from their increased affinity. Finally, we employed these nano

SUBMITTER: Wang F 

PROVIDER: S-EPMC12814022 | biostudies-literature | 2026 Jan

REPOSITORIES: biostudies-literature