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Fast and robust recombinant protein production utilizing episomal stable pools in WAVE bioreactors

Dannemeyer, Melanie (author)
KTH,Proteinteknologi
Berling, Anna (author)
KTH,Proteinteknologi
Kanje, Sara, 1986- (author)
KTH,Proteinteknologi
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Enstedt, Henric (author)
KTH,Proteinteknologi
Xu, LanLan (author)
KTH,Proteinteknologi
Afshari, Delaram (author)
KTH,Proteinteknologi
Vestin, Malin (author)
KTH,Proteinteknologi
Jensen, Gabriella (author)
KTH,Proteinvetenskap
Uhlén, Mathias (author)
KTH,Systembiologi
Hober, Sophia, Professor, 1965- (author)
KTH,Proteinteknologi
Tegel, Hanna (author)
KTH,Proteinteknologi
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 (creator_code:org_t)
Elsevier BV, 2024
2024
English.
In: Protein Expression and Purification. - : Elsevier BV. - 1046-5928 .- 1096-0279. ; 221
  • Journal article (peer-reviewed)
Abstract Subject headings
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  • Protein reagents are essential resources for several stages of drug discovery projects from structural biology and assay development through lead optimization. Depending on the aim of the project different amounts of pure protein are required. Small-scale expressions are initially used to determine the reachable levels of production and quality before scaling up protein reagent supply. Commonly, amounts of several hundreds of milligrams to grams are needed for different experiments, including structural investigations and activity evaluations, which require rather large cultivation volumes. This implies that cultivation of large volumes of either transiently transfected cells or stable pools/stable cell lines is needed. Hence, a production process that is scalable, speeds up the development projects, and increases the robustness of protein reagent quality throughout scales. Here we present a protein production pipeline with high scalability. We show that our protocols for protein production in Chinese hamster ovary cells allow for a seamless and efficient scale-up with robust product quality and high performance. The flexible scale of the production process, as shown here, allows for shorter lead times in drug discovery projects where there is a reagent demand for a specific protein or a set of target proteins.

Subject headings

NATURVETENSKAP  -- Biologi (hsv//swe)
NATURAL SCIENCES  -- Biological Sciences (hsv//eng)

Keyword

Drug discovery
Early development
Episomal stable pools
Protein expression
Recombinant proteins

Publication and Content Type

ref (subject category)
art (subject category)

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