Tyrosine-Based Cross-Linking of Peptide Antigens to
Generate Nanoclusters with Enhanced Immunogenicity: Demonstration
Using the Conserved M2e Peptide of Influenza A
posted on 2021-08-25, 20:14authored byLogan
R. Wilks, Gaurav Joshi, Megan R. Grisham, Harvinder Singh Gill
A method of creating nanoclusters
(NCs) from soluble peptide molecules
is described utilizing an approach based on a tyrosine-tyrosine cross-linking
reaction. A reactive tag comprising histidine and tyrosine residues
was introduced at the termini of the peptide molecules. The cross-linking
reaction led to the creation of dityrosine bonds within the tag, which
allowed for the generation of peptide NCs. We show that it is essential
for the reactive tag to be present at both the “N” and
“C” termini of the peptide for cluster formation to
occur. Additionally, the cross-linking reaction was systematically
characterized to show the importance of reaction conditions on final
cluster diameter, allowing us to generate NCs of various sizes. To
demonstrate the immunogenic potential of the peptide clusters, we
chose to study the conserved influenza peptide, M2e, as the antigen.
M2e NCs were formulated using the cross-linking reaction. We show
the ability of the clusters to generate protective immunity in a dose,
size, and frequency dependent manner against a lethal influenza A
challenge in BALB/c mice. Taken together, the data presented suggest
this new cluster formation technique can generate highly immunogenic
peptide NCs in a simple and controllable manner.