(T1130-02-08) An Oleic Acid-Conjugated Vaccine Antigen for Enhanced Physicochemical Properties and Thermal Stability via Self-Assembled Nanonization

Purpose: To improve the physicochemical properties and thermal stability of virus vaccine antigen (Heg) via self-assembled nanonization of oleic acid conjugated Heg (HOC) to form nanoparticles (HON.

Methods: HOC was synthesized via 1-Ethyl-3(3-dimethylaminopropyl) carbodiimide (EDC)/N-hydroxysuccinimide (NHS) reaction. The synthesis of new amphiphilic conjugates was identified through matrix-assisted laser desorption ionization time-of-flight mass spectroscopy (MALDI-TOF-MS). Solvent evaporation method was utilized to prepare HON from HOC. The effect of solvent types on the nanonization of HOC was evaluated using ethanol, methanol, and tetrahydrofuran (THF) at the solvent/water ratio of 1:5 (v/v). The morphology of HON was analyzed by SEM and TEM. Thermal stability of Heg and HON was investigated at 40℃ for 7days using size exclusion chromatography (SEC) HPLC, MALDI-TOF-MS and sodium dodecyl-sulfate polyacrylamide gel electrophoresis (SDS-PAGE) methods.

Results: The conjugation of oleic acid to vaccine antigen was successfully confirmed with an increase of molecular weight of Heg from around 85.8 to 90.3 kDa. It was confirmed that the solvent types had an impact on the nanonization process of HOC and nanoparticle sizes as visualized by SEM and TEM. The thermal stability of HON highly increased without showing any degradation or new peaks at 1, 3 and 7 days in SDS-PAGE unlike the free vaccine antigen. In a quantitative analysis of SEC HPLC, about 20% and 50% of Heg decreased at 1 and 3 days while the amount of HON was almost unchanged and over 90% HOC remained for 3days.

Conclusion: The fattigation platform by conjugating fatty acid to vaccine antigen could provide a new strategy to improve thermal stability and physicochemical property of model vaccine antigen.

Acknowledgements:Supported by a grant from the Korea Health Technology R&D Project through the Korea Health Industry Development Institute (KHIDI), funded by the Ministry of Health & Welfare, The Republic of Korea (Grant No.: HV22C0222).