Cellular targets for improved manufacturing of virus-based biopharmaceuticals in animal cells

Rodrigues, Ana Paula Leite; Carrondo, Manuel J.T.; Alves, Paula M.; Coroadinha, Ana S.

doi:10.1016/j.tibtech.2014.09.010

Cited by 23 publications

(21 citation statements)

References 61 publications

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“…Likewise, the production of complex enveloped VLPs will profit from the establishment of stable cell lines where a continuous production is enabled, facilitating bioreaction and purification processes (avoiding the need to produce plasmid DNA or baculovirus stocks and their subsequent removal from the final product). On the other hand, to increase the production yields and reduce the costs cell engineering strategies may be envisaged [95]. For simplicity purposes, we shall distinguish cell line development from cell line engineering, as illustrated in Fig.…”

Section: Cell Line Development and Engineering: Current And Future Trmentioning

confidence: 99%

Viral vaccines and their manufacturing cell substrates: New trends and designs in modern vaccinology

Rodrigues

Soares

Guerreiro

et al. 2015

Biotechnology Journal

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Vaccination is one of the most effective interventions in global health. The worldwide vaccination programs significantly reduced the number of deaths caused by infectious agents. A successful example was the eradication of smallpox in 1979 after two centuries of vaccination campaigns. Since the first variolation administrations until today, the knowledge on immunology has increased substantially. This knowledge combined with the introduction of cell culture and DNA recombinant technologies revolutionized vaccine design. This review will focus on vaccines against human viral pathogens, recent developments on vaccine design and cell substrates used for their manufacture. While the production of attenuated and inactivated vaccines requires the use of the respective permissible cell substrates, the production of recombinant antigens, virus-like particles, vectored vaccines and chimeric vaccines requires the use - and often the development - of specific cell lines. Indeed, the development of novel modern viral vaccine designs combined with, the stringent safety requirements for manufacture, and the better understanding on animal cell metabolism and physiology are increasing the awareness on the importance of cell line development and engineering areas. A new era of modern vaccinology is arriving, offering an extensive toolbox to materialize novel and creative ideas in vaccine design and its manufacture.

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Section: Cell Line Development and Engineering: Current And Future Trmentioning

confidence: 99%

Viral vaccines and their manufacturing cell substrates: New trends and designs in modern vaccinology

Rodrigues

Soares

Guerreiro

et al. 2015

Biotechnology Journal

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“…Metabolomics analyses of several cell lines have shown that viral infection activates large number of metabolic processes [8,[25][26][27][28] unlike the metabolic reduction observed in the case of recombinant protein production [29]. Infection induced metabolic shift has been extensively studied and confirmed for diverse cell/virus production platforms (rubella virus [30][31][32], cytomegalovirus [25,33], mayaro virus [34], dengue virus [35], mumps virus [36], newcastle-disease virus [36], polio virus [37][38][39], reovirus [40], influenza viruses [41]).…”

Section: Metabolomics To Improve Viral Productions In Hek293 Cell Culmentioning

confidence: 99%

“…Viral replication induces major changes in the cell physiology and metabolic states [8]. The cell metabolic efficiency, i.e.…”

Section: Introductionmentioning

confidence: 99%

Influence of HEK293 metabolism on the production of viral vectors and vaccine

Petiot

Čuperlović-Culf

Shen

et al. 2015

Vaccine

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“…In 293 FLEX cell line, the transcription of both the viral transgene and the Gag-pol are under the control of the retroviral long terminal repeat (LTR). Herein, an improved metabolic status refers to a conjugation of active pathways potentially favoring virus production (Rodrigues et al, 2014). In fact, the retroviral LTR has been previously suggested to exhibit a transcriptionally dynamic behavior, with more than 50-fold increase in the LTR-driven mRNA, in response to changes in the culture medium (Olsen and Sechelski, 1995).…”

Section: Discussionmentioning

confidence: 99%

Increased titer and reduced lactate accumulation in recombinant retrovirus production through the down‐regulation of HIF1 and PDK

Rodrigues

Guerreiro

Formas-Oliveira

et al. 2015

Biotech & Bioengineering

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Many mammalian cell lines used in the manufacturing of biopharmaceuticals exhibit high glycolytic flux predominantly channeled to the production of lactate. The accumulation of lactate in culture reduces cell viability and may also decrease product quality. In this work, we engineered a HEK 293 derived cell line producing a recombinant gene therapy retroviral vector, by down-regulating hypoxia inducible factor 1 (HIF1) and pyruvate dehydrogenase kinase (PDK). Specific productivity of infectious viral titers could be increased more than 20-fold for single gene knock-down (HIF1 or PDK) and more than 30-fold under combined down-regulation. Lactate production was reduced up to 4-fold. However, the reduction in lactate production, alone, was not sufficient to enhance the titer: high-titer clones also showed significant enrollment of metabolic routes not related to lactate production. Transcriptome analysis indicated activation of biological amines metabolism, detoxification routes, including glutathione metabolism, pentose phosphate pathway, glycogen biosynthesis and amino acid catabolism. The latter were validated by enzyme activity assays and metabolite profiling, respectively. High-titer clones also presented substantially increased transcript levels of the viral genes expression cassettes. The results herein presented demonstrate the impact of HIF1 and PDK down-regulation on the production performance of a mammalian cell line, reporting one of the highest fold-increase in specific productivity of infectious virus titers achieved by metabolic engineering. They additionally highlight the contribution of secondary pathways, beyond those related to lactate production, that can be also explored to pursue improved metabolic status favoring a high-producing phenotype.

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Cellular targets for improved manufacturing of virus-based biopharmaceuticals in animal cells

Cited by 23 publications

References 61 publications

Viral vaccines and their manufacturing cell substrates: New trends and designs in modern vaccinology

Viral vaccines and their manufacturing cell substrates: New trends and designs in modern vaccinology

Influence of HEK293 metabolism on the production of viral vectors and vaccine

Increased titer and reduced lactate accumulation in recombinant retrovirus production through the down‐regulation of HIF1 and PDK

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