Proceedings of the 8th Brazilian Congress of Biotechnology

DEVELOPMENT OF VLPS FOR SARS-COV-2 USING INSECT CELL SYSTEM

Published in 21/11/2025 - ISBN: 978-65-272-1865-4

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Paper Title
DEVELOPMENT OF VLPS FOR SARS-COV-2 USING INSECT CELL SYSTEM
Authors
  • Luis Giovani de Oliveira Guardalini
  • Thaissa Consoni Bernardino
  • Wagner Quintilio
  • Eutimio Gustavo Fernández Núñez
  • Soraia Attie Calil Jorge
Modality
Abstract
Subject area
Applications of Biotechnology in Human Health (vaccine and drug development);
Publishing Date
21/11/2025
Country of Publishing
Brazil | Brasil
Language of Publishing
en-US
Paper Page
https://www.even3.com.br/anais/biotecnologia2025/1234340-development-of-vlps-for-sars-cov-2-using-insect-cell-system
ISBN
978-65-272-1865-4
Keywords
Recombinant baculovirus, Virus-like particles (VLPs), Insect cells, SARS-CoV-2
Summary
The SARS-CoV-2 virus, responsible for the COVID-19 pandemic, has posed significant challenges to global public health. In response, the development of safe and effective vaccines has remained a key focus in biomedical research. This study explores the generation of Virus-Like Particles (VLPs) from SARS-CoV-2 through co-infection with recombinant baculoviruses carrying genetic information encoding the virus’s structural proteins. VLPs offer a promising alternative, capable of eliciting a strong immune response without the risk of actual viral infection. The main objective of this work is the production of SARS-CoV-2 VLPs as a potential vaccine candidate. To achieve this, commercial plasmids were acquired and transformed into E. coli to allow site-specific transposition of genetic inserts into bacmids. Following transformation, alkaline lysis was performed to extract the bacmids, which represent the condensed genetic material of the baculovirus. These bacmids were then used to transfect insect cells, leading to the generation of viral stocks of recombinant baculoviruses containing the genes encoding SARS-CoV-2 structural proteins. Bioreactor screening tests were also conducted to determine the optimal combination of baculoviruses for co-infection. This step was crucial to ensure efficient protein expression and to identify combinations most likely to assemble into virus-like structures. Transmission electron microscopy (TEM) was used to characterize the resulting VLPs, while dot blot assays were employed to assess protein expression. After transformation and lysis, bacmid DNA concentrations were obtained in the range of 990 µg/mL for Bac-S, 870 µg/mL for Bac-M, 580 µg/mL for Bac-N, 830 µg/mL for Bac-RBD, and 980 µg/mL for Bac-E. Transfection resulted in baculovirus batches with titers of 10⁶ pfu/mL for first-passage batches, 10⁷ pfu/mL for second-passage, and 10⁸ pfu/mL for third-passage batches. Expression kinetics, measured by dot blot in coinfected samples within the bioreactor, showed that structural protein expression peaked around 63 hours post-coinfection, using the most effective combination identified through TEM analysis. These findings contribute to optimizing conditions for VLP production. TEM micrographs confirmed that co-infection with the baculoviruses BVS:BVN:BVM, at multiplicities of infection of 3:2:1 respectively, yielded the most consistent formation of circular particles with an average diameter of approximately 100 nm. This study demonstrates the feasibility of generating SARS-CoV-2 VLPs, and the results are promising for the development of a vaccine candidate. Efficient VLP production, coupled with the expression of the virus’s structural proteins, represent a critical step toward the creation of an effective COVID-19 vaccine. The work conducted here contributes to the growing body of knowledge essential for rapid responses to future viral emergencies.
Title of the Event
8th Brazilian Biotechnology Congress.
City of the Event
Natal
Title of the Proceedings of the event
Proceedings of the 8th Brazilian Congress of Biotechnology
Name of the Publisher
Even3
Means of Dissemination
Meio Digital

How to cite

GUARDALINI, Luis Giovani de Oliveira et al.. DEVELOPMENT OF VLPS FOR SARS-COV-2 USING INSECT CELL SYSTEM.. In: Proceedings of the 8th Brazilian Congress of Biotechnology. Anais...Natal(RN) Hotel Praia Mar, 2025. Available in: https//www.even3.com.br/anais/biotecnologia2025/1234340-DEVELOPMENT-OF-VLPS-FOR-SARS-COV-2-USING-INSECT-CELL-SYSTEM. Access in: 22/06/2026

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