Optimization of Dengue-EDIII – Hepatitis B-VLP Fusion Antigen Protein Expression in Mammalian Cell Systems and Preliminary Immunogenicity Testing in Mice

Ziza Putri Aisyia Fauzi (Universitas Muslim)
Eka Nur Fatmawati (Akbid Harapan Mulia Ponorogo Akbid Harapan Mulia Ponorogo)
Putri Tri Hartini (Universitas Muhammadiyah Riau)
Yuneka Saristiana (Universitas Kadiri)
Husnunnisa (Universitas Muhammadiyah Riau)

Fundamental and Applied Research in Medicine and Allied Sciences Indonesia
Vol. 1 No. 2 (2025): November, 2025

The development of a safe and effective dengue vaccine remains challenging due to the risk of antibody-dependent enhancement (ADE) associated with imbalanced immune responses. This study aimed to optimize the expression of a dengue virus envelope domain III (EDIII) fusion antigen displayed on Hepatitis B core antigen virus-like particles (VLPs) and to evaluate its preliminary immunogenicity in a murine model. Optimization of antigen design and expression conditions resulted in a substantial increase in VLP production compared with the non-optimized construct. The purified VLPs showed uniform size, proper particle assembly, and preserved antigenic integrity. Immunization of mice induced high titers of EDIII-specific IgG and strong serotype-specific neutralizing antibodies against dengue virus serotype 2, while no antibody-dependent enhancement was detected in vitro. These findings indicate that the optimized EDIII–HBcAg VLP platform is highly immunogenic and demonstrates a favorable safety profile, supporting its potential as a promising subunit vaccine candidate for dengue virus.