دورية أكاديمية
Sf9 Cell Metabolism Throughout the Recombinant Baculovirus and Rabies Virus-Like Particles Production in Two Culture Systems.
العنوان: | Sf9 Cell Metabolism Throughout the Recombinant Baculovirus and Rabies Virus-Like Particles Production in Two Culture Systems. |
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المؤلفون: | Guardalini LGO; Laboratório de Biotecnologia Viral, Instituto Butantan, Av. Vital Brasil 1500, São Paulo, SP, CEP 05503-900, Brazil., Leme J; Laboratório de Biotecnologia Viral, Instituto Butantan, Av. Vital Brasil 1500, São Paulo, SP, CEP 05503-900, Brazil., da Silva Cavalcante PE; Laboratório de Biotecnologia Viral, Instituto Butantan, Av. Vital Brasil 1500, São Paulo, SP, CEP 05503-900, Brazil., de Mello RG; Laboratório de Biotecnologia Viral, Instituto Butantan, Av. Vital Brasil 1500, São Paulo, SP, CEP 05503-900, Brazil., Bernardino TC; Laboratório de Biotecnologia Viral, Instituto Butantan, Av. Vital Brasil 1500, São Paulo, SP, CEP 05503-900, Brazil., Jared SGS; Laboratório de Biologia Estrutural, Instituto Butantan, Av. Vital Brasil 1500, São Paulo, SP, CEP 05503-900, Brazil., Antoniazzi MM; Laboratório de Biologia Estrutural, Instituto Butantan, Av. Vital Brasil 1500, São Paulo, SP, CEP 05503-900, Brazil., Astray RM; Laboratório Multipropósito, Instituto Butantan, Av. Vital Brasil 1500, São Paulo, SP, CEP 05503-900, Brazil., Tonso A; Laboratório de Células Animais, Departamento de Engenharia Química, Escola Politécnica, Universidade de São Paulo, Av. Prof. Luciano Gualberto, Trav. 3, 380, São Paulo, SP, 05508-900, Brazil., Núñez EGF; Grupo de Engenharia de Bioprocessos. Escola de Artes, Ciências e Humanidades (EACH), Universidade de São Paulo, Rua Arlindo Béttio, 1000, São Paulo, SP, CEP 03828-000, Brazil., Jorge SAC; Laboratório de Biotecnologia Viral, Instituto Butantan, Av. Vital Brasil 1500, São Paulo, SP, CEP 05503-900, Brazil. soraia.jorge@butantan.gov.br. |
المصدر: | Molecular biotechnology [Mol Biotechnol] 2024 Feb; Vol. 66 (2), pp. 354-364. Date of Electronic Publication: 2023 May 10. |
نوع المنشور: | Journal Article |
اللغة: | English |
بيانات الدورية: | Publisher: Springer Country of Publication: Switzerland NLM ID: 9423533 Publication Model: Print-Electronic Cited Medium: Internet ISSN: 1559-0305 (Electronic) Linking ISSN: 10736085 NLM ISO Abbreviation: Mol Biotechnol Subsets: MEDLINE |
أسماء مطبوعة: | Publication: [Cham] : Springer Original Publication: Totowa, NJ : Humana Press, c1994- |
مواضيع طبية MeSH: | Rabies virus*/genetics , Rabies* , Ammonium Compounds*, Animals ; Sf9 Cells ; Glutamine ; Baculoviridae/genetics ; Recombinant Proteins/genetics ; Culture Media, Serum-Free ; Glutamic Acid ; Lactates ; Glucose ; Spodoptera |
مستخلص: | This work aimed to assess the Sf9 cell metabolism during growth, and infection steps with recombinant baculovirus bearing rabies virus proteins, to finally obtain rabies VLP in two culture systems: Schott flask (SF) and stirred tank reactor (STR). Eight assays were performed in SF and STR (four assays in each system) using serum-free SF900 III culture medium. Two non-infection growth kinetics assays and six recombinant baculovirus infection assays. The infection runs were carried out at 0.1 pfu/cell multiplicity of infection (MOI) for single baculovirus bearing rabies glycoprotein (BVG) and matrix protein (BVM) and a coinfection with both baculoviruses at MOI of 3 and 2 pfu/cell for BVG and BVM, respectively. The SF assays were done in triplicate. The glucose, glutamine, glutamate, lactate, and ammonium uptake or release specific rates were quantified over the exponential growth phase and infection stage. The highest uptake specific rate was observed for glucose (42.5 × 10 -12 mmol cell/h) in SF and for glutamine (30.8 × 10 -12 mmol/cell/h) in STR, in the exponential growth phases. A wave pattern was observed for assessed analytes throughout the infection phase and the glucose had the highest wave amplitude within the 10 -10 mmol cell/h order. This alternative uptake and release behavior is in harmony with the lytic cycle of baculovirus in insect cells. The virus propagation and VLP generation were not limited by glucose, glutamine, and glutamate, neither by the toxicity of lactate nor ammonium under the conditions appraised in this work. The findings from this work can be useful to set baculovirus infection processes at high cell density to improve rabies VLP yield, purity, and productivity. (© 2023. The Author(s), under exclusive licence to Springer Science+Business Media, LLC, part of Springer Nature.) |
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معلومات مُعتمدة: | 2018/10538-1 Fundação de Amparo à Pesquisa do Estado de São Paulo; 2016/22780-6 Fundação de Amparo à Pesquisa do Estado de São Paulo; 168539/2018-7 Conselho Nacional de Desenvolvimento Científico e Tecnológico |
فهرسة مساهمة: | Keywords: Insect cell metabolism; Rabies virus-like particles; Recombinant baculovirus; Sf9 cells; Stirred tank bioreactor; Viral infection |
المشرفين على المادة: | 0RH81L854J (Glutamine) 0 (Recombinant Proteins) 0 (Culture Media, Serum-Free) 3KX376GY7L (Glutamic Acid) 0 (Lactates) IY9XDZ35W2 (Glucose) 0 (Ammonium Compounds) |
تواريخ الأحداث: | Date Created: 20230510 Date Completed: 20240124 Latest Revision: 20240124 |
رمز التحديث: | 20240124 |
DOI: | 10.1007/s12033-023-00759-2 |
PMID: | 37162721 |
قاعدة البيانات: | MEDLINE |
تدمد: | 1559-0305 |
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DOI: | 10.1007/s12033-023-00759-2 |