A CHO cell line engineered to express XBP1 and ERO1‐Lα has increased levels of transient protein expression
- 19 January 2013
- journal article
- research article
- Published by Wiley in Biotechnology Progress
- Vol. 29 (3), 697-706
- https://doi.org/10.1002/btpr.1693
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 1 but reported yields from Chinese hamster ovary (CHO) cells are lower at ∼300 mg/L. 2 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. © 2013 American Institute of Chemical Engineers Biotechnol. Prog., 29:697–706, 2013Keywords
This publication has 29 references indexed in Scilit:
- Identifying bottlenecks in transient and stable production of recombinant monoclonal‐antibody sequence variants in chinese hamster ovary cellsBiotechnology Progress, 2012
- A simple high-yielding process for transient gene expression in CHO cellsJournal of Biotechnology, 2011
- Effect of inducible co‐overexpression of protein disulfide isomerase and endoplasmic reticulum oxidoreductase on the specific antibody productivity of recombinant Chinese hamster ovary cellsBiotechnology & Bioengineering, 2010
- Valproic acid enhances recombinant mRNA and protein levels in transiently transfected Chinese hamster ovary cellsJournal of Biotechnology, 2010
- Evaluation of a combinatorial cell engineering approach to overcome apoptotic effects in XBP-1(s) expressing cellsJournal of Biotechnology, 2010
- Regulation of XBP‐1 signaling during transient and stable recombinant protein production in CHO cellsBiotechnology Progress, 2009
- Coexpression of acidic fibroblast growth factor enhances specific productivity and antibody titers in transiently transfected HEK293 cellsNew Biotechnology, 2008
- Antioxidants reduce endoplasmic reticulum stress and improve protein secretionProceedings of the National Academy of Sciences of the United States of America, 2008
- Recombinant protein production by large-scale transient gene expression in mammalian cells: state of the art and future perspectivesBiotechnology Letters, 2007
- Production of recombinant protein therapeutics in cultivated mammalian cellsNature Biotechnology, 2004