Strategies for the Production of Soluble Interferon-Alpha Consensus and Potential Application in Arboviruses and SARS-CoV-2

Autor(es)

Grabarz, Felipe ^{[1, 2]} ; Lopes, Alexandre Paulo Yague ^[1] ; Barbosa, Flavia Ferreira ^{[2, 3]} ; Barazzone, Giovana Cappio ^[1] ; Santos, Jademilson Celestino ^[1] ; Botosso, Viviane Fongaro ^[4] ; Jorge, Soraia Attie Calil ^[5] ; Nascimento, Ana Lucia Tabet Oller ^[1] ; Astray, Renato Mancini ^[3] ; Goncalves, Viviane Maimoni ^[1]

Afiliação do(s) autor(es)

^[1] Inst Butantan, Lab Desenvolvimento Vacinas, BR-05503900 Sao Paulo - Brazil

^[2] Univ Sao Paulo, Programa Interunidades Biotecnol, BR-05508060 Sao Paulo - Brazil

^[3] Inst Butantan, Lab Multiproposito Viral, BR-05503900 Sao Paulo - Brazil

^[4] Inst Butantan, Lab Virol, BR-05503900 Sao Paulo - Brazil

^[5] Inst Butantan, Lab Biotecnol Viral, BR-05503900 Sao Paulo - Brazil

• Tipo de documento: Artigo Científico
• Fonte: LIFE-BASEL; v. 11, n. 6 JUN 2021.
• Citações Web of Science: 0

Resumo

Biopharmaceutical production is currently a multibillion-dollar industry with high growth perspectives. The research and development of biologically sourced pharmaceuticals are extremely important and a reality in our current healthcare system. Interferon alpha consensus (cIFN) is a non-natural synthetic antiviral molecule that comprises all the most prevalent amino acids of IFN-alpha into one consensus protein sequence. For clinical use, cIFN is produced in E. coli in the form of inclusion bodies. Here, we describe the use of two solubility tags (Fh8 and DsbC) to improve soluble cIFN production. Furthermore, we analyzed cIFN production in different culture media and temperatures in order to improve biopharmaceutical production. Our results demonstrate that Fh8-cIFN yield was improved when bacteria were cultivated in autoinduction culture medium at 30 degrees C. After hydrolysis, the recovery of soluble untagged cIFN was 58% from purified Fh8-cIFN molecule, fourfold higher when compared to cIFN recovered from the DsbC-cIFN, which achieved 14% recovery. The biological activity of cIFN was tested on in vitro model of antiviral effect against Zika, Mayaro, Chikungunya and SARS-CoV-2 virus infection in susceptible VERO cells. We show, for the first time, that cIFN has a potent activity against these viruses, being very low amounts of the molecule sufficient to inhibit virus multiplication. Thus, this molecule could be used in a clinical approach to treat Arboviruses and SARS-CoV-2. (AU)

• Processo FAPESP: 14/50981-0 - Busca de proteínas de superfície nas sequências do genoma da Leptospira interrogans: caracterização funcional e imunológica para o entendimento de mecanismos envolvidos na patogênese de bactéria
• Beneficiário: Ana Lucia Tabet Oller do Nascimento
• Modalidade de apoio: Auxílio à Pesquisa - Temático
• Processo FAPESP: 16/50413-8 - Administração pulmonar de uma vacina de mucosa para pneumonia
• Beneficiário: Viviane Maimoni Gonçalves
• Modalidade de apoio: Auxílio à Pesquisa - Regular