Plasmid Replicons for the Production of Pharmaceutical-Grade pDNA, Proteins and Antigens by Lactococcus lactis Cell Factories

Duarte, Sofia O. D.; Monteiro, Gabriel A.

doi:10.3390/ijms22031379

Cited by 7 publications

(6 citation statements)

References 194 publications

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“…Moreover, mice immunized with double-layered protein nanoparticles containing M2e and HA were protected from challenge with H1N1, H3N2, H5N1, and H7N9 viruses [ 19 ]. Further, an M1-HA2 fusion protein expressed by recombinant Lactococcus lactis provided mucosal protection against the H9N2 virus in chickens [ 139 ].…”

Section: Multiple Proteins/peptides-based Uivsmentioning

confidence: 99%

Targeting Antigens for Universal Influenza Vaccine Development

Nguyen

Choi

2021

Viruses

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Traditional influenza vaccines generate strain-specific antibodies which cannot provide protection against divergent influenza virus strains. Further, due to frequent antigenic shifts and drift of influenza viruses, annual reformulation and revaccination are required in order to match circulating strains. Thus, the development of a universal influenza vaccine (UIV) is critical for long-term protection against all seasonal influenza virus strains, as well as to provide protection against a potential pandemic virus. One of the most important strategies in the development of UIVs is the selection of optimal targeting antigens to generate broadly cross-reactive neutralizing antibodies or cross-reactive T cell responses against divergent influenza virus strains. However, each type of target antigen for UIVs has advantages and limitations for the generation of sufficient immune responses against divergent influenza viruses. Herein, we review current strategies and perspectives regarding the use of antigens, including hemagglutinin, neuraminidase, matrix proteins, and internal proteins, for universal influenza vaccine development.

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Section: Multiple Proteins/peptides-based Uivsmentioning

confidence: 99%

Targeting Antigens for Universal Influenza Vaccine Development

Nguyen

Choi

2021

Viruses

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“…Lactococcus lactis . In contrary to E. coli, L. lactis is generally regarded as safe (GRAS) and thus simplifies the downstream processing [26, 27].…”

Section: Introductionmentioning

confidence: 99%

Sulfate limitation increases specific plasmid DNA yield and productivity inE. colifed-batch processes

Gotsmy

Stobl

Weiß

et al. 2023

Preprint

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Plasmid DNA (pDNA) is a key biotechnological product whose importance became apparent in the last years due to its role as a raw material in the messenger ribonucleic acid (mRNA) vaccine manufacturing process. In pharmaceutical production processes, cells need to grow in the defined medium in order to guarantee the highest standards of quality and repeatability. However, often these requirements result in low product titer, productivity, and yield. In this study, we used constraint-based metabolic modeling to optimize the average volumetric productivity of pDNA production in a fed-batch process. We identified a set of 13 nutrients in the growth medium that are essential for cell growth but not for pDNA replication. When these nutrients are depleted in the medium, cell growth is stalled and pDNA production is increased, raising the specific and volumetric yield and productivity. To exploit this effect we designed a three-stage process (1. batch, 2. fed-batch with cell growth, 3. fed-batch without cell growth). The transition between stage 2 and 3 is induced by sulfate starvation. Its onset can be easily controlled via the initial concentration of sulfate in the medium. We validated the decoupling behavior of sulfate and assessed pDNA quality attributes (supercoiled pDNA content) in E. coli with lab-scale bioreactor cultivations. The results showed an increase in supercoiled pDNA to biomass yield by 29% upon limitation of sulfate. In conclusion, even for routinely manufactured biotechnological products such as pDNA, simple changes in the growth medium can significantly improve the yield and quality.

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“…Lactococcus lactis . Contrary to E. coli , L. lactis is generally regarded as safe (GRAS) and thus simplifies the downstream processing [ 27 , 28 ].…”

Section: Introductionmentioning

confidence: 99%

Sulfate limitation increases specific plasmid DNA yield and productivity in E. coli fed-batch processes

Gotsmy,

Strobl,

Weiß

et al. 2023

Microb Cell Fact

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Plasmid DNA (pDNA) is a key biotechnological product whose importance became apparent in the last years due to its role as a raw material in the messenger ribonucleic acid (mRNA) vaccine manufacturing process. In pharmaceutical production processes, cells need to grow in the defined medium in order to guarantee the highest standards of quality and repeatability. However, often these requirements result in low product titer, productivity, and yield. In this study, we used constraint-based metabolic modeling to optimize the average volumetric productivity of pDNA production in a fed-batch process. We identified a set of 13 nutrients in the growth medium that are essential for cell growth but not for pDNA replication. When these nutrients are depleted in the medium, cell growth is stalled and pDNA production is increased, raising the specific and volumetric yield and productivity. To exploit this effect we designed a three-stage process (1. batch, 2. fed-batch with cell growth, 3. fed-batch without cell growth). The transition between stage 2 and 3 is induced by sulfate starvation. Its onset can be easily controlled via the initial concentration of sulfate in the medium. We validated the decoupling behavior of sulfate and assessed pDNA quality attributes (supercoiled pDNA content) in E. coli with lab-scale bioreactor cultivations. The results showed an increase in supercoiled pDNA to biomass yield by 33% and an increase of supercoiled pDNA volumetric productivity by 13 % upon limitation of sulfate. In conclusion, even for routinely manufactured biotechnological products such as pDNA, simple changes in the growth medium can significantly improve the yield and quality. Graphical Abstract

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Plasmid Replicons for the Production of Pharmaceutical-Grade pDNA, Proteins and Antigens by Lactococcus lactis Cell Factories

Cited by 7 publications

References 194 publications

Targeting Antigens for Universal Influenza Vaccine Development

Targeting Antigens for Universal Influenza Vaccine Development

Sulfate limitation increases specific plasmid DNA yield and productivity inE. colifed-batch processes

Sulfate limitation increases specific plasmid DNA yield and productivity in E. coli fed-batch processes

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