Tuning Antigen–Adjuvant Interactions by Modulating the Physicochemical Properties of Aluminum Hydroxide Nanoparticles for Improved Antigen Stability

Adjuvants are chemical substances used in vaccines to enhance immunogenicity. Among them, aluminum-based nanoparticles are some of the oldest and most widely employed adjuvants in vaccine formulations. A key function of aluminum adjuvants is thought to involve acting as an antigen depot, enabling slow antigen release and providing sufficient time for effective immune activation. Therefore, understanding antigen–adjuvant interactions is essential, as these interactions influence antigen stability, release kinetics, and overall vaccine performance. In this study, we investigated how the physicochemical properties of aluminum hydroxide nanoparticles modulate antigen–protein interactions and affect protein stability. Nanoparticles synthesized under acidic (pH ≈ 5.0) to near-neutral (pH ≈ 7.1) conditions exhibited lower crystallinity, reduced hydroxyl density, and higher interfacial hydration, whereas those prepared under basic conditions (pH ≈ 9.0) displayed increased crystallinity, enriched surface hydroxyl groups, and markedly reduced hydration. Antigen proteins bound to low-crystallinity aluminum hydroxide nanoparticles showed improved thermal stability, while those associated with highly crystalline nanoparticles exhibited reduced thermal stability. Complementary ITC study further suggests that these stability differences are accompanied by changes in their interaction behavior. These findings indicate that the structural and interfacial properties of aluminum hydroxide nanoparticles strongly influence their interactions with antigen proteins and the resulting physical stability. Together, our results demonstrate that the balance among crystallinity, hydroxyl organization, and interfacial hydration governs the thermal behavior of antigen proteins adsorbed onto aluminum hydroxide. This work provides a rational design principle for engineering aluminum-based adjuvants that optimize antigen–protein stability in vaccine formulations.