Development of monoclonal antibodies against prM of Dengue virus 4

Auteurs

DOI :

https://doi.org/10.9771/cmbio.v21i1.44965

Mots-clés :

Anticorpos monoclonais. DENV 4. prM

Résumé

Introduction: dengue is a most common mosquito-borne viral disease in the Americas and tropical countries. Objective: in this work, mice were hyperimmunized   with DENV 4 antigen to produce monoclonal antibodies (mAbs).  Methodology: DENV 4 (GenBank KC806069) was inoculated in C6/36 cell monolayers cultivated in Leibovitz’s 15 medium supplemented with 5% fetal bovine serum and incubated at 28 C. The virus stock was submitted to concentration and ultracentrifugation and stored at -80 C until use (VC DENV 4). Balb/c mice were injected intraperitoneally with 50μg of DENV-4 and successive intraperitoneal injections of 25 μg of VC DENV 4 with Freund’s incomplete adjuvant were performed. The spleen cells were fused to SP2/0 myeloma cells with PEG 1540 and distributed in 96-well microplates with Iscove’s modified medium with Hipoxantina–Aminopterina–Timidina. Hybridoma screening by indirect ELISA showed positive results for six mAbs, and their characterization was performed by Western blotting and Indirect Immunofluorescence (IFI) techniques. Results: the six mAbs showed strong recognition of prM (24/29 kDa), and minor reaction to E protein (66 kDa), E/E protein dimer (105 kDa), and NS1 (49 kDa) protein in two mAbs. The use of mAbs anti-prM as a diagnostic tool using IFI has been demonstrated to detect DENV-4 antigen in infected cells or tissues. Conclusion: DENV 4 generate mAbs with strong reactivity to prM with potential use to confirm the presence of DENV 4 antigen in tissues or infected cells.

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Bibliographies de l'auteur

Adriana de Souza Andrade, Colaboradora do Laboratorio de Virologia 2 andar.

Farmacêutica/Bioquímica, Esp. Microbiologia, Msc. Biotecnologia. Doutora em Microbiologia- Virologia (UFMG) pelaUniversidade Federal de Minas Gerais. 

Gubio Soares Campos, Farmaceutico. UFBA. ICS

Farmaceutico. UFBA. PhD virologia Universidad de Buenos Aires. Coordenador do Laboratorio de virologia 2 andar ICS.

Lilia Ferreira Moura Costa, Professora Associada - Microbiologia Departamento de Ciências da Biointeração Instituto de Ciências da Saúde Universidade Federal da Bahia Salvador - BA Brasil

M.D., PhD. Professora Associada - Microbiologiahttp://lattes.cnpq.br/6672994596587713
Departamento de Ciências da BiointeraçãoInstituto de Ciências da Saúde
Universidade Federal da Bahia

Adriano Figuereido Monte-Alegre, Professor Associado.Departamento de Ciências da Biointeração. Paarasitologia.Instituto de Ciências da SaúdeUniversidade Federal da BahiaSalvador - BABrasil

Professor Associado de Parasitologia da Universidade Federal da Bahia - UFBA, Instituto de Ciencias da Saúde, Departamento de Biointeração (2010). Graduação em Ciências Biologicas (1997), Especialização em Entomologia Médica - USP, Mestrado em Biosciences de lEnvironnement, Chimie et Santé (ênfase em Biologia evolutiva) - Universidade de Provence - Marselha-França (2002) e Doutorado em Parasitologia - Universidade de Montpellier II -França (2006). Área de domínio - Parasitologia, com ênfase em Leishmania (parasita protozoário) e Flebotomos (inseto vetor).

Silvia Inês Sardi, Universidade Federal da Bahia - UFBA

Medica Veterinaria. PhD virologia pela Universidad de Buenos Aires, Argentina. Professora Titular da UFBA, docente do ICS, Departamento de Biointeração, area Microbiologia. Coordenadora do Laboratorio de Virologia 2 andar.

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Publiée

2022-05-05

Comment citer

Andrade, A. de S., Campos, G. S., Moura Costa, L. F., Monte-Alegre, A. F., & Sardi, S. I. (2022). Development of monoclonal antibodies against prM of Dengue virus 4. Revista De Ciências Médicas E Biológicas, 21(1), 40–45. https://doi.org/10.9771/cmbio.v21i1.44965

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