Presentation Information
[4FMBS-03]Enzymatic post-modification of self-assembling peptides for biochemical applications
○Rie Wakabayashi1, Noriho Kamiya1, Masahiro Goto1 (1. Kyushu University (Japan))
Keywords:
Self-assembly,Peptides,Enzymatic reaction,Protein,vaccines
[Purpose]
This study aimed to fabricate protein-integrated materials based on self-assembling peptides (SAPs) for potential biochemical applications including vaccines. SAP-based supramolecular materials offer design diversity, adaptability to environments, and inherent biocompatibility. However, controlled integration of proteins while preserving both supramolecular structures and protein functions remains challenging. Through molecular design and post-assembly functionalization of SAPs, hierarchical structures and assembly properties can be modulated. Taking advantages of these features, we investigated the bio-functions, such as cellular interactions, of protein-integrated SAP materials.
[Method]
Enzymatic reactions were employed to post-modify self-assembled peptide fibers with proteins. SAPs were designed to possess both self-assembling capability and enzymatic reaction motifs. SAPs were first allowed to self-assemble into nanofibers, followed by enzymatic conjugation with green fluorescent protein, GFP, as a model antigenic protein. The effects of integration of GFPs onto the SAP fibers on in vitro delivery to dendritic cells and in vivo vaccine efficacy in mice were evaluated.
[Results]
Enzymatic reactions enabled the integration of multiple proteins onto SAP fibers without compromising fiber structures or protein structure/function.(1) When conjugated to SAP fibers, GFP exhibited enhanced internalization into immune cells in an SAP design-dependent manner. Furthermore, in vivo vaccine efficacy, evaluated by the serum antibody titers, was significantly enhanced by the integration onto the SAP fibers.(2)
[Consideration]
Enzymatic reactions were performed under the conditions compatible with both proteins and SAPs (< 1 h at 37°C, pH 7.0), thereby preserving their structures and functions. The physicochemical properties of SAP fibers, such as hydrophobicity and surface charge, strongly influenced cellular interactions and internalization of the conjugated GFP. Moreover, in vivo vaccine efficacy of the antigenic proteins was enhanced by the integration onto SAP fibers, suggesting that this enzymatic post-modification strategy is a useful approach for controlling the spatial presentation and physicochemical environment to modulate and enhance protein bio-functions.
[Conclusion]
SAPs with enzymatic reactivity successfully enabled the integration of proteins onto assembled nanofibers, leading to enhanced bio-functions, including intracellular delivery and in vivo vaccine efficacy. This enzymatic post-assembly modification strategy represents a promising approach for functionalizing protein-based biomaterials.
[References]
(1) Wakabayashi et al., Chem. Commun., 55, 640 (2019)
(2) Wakabayashi et al., ACS Appl. Mater. Interfaces, 17, 44240 (2025)
This study aimed to fabricate protein-integrated materials based on self-assembling peptides (SAPs) for potential biochemical applications including vaccines. SAP-based supramolecular materials offer design diversity, adaptability to environments, and inherent biocompatibility. However, controlled integration of proteins while preserving both supramolecular structures and protein functions remains challenging. Through molecular design and post-assembly functionalization of SAPs, hierarchical structures and assembly properties can be modulated. Taking advantages of these features, we investigated the bio-functions, such as cellular interactions, of protein-integrated SAP materials.
[Method]
Enzymatic reactions were employed to post-modify self-assembled peptide fibers with proteins. SAPs were designed to possess both self-assembling capability and enzymatic reaction motifs. SAPs were first allowed to self-assemble into nanofibers, followed by enzymatic conjugation with green fluorescent protein, GFP, as a model antigenic protein. The effects of integration of GFPs onto the SAP fibers on in vitro delivery to dendritic cells and in vivo vaccine efficacy in mice were evaluated.
[Results]
Enzymatic reactions enabled the integration of multiple proteins onto SAP fibers without compromising fiber structures or protein structure/function.(1) When conjugated to SAP fibers, GFP exhibited enhanced internalization into immune cells in an SAP design-dependent manner. Furthermore, in vivo vaccine efficacy, evaluated by the serum antibody titers, was significantly enhanced by the integration onto the SAP fibers.(2)
[Consideration]
Enzymatic reactions were performed under the conditions compatible with both proteins and SAPs (< 1 h at 37°C, pH 7.0), thereby preserving their structures and functions. The physicochemical properties of SAP fibers, such as hydrophobicity and surface charge, strongly influenced cellular interactions and internalization of the conjugated GFP. Moreover, in vivo vaccine efficacy of the antigenic proteins was enhanced by the integration onto SAP fibers, suggesting that this enzymatic post-modification strategy is a useful approach for controlling the spatial presentation and physicochemical environment to modulate and enhance protein bio-functions.
[Conclusion]
SAPs with enzymatic reactivity successfully enabled the integration of proteins onto assembled nanofibers, leading to enhanced bio-functions, including intracellular delivery and in vivo vaccine efficacy. This enzymatic post-assembly modification strategy represents a promising approach for functionalizing protein-based biomaterials.
[References]
(1) Wakabayashi et al., Chem. Commun., 55, 640 (2019)
(2) Wakabayashi et al., ACS Appl. Mater. Interfaces, 17, 44240 (2025)
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