title: Efficient long-term and high-yielded production of a recombinant proteoglycan in eukaryotic HEK293 cells using a membrane-based bioreactor.
abstract: Standard culture systems of eukaryotic cells generally failed to deliver sufficient amounts of recombinant proteins without increasing the costs of production. We here showed that membrane-based bioreactors, initially developed for the production of monoclonal antibodies, can be very useful for the production using engineered HEK293 cells, of a recombinant proteoglycan called endocan, with achievement of high level expression and efficient long-term production. When compared to standard procedures, the growth in suspension and at high density of these cells in one bioreactor promoted a 60-fold increase of the concentration of the soluble recombinant endocan. These culture conditions did not affect cell viability, stable expression, recognition by specific monoclonal antibodies or electrophoretic profile of the recombinant endocan. Such an easy to scale up system to produce recombinant protein should open soon new opportunities to study structure and functions of endocan or any other glycosylated cell products newly investigated.
authors.affiliation: Inserm U774, Institut Pasteur de Lille, 1 rue du Professeur Calmette, 59019 Lille, France; Endotis Pharma, 59120 Loos, France.
authors.forename: Estelle
authors.initials: E
authors.lastname: Adam
authors.forename: Stéphane
authors.initials: S
authors.lastname: Sarrazin
authors.forename: Cindy
authors.initials: C
authors.lastname: Landolfi
authors.forename: Vincent
authors.initials: V
authors.lastname: Motte
authors.forename: Hugues
authors.initials: H
authors.lastname: Lortat-Jacob
authors.forename: Philippe
authors.initials: P
authors.lastname: Lassalle
authors.forename: Maryse
authors.initials: M
authors.lastname: Delehedde
mesh_terms: D019149:Bioreactors; D018929:Cell Culture Techniques / Q000379:methods*; D002460:Cell Line; D006801:Humans; D007668:Kidney / Q000502:physiology*; D008567:Membranes, Artificial; D009363:Neoplasm Proteins / Q000096:biosynthesis* / Q000235:genetics; D015202:Protein Engineering / Q000379:methods*; D011509:Proteoglycans / Q000096:biosynthesis* / Q000235:genetics; D011994:Recombinant Proteins / Q000096:biosynthesis*
chemical_list: C101134:ESM1 protein, human; D008567:Membranes, Artificial; D009363:Neoplasm Proteins; D011509:Proteoglycans; D011994:Recombinant Proteins