Generation of a bivalent dengue virus-like particle targeting DENV-1 and DENV-2 in Pichia pastoris

Shinta Kusumawardani; Muhammad Yusuf; Yosua Yosua; Elza Nurrachmanita Syafrudin; Umi Baroroh; Neni Nurainy; Toto Subroto

doi:10.22146/ijbiotech.114770

Generation of a bivalent dengue virus-like particle targeting DENV-1 and DENV-2 in Pichia pastoris

https://doi.org/10.22146/ijbiotech.114770

Shinta Kusumawardani⁽¹⁾, Muhammad Yusuf⁽²⁾, Yosua Yosua⁽³⁾, Elza Nurrachmanita Syafrudin⁽⁴⁾, Umi Baroroh⁽⁵⁾, Neni Nurainy⁽⁶⁾, Toto Subroto^(7*)

(1) Doctor of Biotechnology Program, Graduated School, Universitas Padjadjaran, West Java, Indonesia; PT. Bio Farma (Persero), Bandung, West, Java, Indonesia
(2) Department of Chemistry, Faculty of Mathematics and Natural Science, Universitas Padjadjaran, West Java, Indonesia; Research Center for Molecular Biotechnology and Bioinformatics, Universitas Padjadjaran, West Java, Indonesia
(3) Research Center for Molecular Biotechnology and Bioinformatics, Universitas Padjadjaran, West Java, Indonesia
(4) Research Center for Molecular Biotechnology and Bioinformatics, Universitas Padjadjaran, West Java, Indonesia
(5) Research Center for Molecular Biotechnology and Bioinformatics, Universitas Padjadjaran, West Java, Indonesia; Department of Biotechnology, Indonesian School of Pharmacy, Bandung, West Java, Indonesia
(6) PT. Bio Farma (Persero), Bandung, West, Java, Indonesia; Department of Biology Pharmacy, Faculty of Pharmacy, Universitas Padjadjaran, West Java, Indonesia
(7) Department of Chemistry, Faculty of Mathematics and Natural Science, Universitas Padjadjaran, West Java, Indonesia; Research Center for Molecular Biotechnology and Bioinformatics, Universitas Padjadjaran, West Java, Indonesia
(*) Corresponding Author

Abstract

Dengue remains a major global health concern, with current licensed vaccines offering variable protection across the four virus serotypes and posing safety considerations related to antibody-dependent enhancement (ADE). To contribute toward improved vaccine design, we developed a bivalent virus-like particle (VLP) targeting DENV-1 and DENV-2, the two most prevalent serotypes in Indonesia. Consensus envelope (CE) protein sequences from Indonesian DENV-1 and DENV-2 strains were codon-optimized and co-expressed in Pichia pastoris GS115 using the AOX1 promoter. The expressed proteins were solubilized from membrane fractions under denaturing conditions, purified via Ni²⁺-affinity chromatography, and subjected to urea gradient dialysis for VLP assembly. Transmission electron microscopy confirmed the formation of spherical VLPs. Western blot analysis confirmed expression and purification via anti-His detection, and supported antigenic integrity of the envelope proteins as recognized by anti-dengue E antibodies. The study demonstrates the feasibility of using P. pastoris for chimeric dengue VLP production based on regionally relevant strains. The findings support the development of serotype-focused VLP platforms as modular components in the development of future tetravalent dengue vaccines.

Keywords

Dengue virus; DENV-1/DENV-2; Envelope protein; Pichia pastoris; Virus-like particle

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DOI: https://doi.org/10.22146/ijbiotech.114770