Antibiotic-Free Selection for Bio-Production: Moving Towards a New "Gold Standard"

Sodoyer, Régis; Courtois, Virginie; Peubez, Isabelle; Mignon, Charlotte

doi:10.5772/28510

Cited by 9 publications

(13 citation statements)

References 31 publications

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“…Os crescentes requerimentos regulatórios aos quais agentes biológicos estão sujeitos terão um grande impacto no campo da expressão e produção industrial de proteínas. Há uma expectativa de que, num futuro próximo, haja "tolerância zero" em relação aos sistemas de produção e seleção baseados em antibióticos (SODOYER et al, 2012).…”

Section: 52unclassified

Avaliação do sistema de estabilização plasmidial toxina-antitoxina para a produção de proteínas recombinantes em <i>Escherichia coli</i>.

Katayama¹

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Section: 52unclassified

Avaliação do sistema de estabilização plasmidial toxina-antitoxina para a produção de proteínas recombinantes em <i>Escherichia coli</i>.

Katayama¹

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“…However, there are several limitations associated with the use of E. coli expression system for the production of recombinant proteins for commercial and biomedical applications. Instability of recombinant plasmids (carrying the gene-encoding target recombinant protein) during high-density cultivation of host cells is known to result in low yield of the target recombinant proteins (Peubez et al 2010;Sodoyer et al 2012). In addition, the use of antibiotics during the cultivation of host cells (recombinant E. coli cells) expressing target recombinant proteins is a major concern (Peubez et al 2010;Sodoyer et al 2012).…”

Section: Reference Of Submitted Sequencesmentioning

confidence: 99%

Antibiotic-free expression system for the production of human interferon-beta protein

et al. 2017

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Recombinant human interferon-β (rhIFN-β), a therapeutic protein, is produced using both prokaryotic and eukaryotic expression systems. However, instability of recombinant plasmid during cultivation of results in low yield of the recombinant proteins. In addition, use of antibiotics during the cultivation imposes a major concern. In this study, we have compared the expression yield of rhIFN-β in BL21 (DE3) and SE1 cells. Gene-encoding rhIFN-β was expressed in BL21 (DE3) and SE1 cells and the cultivation of recombinant cells was done in a laboratory scale bioreactor. Our results suggest that, compared to BL21(DE3) cells, the SE1 cells expressing rhIFN-β protein can be cultivated in the medium without antibiotic and provide increased stability of recombinant plasmid and higher expression yield of rhIFN-β protein. This system can be used for the production of rhIFN-β proteins for biomedical applications.

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“…We propose to replace this resistance to an antibiotic by the Staby ® technology based on natural poison/antidote bacterial genes to insure the plasmid retention during cloning and DNA production. [10][11][12] There is a large number of poison/antidote systems, including in bacteria used industrially for production of proteins used as medical products (antigen vaccines, etc). [13][14][15][16][17] Among these systems, there is the ccd system composed of the ccdA (antidote) and ccdB genes (poison).…”

Section: Introductionmentioning

confidence: 99%

Use of Staby^®technology for development and production of DNA vaccines free of antibiotic resistance gene

Scohy

Vandermeulen

Daukandt

et al. 2013

Human Vaccines & Immunotherapeutics

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The appearance of new viruses and the cost of developing certain vaccines require that new vaccination strategies now have to be developed. DNA vaccination seems to be a particularly promising method. For this application, plasmid DNA is injected into the subject (man or animal). This plasmid DNA encodes an antigen that will be expressed by the cells of the subject. In addition to the antigen, the plasmid also encodes a resistance to an antibiotic, which is used during the construction and production steps of the plasmid. However, regulatory agencies (FDA, USDA and EMA) recommend to avoid the use of antibiotics resistance genes. Delphi Genetics developed the Staby(®) technology to replace the antibiotic-resistance gene by a selection system that relies on two bacterial genes. These genes are small in size (approximately 200 to 300 bases each) and consequently encode two small proteins. They are naturally present in the genomes of bacteria and on plasmids. The technology is already used successfully for production of recombinant proteins to achieve higher yields and without the need of antibiotics. In the field of DNA vaccines, we have now the first data validating the innocuousness of this Staby(®) technology for eukaryotic cells and the feasibility of an industrial production of an antibiotic-free DNA vaccine. Moreover, as a proof of concept, mice have been successfully vaccinated with our antibiotic-free DNA vaccine against a deadly disease, pseudorabies (induced by Suid herpesvirus-1).

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Antibiotic-Free Selection for Bio-Production: Moving Towards a New "Gold Standard"

Cited by 9 publications

References 31 publications

Avaliação do sistema de estabilização plasmidial toxina-antitoxina para a produção de proteínas recombinantes em <i>Escherichia coli</i>.

Avaliação do sistema de estabilização plasmidial toxina-antitoxina para a produção de proteínas recombinantes em <i>Escherichia coli</i>.

Antibiotic-free expression system for the production of human interferon-beta protein

Use of Staby^®technology for development and production of DNA vaccines free of antibiotic resistance gene

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Antibiotic-Free Selection for Bio-Production: Moving Towards a New "Gold Standard"

Cited by 9 publications

References 31 publications

Avaliação do sistema de estabilização plasmidial toxina-antitoxina para a produção de proteínas recombinantes em <i>Escherichia coli</i>.

Avaliação do sistema de estabilização plasmidial toxina-antitoxina para a produção de proteínas recombinantes em <i>Escherichia coli</i>.

Antibiotic-free expression system for the production of human interferon-beta protein

Use of Staby®technology for development and production of DNA vaccines free of antibiotic resistance gene

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Product

Resources

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Use of Staby^®technology for development and production of DNA vaccines free of antibiotic resistance gene