title: A CHO cell line engineered to express XBP1 and ERO1-Lα has increased levels of transient protein expression.
abstract: Transient gene expression (TGE) systems currently provide rapid and scalable (up to 100 L) methods for generating multigram quantities of recombinant heterologous proteins. Product titers of up to 1 g/L have been demonstrated in HEK293 cells but reported yields from Chinese hamster ovary (CHO) cells are lower at ∼300 mg/L. We report on the establishment of an engineered CHOS cell line, which has been developed for TGE. This cell line has been engineered to express both X-box binding protein (XBP-1S) and endoplasmic reticulum oxidoreductase (ERO1-Lα) and has been named CHOS-XE. CHOS-XE cells produced increased antibody (MAb) yields (5.3- 6.2 fold) in comparison to CHOS cells. Product quality was unchanged as assessed by size, charge, propensity to aggregate, major glycosylation species, and thermal stability. To further develop and test this TGE system, five commercial media were assessed, and one was shown to offer the greatest increase in antibody yields. With the addition of a commercial feed, MAb titers reached 875 mg/L.
authors: Protein Expression and Purification Group, UCB, Slough, Berkshire, SL1 4EN, England. Katharine.Cain@UCB.com
authors: Katharine
authors: K
authors: Cain
authors: Shirley
authors: S
authors: Peters
authors: Hanna
authors: H
authors: Hailu
authors: Bernie
authors: B
authors: Sweeney
authors: Paul
authors: P
authors: Stephens
authors: James
authors: J
authors: Heads
authors: Kaushik
authors: K
authors: Sarkar
authors: Andy
authors: A
authors: Ventom
authors: Catherine
authors: C
authors: Page
authors: Alan
authors: A
authors: Dickson
mesh_terms: D000818:Animals; D000911:Antibodies, Monoclonal / Q000032:analysis / Q000096:biosynthesis* / Q000378:metabolism; D016466:CHO Cells; D049109:Cell Proliferation; D006224:Cricetinae; D003412:Cricetulus; D003470:Culture Media; D004268:DNA-Binding Proteins / Q000096:biosynthesis / Q000235:genetics*; D006801:Humans; D008562:Membrane Glycoproteins / Q000096:biosynthesis / Q000235:genetics*; D010088:Oxidoreductases / Q000096:biosynthesis / Q000235:genetics*; D011994:Recombinant Proteins / Q000032:analysis / Q000096:biosynthesis / Q000378:metabolism; D000071701:Regulatory Factor X Transcription Factors; D014157:Transcription Factors / Q000096:biosynthesis / Q000235:genetics*; D000071717:X-Box Binding Protein 1
chemical_list: D000911:Antibodies, Monoclonal; D003470:Culture Media; D004268:DNA-Binding Proteins; D008562:Membrane Glycoproteins; D011994:Recombinant Proteins; D000071701:Regulatory Factor X Transcription Factors; D014157:Transcription Factors; D000071717:X-Box Binding Protein 1; C000605543:XBP1 protein, human; C405327:ERO1A protein, human; D010088:Oxidoreductases