CHO cell engineering to prevent polypeptide aggregation and improve therapeutic protein secretion.

Details

Serval ID
serval:BIB_C2E58E538DD5
Type
Article: article from journal or magazin.
Collection
Publications
Title
CHO cell engineering to prevent polypeptide aggregation and improve therapeutic protein secretion.
Journal
Metabolic Engineering
Author(s)
Le Fourn V., Girod P.A., Buceta M., Regamey A., Mermod N.
ISSN
1096-7184 (Electronic)
ISSN-L
1096-7176
Publication state
Published
Issued date
2014
Volume
21
Pages
91-102
Language
english
Abstract
The ability to efficiently produce recombinant proteins in a secreted form is highly desirable and cultured mammalian cells such as CHO cells have become the preferred host as they secrete proteins with human-like post-translational modifications. However, attempts to express high levels of particular proteins in CHO cells may consistently result in low yields, even for non-engineered proteins such as immunoglobulins. In this study, we identified the responsible faulty step at the stage of translational arrest, translocation and early processing for such a "difficult-to-express" immunoglobulin, resulting in improper cleavage of the light chain and its precipitation in an insoluble cellular fraction unable to contribute to immunoglobulin assembly. We further show that proper processing and secretion were restored by over-expressing human signal receptor protein SRP14 and other components of the secretion pathway. This allowed the expression of the difficult-to-express protein to high yields, and it also increased the production of an easy-to-express protein. Our results demonstrate that components of the secretory and processing pathways can be limiting, and that engineering of the secretory pathway may be used to improve the secretion efficiency of therapeutic proteins from CHO cells.
Keywords
CHO cells, Metabolic engineering, Protein secretion, Therapeutic proteins, Immunoglobulins
Pubmed
Web of science
Open Access
Yes
Create date
28/01/2014 13:40
Last modification date
20/08/2019 16:38
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