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Fast and robust recombinant protein production utilizing episomal stable pools in WAVE bioreactors
KTH, School of Engineering Sciences in Chemistry, Biotechnology and Health (CBH), Protein Science, Protein Technology.ORCID iD: 0000-0001-9542-601x
KTH, School of Engineering Sciences in Chemistry, Biotechnology and Health (CBH), Protein Science, Protein Technology.ORCID iD: 0009-0000-3517-8042
KTH, School of Engineering Sciences in Chemistry, Biotechnology and Health (CBH), Protein Science, Protein Technology.ORCID iD: 0000-0002-4751-2519
KTH, School of Engineering Sciences in Chemistry, Biotechnology and Health (CBH), Protein Science, Protein Technology.ORCID iD: 0000-0002-4033-1205
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2024 (English)In: Protein Expression and Purification, ISSN 1046-5928, E-ISSN 1096-0279, Vol. 221, article id 106505Article in journal (Refereed) Published
Abstract [en]

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.

Place, publisher, year, edition, pages
Elsevier BV , 2024. Vol. 221, article id 106505
Keywords [en]
Drug discovery, Early development, Episomal stable pools, Protein expression, Recombinant proteins
National Category
Biological Sciences
Identifiers
URN: urn:nbn:se:kth:diva-347279DOI: 10.1016/j.pep.2024.106505ISI: 001247292800001Scopus ID: 2-s2.0-85194428753OAI: oai:DiVA.org:kth-347279DiVA, id: diva2:1867211
Note

QC 20240702

Available from: 2024-06-10 Created: 2024-06-10 Last updated: 2024-07-02Bibliographically approved

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Dannemeyer, MelanieBerling, AnnaKanje, SaraEnstedt, HenricAfshari, DelaramVestin, MalinJensen, GabriellaUhlén, MathiasHober, SophiaTegel, Hanna

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Dannemeyer, MelanieBerling, AnnaKanje, SaraEnstedt, HenricXu, LanLanAfshari, DelaramVestin, MalinJensen, GabriellaUhlén, MathiasHober, SophiaTegel, Hanna
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