Zika virus-like particles (VLPs) produced in insect cells

Frontiers in Pharmacology

Pharmacology

Eutimio Gustavo Fernández Núñez¹  Simone Gonçalves Silva Jared²  Marta Maria Antoniazzi²  Soraia Attie Calil Jorge³  Renata Gois de Mello³  Luis Giovani Oliveira Guardalini³  Renato Mancini Astray³  Thaissa Consoni Bernardino³ 

[1] Grupo de Engenharia de Bioprocessos, Escola de Artes, Ciências e Humanidades (EACH), Universidade de São Paulo, São Paulo, SP, Brazil;Laboratório de Biologia Estrutural, Instituto Butantan, São Paulo, SP, Brazil;Laboratório de Biotecnologia Viral, Instituto Butantan, São Paulo, SP, Brazil;

关键词: virus-like particles;    Zika virus;    recombinant baculovirus;    Sf9 cells;    insect cells;   

DOI  :  10.3389/fphar.2023.1181566

 received in 2023-03-07, accepted in 2023-05-30,  发布年份 2023

来源: Frontiers

【 摘 要 】

Introdutcion: The Zika virus (ZIKV) infections are a healthcare concern mostly in the Americas, Africa, and Asia but have increased its endemicity area beyond these geographical regions. Due to the advances in infections by Zika virus, it is imperative to develop diagnostic and preventive tools against this viral agent. Virus-like particles (VLPs) appear as a suitable approach for use as antiviral vaccines.Methods: In this work, a methodology was established to produce virus-like particles containing the structural proteins, C, prM, and E of Zika virus produced in insect cells using the gene expression system derived from baculovirus. The vector pFast- CprME -ZIKV was constructed containing the gene sequences of Zika virus structural proteins and it was used to generate the recombinant bacmids (Bac- CprME -ZIKV) through transformation into DH10BacTM cells. The Bac- CprME -ZIKV was transfected in Spodoptera frugiperda (Sf9) insect cells and batches of BV- CprME -ZIKV were obtained by infection assays using a multiplicity of infection of 2. The Sf9 cells were infected, and the supernatant was collected 96 h post-infection. The expression of the CprME -ZIKV protein on the cell surface could be observed by immunochemical assays. To concentrate and purify virus-like particles, the sucrose and iodixanol gradients were evaluated, and the correct CprME -ZIKV proteins’ conformation was evaluated by the Western blot assay. The virus-like particles were also analyzed and characterized by transmission electron microscopy.Results and discussion: Spherical structures like the native Zika virus from 50 to 65 nm containing the CprME -ZIKV proteins on their surface were observed in micrographs. The results obtained can be useful in the development path for a vaccine candidate against Zika virus.

【 授权许可】

Unknown   
Copyright © 2023 de Mello, Bernardino, Guardalini, Astray, Antoniazzi, Jared, Núñez and Jorge.