title: Improving expression of recombinant human IGF-1 using IGF-1R knockout CHO cell lines.
abstract: Chinese Hamster Ovary (CHO) cells are widely used for the large-scale production of recombinant biopharmaceuticals. However, attempts to express IGF-1 (a mutated human Insulin-like growth factor 1 Ea peptide (hIGF-1Ea mut)) in CHO cells resulted in poor cell growth and low productivity (0.1-0.2 g/L). Human IGF-1 variants negatively impacted CHO cell growth via the IGF-1 receptor (IGF-1R). Therefore knockout (KO) of the IGF-1R gene in two different CHO cell lines as well as knockdown (KD) of IGF-1R in one CHO cell line were performed. These cell line engineering approaches decreased significantly the hIGF-1 mediated cell growth inhibition and increased productivity of both KO CHO cell lines as well as of the KD CHO cell line. A productivity increase of 10-fold at pool level and sevenfold at clone level was achieved, resulting in a titer of 1.3 g/L. This data illustrate that cell line engineering approaches are powerful tools to improve the yields of recombinant proteins which are difficult to produce in CHO cells.
authors.affiliation: Integrated Biologics Profiling, Novartis Pharma, Klybeckstrasse 141, Basel, 4057, Switzerland.
authors.forename: Sandrine
authors.initials: S
authors.lastname: Romand
authors.affiliation: Integrated Biologics Profiling, Novartis Pharma, Klybeckstrasse 141, Basel, 4057, Switzerland.
authors.forename: Thomas
authors.initials: T
authors.lastname: Jostock
authors.affiliation: Musculoskeletal Diseases, Novartis Institute for BioMedical Research, Basel, Switzerland.
authors.forename: Mara
authors.initials: M
authors.lastname: Fornaro
authors.affiliation: Cell and Gene Therapies, Novartis Pharma, Basel, Switzerland.
authors.forename: Joerg
authors.initials: J
authors.lastname: Schmidt
authors.affiliation: Integrated Biologics Profiling, Novartis Pharma, Klybeckstrasse 141, Basel, 4057, Switzerland.
authors.forename: Anett
authors.initials: A
authors.lastname: Ritter
authors.affiliation: Integrated Biologics Profiling, Novartis Pharma, Klybeckstrasse 141, Basel, 4057, Switzerland.
authors.forename: Burkhard
authors.initials: B
authors.lastname: Wilms
authors.affiliation: Integrated Biologics Profiling, Novartis Pharma, Klybeckstrasse 141, Basel, 4057, Switzerland. holger.laux@novartis.com.
authors.forename: Holger
authors.initials: H
authors.lastname: Laux
mesh_terms: D000818:Animals; D016466:CHO Cells / Q000166:cytology / Q000378:metabolism*; D049109:Cell Proliferation; D006224:Cricetinae; D003412:Cricetulus; D015870:Gene Expression; D055785:Gene Knockdown Techniques / Q000379:methods; D055786:Gene Knockout Techniques; D005818:Genetic Engineering / Q000379:methods*; D006801:Humans; D007334:Insulin-Like Growth Factor I / Q000235:genetics*; D017526:Receptor, IGF Type 1 / Q000235:genetics*; D011994:Recombinant Proteins / Q000235:genetics
chemical_list: D011994:Recombinant Proteins; D007334:Insulin-Like Growth Factor I; D017526:Receptor, IGF Type 1
keywords: CHO; Chinese hamster ovary; IGF-1; IGF-1R; ZFN; cell line engineering