About This Project

Dengue fever is a major health crisis in tropical regions like Indonesia, with Yogyakarta reporting 2,891 cases in 2024, mostly from DENV2 and DENV3 strains (Yogyakarta Health Office, 2024). We address this through a chimeric vaccine targeting both strains using genetic modifications. Our innovative approach uses tobacco plants with a cost-effective genetic promoter, enabling affordable large-scale production and improving vaccine access in endemic regions to protect vulnerable communities.

Ask the Scientists

What is the context of this research?

Our research advances theoretical dengue vaccine designs into practical laboratory testing. We identified a promising chimeric candidate combining DENV-2 and DENV-3 envelope domain III (EDIII) proteins, showing enhanced receptor binding affinity to human cells. To produce this vaccine affordably, we engineer a tobacco plant expression system using a salt-inducible promoter, which activates gene expression under salt stress (Yarra et al., 2012), and an improved INPACT (In Plant Activation) system enabling high-level, controllable protein production (Dugdale et al., 2013). We introduce this construct via Agrobacterium tumefaciens-mediated transformation, a common method for stable gene delivery in plants (Horsch et al., 1985). This integrated, cost-effective platform could transform vaccine accessibility in endemic regions, potentially protecting millions vulnerable to severe dengue disease.

What is the significance of this project?

The significance of this project lies in its innovative approach to using *Nicotiana tabacum*, a tobacco plant widely cultivated in Indonesia, as a host for protein production. By leveraging plant molecular farming, we aim to demonstrate the feasibility of this system for high-yield protein expression, which could be pivotal for vaccine production. Indonesia, with its extensive agricultural resources, holds significant potential to adopt *Nicotiana tabacum* as a new chassis for biopharmaceutical production. Plant molecular farming is renowned for its high yields due to the large biomass of plants, making it an attractive and cost-effective alternative to traditional expression systems. This project presents a proof of concept that not only enhances vaccine production but also positions Indonesia as a key player in the emerging field of plant-based biotechnology.

What are the goals of the project?

This project aims to establish a proof of concept for using Indonesian-grown Nicotiana tabacum as a biomanufacturing platform for producing dengue virus serotype 2 (DENV2) antigens. We are developing protocols to agroinfiltrate 200 plants with our chimeric DENV2 construct, followed by salt-induced protein expression using a modified INPACT system. After extraction, recombinant proteins will be purified using affinity chromatography and validated for their structural integrity and binding efficacy specific to DENV2. The purified antigen will be tested in cell line model DC2.4 and HEK293 cells expressing DENV2 receptors to evaluate immune activation markers and antigen binding through ELISA and Western Blotting. This proof-of-concept study aims to demonstrate the feasibility of using plant-based systems for localized, cost-effective dengue vaccine production, laying the groundwork for future development.