High throughput generation of a resource of the human secretome in mammalian cells

Hanna Tegel, Melanie Dannemeyer, Sara Kanje, Åsa Sivertsson, Anna Berling, Anne Sophie Svensson, Andreas Hober, Henric Enstedt, Anna Luisa Volk, Magnus Lundqvist, Mona Moradi, Delaram Afshari, Siri Ekblad, Lan Lan Xu, Malin Westin, Faranak Bidad, Lovisa Holmberg Schiavone, Rick Davies, Lorenz M. Mayr, Sinead KnightSven O. Göpel, Björn G. Voldborg, Fredrik Edfors, Björn Forsström, Kalle von Feilitzen, Martin Zwahlen, Johan Rockberg, Jenny Ottosson Takanen, Mathias Uhlén, Sophia Hober*

*Corresponding author for this work

Research output: Contribution to journalJournal articleResearchpeer-review

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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 β-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.
Original languageEnglish
JournalNew Biotechnology
Pages (from-to)45-54
Number of pages10
Publication statusPublished - 2020


  • Secreted proteins
  • Protein production
  • Protein purification
  • High-throughput


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