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(Referência obtida automaticamente do Web of Science, por meio da informação sobre o financiamento pela FAPESP e o número do processo correspondente, incluída na publicação pelos autores.)

Production of Rabies VLPs in Insect Cells by Two Monocistronic Baculoviruses Approach

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Autor(es):
Bernardino, Thaissa Consoni [1] ; Astray, Renato Mancini [1] ; Pereira, Carlos Augusto [1] ; Boldorini, Vera Lucia [1] ; Antoniazzi, Marta Maria [2] ; Silva Jared, Simone Goncalves [2] ; Fernandez Nunez, Eutimio Gustavo [3] ; Calil Jorge, Soraia Attie [1]
Número total de Autores: 8
Afiliação do(s) autor(es):
[1] Inst Butantan, Lab Biotecnol Viral, Av Vital Brasil 1500, BR-05503900 Sao Paulo - Brazil
[2] Inst Butantan, Lab Biol Estrutural, Sao Paulo, SP - Brazil
[3] Univ Sao Paulo, Escola Artes Ciencias & Humanidades EACH, Grp Engn Bioproc, Sao Paulo, SP - Brazil
Número total de Afiliações: 3
Tipo de documento: Artigo Científico
Fonte: MOLECULAR BIOTECHNOLOGY; v. 63, n. 11 JUL 2021.
Citações Web of Science: 0
Resumo

Rabies is an ancient zoonotic disease that still causes the death of over 59,000 people worldwide each year. The rabies lyssavirus encodes five proteins, including the envelope glycoprotein and the matrix protein. RVGP is the only protein exposed on the surface of viral particle, and it can induce immune response with neutralizing antibody formation. RVM has the ability to assist with production process of virus-like particles. VLPs were produced in recombinant baculovirus system. In this work, two recombinant baculoviruses carrying the RVGP and RVM genes were constructed. From the infection and coinfection assays, we standardized the best multiplicity of infection and the best harvest time. Cell supernatants were collected, concentrated, and purified by sucrose gradient. Each step was used for protein detection through immunoassays. Sucrose gradient analysis enabled to verify the separation of VLPs from rBV. Through the negative contrast technique, we visualized structures resembling rabies VLPs produced in insect cells and rBV in the different fractions of the sucrose gradient. Using ELISA to measure total RVGP, the recovery efficiency of VLPs at each stage of the purification process was verified. Thus, these results encourage further studies to confirm whether rabies VLPs are a promising candidate for a veterinary rabies vaccine. (AU)

Processo FAPESP: 09/08038-1 - Estudo de expressão gênica empregando pseudopartículas virais
Beneficiário:Soraia Attie Calil Jorge
Modalidade de apoio: Auxílio à Pesquisa - Regular