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Pre-Clinical Evaluation of a Novel Nanoemulsion-Based Hepatitis B Mucosal Vaccine

Figure 2

HBsAg interaction with NE droplets.

Zeta potential: (A) The zeta potential was measured for HBsAg (2.5 mg/ml), 20% NE and mixture of HBsAg with NE (HBsAg-NE) in water and PBS. Surface charge is reported in mV units. Particle sizing: Size distribution was measured using a laser diffraction particle-sizer. Analysis of HBsAg alone (B), NE alone (C), and NE with 10 μg/ml of HBsAg (D). Data was processed and analyzed using Fraunhofer optical modeling and number weighted averaging (number %). Single population intensity peaks indicate monodisperse populations of HBsAg (28 nm), NE (349 nm), and HBsAg-NE (335 nm). Calorimetric titration of HBsAg with NE: 25 injections of 1% NE (10 μl/injection) were introduced into a sample cell containing HBsAg (600 μg/ml in PBS) at 30°C. The upper panel shows differences between the sample and reference cell containing PBS. The lower panel shows enthalpy per injection of NE injected versus injection number. An exothermic reaction was measured following the addition of nanoemulsion to an antigen solution. Addition of nanoemulsion to the antigen solution became less energetic as the total concentration of nanoemulsion increased, suggesting that the antigen was being depleted. ΔCp was calculated to be −1.44 [77].

Figure 2

doi: https://doi.org/10.1371/journal.pone.0002954.g002