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Fast and robust recombinant protein production utilizing episomal stable pools in WAVE bioreactors.
Dannemeyer, Melanie; Berling, Anna; Kanje, Sara; Enstedt, Henric; Xu, LanLan; Afshari, Delaram; Westin, Malin; Hober, Gabriella; Uhlén, Mathias; Hober, Sophia; Tegel, Hanna.
Afiliação
  • Dannemeyer M; Department of Protein Science, School of Chemistry, Biotechnology and Health, KTH - Royal Institute of Technology, SE-10691, Stockholm, Sweden.
  • Berling A; Department of Protein Science, School of Chemistry, Biotechnology and Health, KTH - Royal Institute of Technology, SE-10691, Stockholm, Sweden.
  • Kanje S; Department of Protein Science, School of Chemistry, Biotechnology and Health, KTH - Royal Institute of Technology, SE-10691, Stockholm, Sweden.
  • Enstedt H; Department of Protein Science, School of Chemistry, Biotechnology and Health, KTH - Royal Institute of Technology, SE-10691, Stockholm, Sweden.
  • Xu L; Department of Protein Science, School of Chemistry, Biotechnology and Health, KTH - Royal Institute of Technology, SE-10691, Stockholm, Sweden.
  • Afshari D; Department of Protein Science, School of Chemistry, Biotechnology and Health, KTH - Royal Institute of Technology, SE-10691, Stockholm, Sweden.
  • Westin M; Department of Protein Science, School of Chemistry, Biotechnology and Health, KTH - Royal Institute of Technology, SE-10691, Stockholm, Sweden.
  • Hober G; Department of Protein Science, School of Chemistry, Biotechnology and Health, KTH - Royal Institute of Technology, SE-10691, Stockholm, Sweden.
  • Uhlén M; Department of Protein Science, School of Chemistry, Biotechnology and Health, KTH - Royal Institute of Technology, SE-10691, Stockholm, Sweden.
  • Hober S; Department of Protein Science, School of Chemistry, Biotechnology and Health, KTH - Royal Institute of Technology, SE-10691, Stockholm, Sweden.
  • Tegel H; Department of Protein Science, School of Chemistry, Biotechnology and Health, KTH - Royal Institute of Technology, SE-10691, Stockholm, Sweden. Electronic address: hannat@kth.se.
Protein Expr Purif ; 221: 106505, 2024 Sep.
Article em En | MEDLINE | ID: mdl-38768672
ABSTRACT
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.
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Texto completo: 1 Coleções: 01-internacional Base de dados: MEDLINE Assunto principal: Plasmídeos / Proteínas Recombinantes / Cricetulus / Reatores Biológicos Limite: Animals Idioma: En Revista: Protein Expr Purif Ano de publicação: 2024 Tipo de documento: Article

Texto completo: 1 Coleções: 01-internacional Base de dados: MEDLINE Assunto principal: Plasmídeos / Proteínas Recombinantes / Cricetulus / Reatores Biológicos Limite: Animals Idioma: En Revista: Protein Expr Purif Ano de publicação: 2024 Tipo de documento: Article