A Modular Antigen-Anchoring Strategy for Cellular Nanoparticle Vaccines Against Burkholderia pseudomallei

Abstract

A challenge in vaccine engineering is the lack of adaptable platforms that allow rapid formulation of complex, multiantigen vaccines while preserving antigen stability, orientation, and immunological function. Here, we report a plug-and-play biomimetic nanovaccine platform based on cellular nanoparticles (CNPs) that enables rapid and chemically defined antigen assembly via a modular antigen-anchoring strategy. In this approach, protein and polysaccharide antigens are independently conjugated to lipid anchors through click chemistry and spontaneously inserted into the membrane of preformed CNPs, thereby decoupling antigen functionalization from nanoparticle fabrication and enabling combinatorial design without reengineering the nanocarrier. Using B. pseudomallei antigens (Hcp1 and CPS) as a representative example, we show that the resulting antigen-displayed CNPs exhibit controlled surface presentation, preserved antigen orientation, and strong physicochemical stability. This modular assembly enhances dendritic cell uptake, maturation, and cytokine production and promotes efficient activation of antigen-presenting cells in draining lymph nodes. Importantly, the platform elicits concurrent CD4+ and CD8+ T-cell responses together with high-titer, high-avidity antibody responses and germinal center formation, demonstrating its ability to drive coordinated adaptive immunity. Collectively, this work establishes a versatile and rapidly deployable framework for nanovaccine development.