Enhancing antibody folding and secretion by tailoring the Saccharomyces cerevisiae endoplasmic reticulum

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dc.contributor Aalto-yliopisto fi
dc.contributor Aalto University en
dc.contributor.author de Ruijter, Jorg C.
dc.contributor.author Koskela, Essi V.
dc.contributor.author Frey, Alexander D.
dc.date.accessioned 2016-11-17T09:38:22Z
dc.date.issued 2016-05-23
dc.identifier.citation de Ruijter , J C , Koskela , E V & Frey , A D 2016 , ' Enhancing antibody folding and secretion by tailoring the Saccharomyces cerevisiae endoplasmic reticulum ' , MICROBIAL CELL FACTORIES , vol. 15 , no. 1 , 87 . https://doi.org/10.1186/s12934-016-0488-5 en
dc.identifier.other PURE UUID: ff3dc59f-d24d-4848-bf31-9ba72f5bb319
dc.identifier.other PURE ITEMURL: https://research.aalto.fi/en/publications/enhancing-antibody-folding-and-secretion-by-tailoring-the-saccharomyces-cerevisiae-endoplasmic-reticulum(ff3dc59f-d24d-4848-bf31-9ba72f5bb319).html
dc.identifier.other PURE LINK: http://www.scopus.com/inward/record.url?scp=84969508597&partnerID=8YFLogxK
dc.identifier.other PURE FILEURL: https://research.aalto.fi/files/9199554/DeRuijter_etal_2016.pdf
dc.identifier.uri https://aaltodoc.aalto.fi/handle/123456789/23510
dc.description.abstract Background: The yeast Saccharomyces cerevisiae provides intriguing possibilities for synthetic biology and bioprocess applications, but its use is still constrained by cellular characteristics that limit the product yields. Considering the production of advanced biopharmaceuticals, a major hindrance lies in the yeast endoplasmic reticulum (ER), as it is not equipped for efficient and large scale folding of complex proteins, such as human antibodies. Results: Following the example of professional secretory cells, we show that inducing an ER expansion in yeast by deleting the lipid-regulator gene OPI1 can improve the secretion capacity of full-length antibodies up to fourfold. Based on wild-type and ER-enlarged yeast strains, we conducted a screening of a folding factor overexpression library to identify proteins and their expression levels that enhance the secretion of antibodies. Out of six genes tested, addition of the peptidyl-prolyl isomerase CPR5 provided the most beneficial effect on specific product yield while PDI1, ERO1, KAR2, LHS1 and SIL1 had a mild or even negative effect to antibody secretion efficiency. Combining genes for ER enhancement did not induce any significant additional effect compared to addition of just one element. By combining the Δopi1 strain, with the enlarged ER, with CPR5 overexpression, we were able to boost the specific antibody product yield by a factor of 10 relative to the non-engineered strain. Conclusions: Engineering protein folding in vivo is a major task for biopharmaceuticals production in yeast and needs to be optimized at several levels. By rational strain design and high-throughput screening applications we were able to increase the specific secreted antibody yields of S. cerevisiae up to 10-fold, providing a promising strain for further process optimization and platform development for antibody production. en
dc.format.mimetype application/pdf
dc.language.iso en en
dc.relation.ispartofseries MICROBIAL CELL FACTORIES en
dc.relation.ispartofseries Volume 15, issue 1 en
dc.rights openAccess en
dc.subject.other Biotechnology en
dc.subject.other Bioengineering en
dc.subject.other Applied Microbiology and Biotechnology en
dc.subject.other 220 Industrial biotechnology en
dc.title Enhancing antibody folding and secretion by tailoring the Saccharomyces cerevisiae endoplasmic reticulum en
dc.type A1 Alkuperäisartikkeli tieteellisessä aikakauslehdessä fi
dc.description.version Peer reviewed en
dc.contributor.department Department of Biotechnology and Chemical Technology
dc.contributor.department Department of Bioproducts and Biosystems en
dc.subject.keyword Antibody
dc.subject.keyword Chaperones
dc.subject.keyword Endoplasmic reticulum
dc.subject.keyword Heterologous protein production
dc.subject.keyword Protein folding
dc.subject.keyword Biotechnology
dc.subject.keyword Bioengineering
dc.subject.keyword Applied Microbiology and Biotechnology
dc.subject.keyword 220 Industrial biotechnology
dc.identifier.urn URN:NBN:fi:aalto-201611175587
dc.identifier.doi 10.1186/s12934-016-0488-5
dc.type.version publishedVersion

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