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.

中文翻译：

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基于酪氨酸的肽抗原交联以产生具有增强免疫原性的纳米簇：使用甲型流感的保守 M2e 肽进行演示

描述了利用基于酪氨酸-酪氨酸交联反应的方法从可溶性肽分子产生纳米簇 (NC) 的方法。在肽分子的末端引入包含组氨酸和酪氨酸残基的反应性标签。交联反应导致标签内产生二酪氨酸键，从而产生肽 NC。我们表明，反应性标签必须存在于肽的“N”和“C”末端，才能形成簇。此外，系统地表征了交联反应，以显示反应条件对最终簇直径的重要性，使我们能够生成各种尺寸的 NC。为了证明肽簇的免疫原性潜力，我们选择研究保守的流感肽，M2e，作为抗原。使用交联反应配制 M2e NC。我们展示了集群在 BALB/c 小鼠中以剂量、大小和频率依赖的方式产生保护性免疫的能力，以对抗致命的甲型流感病毒。总之，所提供的数据表明，这种新的簇形成技术可以以简单和可控的方式产生高度免疫原性肽 NC。