Maximization of the Minicircle DNA Vaccine Production Expressing SARS-CoV-2 RBD
| Autor(a) principal: | |
|---|---|
| Data de Publicação: | 2022 |
| Outros Autores: | , , , , , , |
| Tipo de documento: | Artigo |
| Idioma: | eng |
| Título da fonte: | Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos) |
| Texto Completo: | http://hdl.handle.net/10362/143759 |
Resumo: | LA/P/0140/2020 of the Associate Laboratory Institute for Health and Bioeconomy-i4HB. Diana Costa acknowledges research program contract I(SFRH/BD/10201/2020. Publisher Copyright: © 2022 by the authors. Licensee MDPI, Basel, Switzerland. |
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Maximization of the Minicircle DNA Vaccine Production Expressing SARS-CoV-2 RBDbioreactorCOVID-19design of experimentsminicircle DNA vaccineMedicine (miscellaneous)Biochemistry, Genetics and Molecular Biology(all)SDG 3 - Good Health and Well-beingLA/P/0140/2020 of the Associate Laboratory Institute for Health and Bioeconomy-i4HB. Diana Costa acknowledges research program contract I(SFRH/BD/10201/2020. Publisher Copyright: © 2022 by the authors. Licensee MDPI, Basel, Switzerland.Nucleic acid vaccines have been proven to be a revolutionary technology to induce an efficient, safe and rapid response against pandemics, like the coronavirus disease (COVID-19). Minicircle DNA (mcDNA) is an innovative vector more stable than messenger RNA and more efficient in cell transfection and transgene expression than conventional plasmid DNA. This work describes the construction of a parental plasmid (PP) vector encoding the receptor-binding domain (RBD) of the S protein from severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), and the use of the Design of Experiments (DoE) to optimize PP recombination into mcDNA vector in an orbital shaker. First, the results revealed that host cells should be grown at 42◦C and the Terrific Broth (TB) medium should be replaced by Luria Broth (LB) medium containing 0.01% L-arabinose for the induction step. The antibiotic concentration, the induction time, and the induction temperature were used as DoE inputs to maximize the % of recombined mcDNA. The quadratic model was statistically significant (p-value < 0.05) and presented a non-significant lack of fit (p-value > 0.05) with a suitable coefficient of determination. The optimal point was validated using 1 h of induction, at 30◦C, without the presence of antibiotics, obtaining 93.87% of recombined mcDNA. Based on these conditions, the production of mcDNA was then maximized in a mini-bioreactor platform. The most favorable condition obtained in the bioreactor was obtained by applying 60% pO2 in the fermentation step during 5 h and 30% pO2 in the induction step, with 0.01% L-arabinose throughout 5 h. The yield of mcDNA-RBD was increased to a concentration of 1.15 g/L, when compared to the orbital shaker studies (16.48 mg/L). These data revealed that the bioreactor application strongly incremented the host biomass yield and simultaneously improved the recombination levels of PP into mcDNA. Altogether, these results contributed to improving mcDNA-RBD biosynthesis to make the scale-up of mcDNA manufacture simpler, cost-effective, and attractive for the biotechnology industry.UCIBIO - Applied Molecular Biosciences UnitDQ - Departamento de QuímicaRUNVentura, CathyEusébio, DalindaGonçalves, Ana M.Barroca-Ferreira, JorgeCosta, DianaCui, ZhengrongPassarinha, Luís A.Sousa, Ângela2022-09-15T22:42:05Z2022-04-252022-04-25T00:00:00Zinfo:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/article18application/pdfhttp://hdl.handle.net/10362/143759eng2227-9059PURE: 46298779https://doi.org/10.3390/biomedicines10050990info:eu-repo/semantics/openAccessreponame:Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos)instname:Agência para a Sociedade do Conhecimento (UMIC) - FCT - Sociedade da Informaçãoinstacron:RCAAP2024-03-11T05:22:28Zoai:run.unl.pt:10362/143759Portal AgregadorONGhttps://www.rcaap.pt/oai/openaireopendoar:71602024-03-20T03:51:07.669626Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos) - Agência para a Sociedade do Conhecimento (UMIC) - FCT - Sociedade da Informaçãofalse |
| dc.title.none.fl_str_mv |
Maximization of the Minicircle DNA Vaccine Production Expressing SARS-CoV-2 RBD |
| title |
Maximization of the Minicircle DNA Vaccine Production Expressing SARS-CoV-2 RBD |
| spellingShingle |
Maximization of the Minicircle DNA Vaccine Production Expressing SARS-CoV-2 RBD Ventura, Cathy bioreactor COVID-19 design of experiments minicircle DNA vaccine Medicine (miscellaneous) Biochemistry, Genetics and Molecular Biology(all) SDG 3 - Good Health and Well-being |
| title_short |
Maximization of the Minicircle DNA Vaccine Production Expressing SARS-CoV-2 RBD |
| title_full |
Maximization of the Minicircle DNA Vaccine Production Expressing SARS-CoV-2 RBD |
| title_fullStr |
Maximization of the Minicircle DNA Vaccine Production Expressing SARS-CoV-2 RBD |
| title_full_unstemmed |
Maximization of the Minicircle DNA Vaccine Production Expressing SARS-CoV-2 RBD |
| title_sort |
Maximization of the Minicircle DNA Vaccine Production Expressing SARS-CoV-2 RBD |
| author |
Ventura, Cathy |
| author_facet |
Ventura, Cathy Eusébio, Dalinda Gonçalves, Ana M. Barroca-Ferreira, Jorge Costa, Diana Cui, Zhengrong Passarinha, Luís A. Sousa, Ângela |
| author_role |
author |
| author2 |
Eusébio, Dalinda Gonçalves, Ana M. Barroca-Ferreira, Jorge Costa, Diana Cui, Zhengrong Passarinha, Luís A. Sousa, Ângela |
| author2_role |
author author author author author author author |
| dc.contributor.none.fl_str_mv |
UCIBIO - Applied Molecular Biosciences Unit DQ - Departamento de Química RUN |
| dc.contributor.author.fl_str_mv |
Ventura, Cathy Eusébio, Dalinda Gonçalves, Ana M. Barroca-Ferreira, Jorge Costa, Diana Cui, Zhengrong Passarinha, Luís A. Sousa, Ângela |
| dc.subject.por.fl_str_mv |
bioreactor COVID-19 design of experiments minicircle DNA vaccine Medicine (miscellaneous) Biochemistry, Genetics and Molecular Biology(all) SDG 3 - Good Health and Well-being |
| topic |
bioreactor COVID-19 design of experiments minicircle DNA vaccine Medicine (miscellaneous) Biochemistry, Genetics and Molecular Biology(all) SDG 3 - Good Health and Well-being |
| description |
LA/P/0140/2020 of the Associate Laboratory Institute for Health and Bioeconomy-i4HB. Diana Costa acknowledges research program contract I(SFRH/BD/10201/2020. Publisher Copyright: © 2022 by the authors. Licensee MDPI, Basel, Switzerland. |
| publishDate |
2022 |
| dc.date.none.fl_str_mv |
2022-09-15T22:42:05Z 2022-04-25 2022-04-25T00:00:00Z |
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info:eu-repo/semantics/publishedVersion |
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info:eu-repo/semantics/article |
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article |
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publishedVersion |
| dc.identifier.uri.fl_str_mv |
http://hdl.handle.net/10362/143759 |
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http://hdl.handle.net/10362/143759 |
| dc.language.iso.fl_str_mv |
eng |
| language |
eng |
| dc.relation.none.fl_str_mv |
2227-9059 PURE: 46298779 https://doi.org/10.3390/biomedicines10050990 |
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info:eu-repo/semantics/openAccess |
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openAccess |
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18 application/pdf |
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reponame:Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos) instname:Agência para a Sociedade do Conhecimento (UMIC) - FCT - Sociedade da Informação instacron:RCAAP |
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Agência para a Sociedade do Conhecimento (UMIC) - FCT - Sociedade da Informação |
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RCAAP |
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RCAAP |
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Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos) |
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Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos) |
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Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos) - Agência para a Sociedade do Conhecimento (UMIC) - FCT - Sociedade da Informação |
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