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High throughput gen...
High throughput generation of a resource of the human secretome in mammalian cells
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- Tegel, Hanna (author)
- KTH,Proteinvetenskap
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- Dannemeyer, Melanie (author)
- KTH,Proteinvetenskap
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- Kanje, Sara, 1986- (author)
- KTH,Proteinvetenskap
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show more...
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- Sivertsson, Åsa (author)
- KTH,Proteinvetenskap,Science for Life Laboratory, SciLifeLab
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- Berling, Anna (author)
- KTH,Proteinteknologi
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- Svensson, Anne-Sophie (author)
- KTH,Proteinvetenskap
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- Hober, Andreas (author)
- KTH,Proteinvetenskap,Science for Life Laboratory, SciLifeLab
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- Enstedt, Henric (author)
- KTH,Proteinteknologi
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- Volk, Anna-Luisa (author)
- KTH,Proteinteknologi
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- Lundqvist, Magnus (author)
- KTH,Proteinteknologi
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Moradi, Mona (author)
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- Afshari, Delaram (author)
- KTH,Proteinteknologi
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- Ekblad, Siri (author)
- KTH,Proteinvetenskap
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- Xu, LanLan (author)
- KTH,Proteinteknologi
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- Vestin, Malin (author)
- KTH,Proteinteknologi
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- Bidad, Faranak (author)
- KTH,Proteinteknologi
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- Schiavone, Lovisa Holmberg (author)
- AstraZeneca, R&D, Discovery Biol, Discovery Sci, Gothenburg, Sweden.
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- Davies, Rick (author)
- AstraZeneca, R&D, Discovery Biol, Discovery Sci, Cambridge, England.
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- Mayr, Lorenz M. (author)
- Syncona Investment Ltd, London, England.;Swiss Fed Inst Technol, B BIOL, Mol Hlth Sci, Zurich, Switzerland.
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- Knight, Sinead (author)
- AstraZeneca, R&D, Discovery Biol, Discovery Sci, Cambridge, England.
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- Gopel, Sven O. (author)
- AstraZeneca, BioPharmaceut R&D, Biosci Metab Res & Early Dev Cardiovasc Renal & M, Gothenburg, Sweden.
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- Voldborg, Bjorn G. (author)
- Tech Univ Denmark, Novo Nordisk Fdn Ctr Biosustainabil, Lyngby, Denmark.
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- Edfors, Fredrik (author)
- KTH,Proteinvetenskap,Science for Life Laboratory, SciLifeLab
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- Forsström, Björn (author)
- KTH,Science for Life Laboratory, SciLifeLab,Proteinteknologi
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- von Feilitzen, Kalle (author)
- KTH,Proteinteknologi,Science for Life Laboratory, SciLifeLab
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- Zwahlen, Martin (author)
- KTH,Science for Life Laboratory, SciLifeLab,Proteinteknologi
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- Rockberg, Johan (author)
- KTH,Proteinteknologi
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- Takanen, Jenny Ottosson (author)
- KTH,Proteinteknologi
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- Uhlén, Mathias (author)
- KTH,Proteinvetenskap,Science for Life Laboratory, SciLifeLab
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- Hober, Sophia, 1965- (author)
- KTH,Proteinteknologi
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(creator_code:org_t)
- Elsevier BV, 2020
- 2020
- English.
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In: New Biotechnology. - : Elsevier BV. - 1871-6784 .- 1876-4347. ; 58, s. 45-54
- Related links:
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https://doi.org/10.1...
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https://doi.org/10.1...
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https://urn.kb.se/re...
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https://doi.org/10.1...
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Abstract
Subject headings
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- The proteins secreted by human tissues and blood cells, the secretome, are important both for the basic understanding of human biology and for identification of potential targets for future diagnosis and therapy. Here, a high-throughput mammalian cell factory is presented that was established to create a resource of recombinant full-length proteins covering the majority of those annotated as 'secreted' in humans. The full-length DNA sequences of each of the predicted secreted proteins were generated by gene synthesis, the constructs were transfected into Chinese hamster ovary (CHO) cells and the recombinant proteins were produced, purified and analyzed. Almost 1,300 proteins were successfully generated and proteins predicted to be secreted into the blood were produced with a success rate of 65%, while the success rates for the other categories of secreted proteins were somewhat lower giving an overall one-pass success rate of ca. 58%. The proteins were used to generate targeted proteomics assays and several of the proteins were shown to be active in a phenotypic assay involving pancreatic beta-cell dedifferentiation. Many of the proteins that failed during production in CHO cells could be rescued in human embryonic kidney (HEK 293) cells suggesting that a cell factory of human origin can be an attractive alternative for production in mammalian cells. In conclusion, a high-throughput protein production and purification system has been successfully established to create a unique resource of the human secretome.
Subject headings
- MEDICIN OCH HÄLSOVETENSKAP -- Medicinska och farmaceutiska grundvetenskaper -- Cell- och molekylärbiologi (hsv//swe)
- MEDICAL AND HEALTH SCIENCES -- Basic Medicine -- Cell and Molecular Biology (hsv//eng)
Keyword
- secreted proteins
- protein production
- protein puri fication
- high-throughput
Publication and Content Type
- ref (subject category)
- art (subject category)
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To the university's database
- By the author/editor
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Tegel, Hanna
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Dannemeyer, Mela ...
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Kanje, Sara, 198 ...
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Sivertsson, Åsa
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Berling, Anna
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Svensson, Anne-S ...
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show more...
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Hober, Andreas
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Enstedt, Henric
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Volk, Anna-Luisa
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Lundqvist, Magnu ...
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Moradi, Mona
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Afshari, Delaram
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Ekblad, Siri
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Xu, LanLan
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Vestin, Malin
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Bidad, Faranak
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Schiavone, Lovis ...
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Davies, Rick
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Mayr, Lorenz M.
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Knight, Sinead
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Gopel, Sven O.
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Voldborg, Bjorn ...
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Edfors, Fredrik
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Forsström, Björn
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von Feilitzen, K ...
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Zwahlen, Martin
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Rockberg, Johan
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Takanen, Jenny O ...
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Uhlén, Mathias
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Hober, Sophia, 1 ...
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- About the subject
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- MEDICAL AND HEALTH SCIENCES
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MEDICAL AND HEAL ...
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and Basic Medicine
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and Cell and Molecul ...
- Articles in the publication
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New Biotechnolog ...
- By the university
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Royal Institute of Technology