Multiplex secretome engineering enhances recombinant protein production and purity.

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Abstrakt

Tytuł:: Multiplex secretome engineering enhances recombinant protein production and purity.
Autorzy:: Kol S; The Novo Nordisk Foundation Center for Biosustainability, Technical University of Denmark, Building 220, Kemitorvet, 2800, Kgs. Lyngby, Denmark. .
Ley D; The Novo Nordisk Foundation Center for Biosustainability, Technical University of Denmark, Building 220, Kemitorvet, 2800, Kgs. Lyngby, Denmark.
Wulff T; The Novo Nordisk Foundation Center for Biosustainability, Technical University of Denmark, Building 220, Kemitorvet, 2800, Kgs. Lyngby, Denmark.
Decker M; The Novo Nordisk Foundation Center for Biosustainability, Technical University of Denmark, Building 220, Kemitorvet, 2800, Kgs. Lyngby, Denmark.
Arnsdorf J; The Novo Nordisk Foundation Center for Biosustainability, Technical University of Denmark, Building 220, Kemitorvet, 2800, Kgs. Lyngby, Denmark.
Schoffelen S; The Novo Nordisk Foundation Center for Biosustainability, Technical University of Denmark, Building 220, Kemitorvet, 2800, Kgs. Lyngby, Denmark.
Hansen AH; The Novo Nordisk Foundation Center for Biosustainability, Technical University of Denmark, Building 220, Kemitorvet, 2800, Kgs. Lyngby, Denmark.
Jensen TL; The Novo Nordisk Foundation Center for Biosustainability, Technical University of Denmark, Building 220, Kemitorvet, 2800, Kgs. Lyngby, Denmark.
Gutierrez JM; The Novo Nordisk Foundation Center for Biosustainability at the University of California, San Diego, School of Medicine, La Jolla, CA, 92093, USA.; Department of Bioengineering, University of California, San Diego, School of Medicine, La Jolla, CA, 92093, USA.
Chiang AWT; The Novo Nordisk Foundation Center for Biosustainability at the University of California, San Diego, School of Medicine, La Jolla, CA, 92093, USA.; Department of Pediatrics, University of California, San Diego, School of Medicine, La Jolla, CA, 92093, USA.
Masson HO; The Novo Nordisk Foundation Center for Biosustainability at the University of California, San Diego, School of Medicine, La Jolla, CA, 92093, USA.; Department of Bioengineering, University of California, San Diego, School of Medicine, La Jolla, CA, 92093, USA.
Palsson BO; The Novo Nordisk Foundation Center for Biosustainability, Technical University of Denmark, Building 220, Kemitorvet, 2800, Kgs. Lyngby, Denmark.; The Novo Nordisk Foundation Center for Biosustainability at the University of California, San Diego, School of Medicine, La Jolla, CA, 92093, USA.; Department of Bioengineering, University of California, San Diego, School of Medicine, La Jolla, CA, 92093, USA.; Department of Pediatrics, University of California, San Diego, School of Medicine, La Jolla, CA, 92093, USA.
Voldborg BG; The Novo Nordisk Foundation Center for Biosustainability, Technical University of Denmark, Building 220, Kemitorvet, 2800, Kgs. Lyngby, Denmark.
Pedersen LE; The Novo Nordisk Foundation Center for Biosustainability, Technical University of Denmark, Building 220, Kemitorvet, 2800, Kgs. Lyngby, Denmark.
Kildegaard HF; The Novo Nordisk Foundation Center for Biosustainability, Technical University of Denmark, Building 220, Kemitorvet, 2800, Kgs. Lyngby, Denmark.
Lee GM; The Novo Nordisk Foundation Center for Biosustainability, Technical University of Denmark, Building 220, Kemitorvet, 2800, Kgs. Lyngby, Denmark.; Department of Biological Sciences, KAIST, 291 Daehak-ro, Yuseong-gu, Daejeon, 305-701, Republic of Korea.
Lewis NE; The Novo Nordisk Foundation Center for Biosustainability at the University of California, San Diego, School of Medicine, La Jolla, CA, 92093, USA. .; Department of Bioengineering, University of California, San Diego, School of Medicine, La Jolla, CA, 92093, USA. .; Department of Pediatrics, University of California, San Diego, School of Medicine, La Jolla, CA, 92093, USA. .
Źródło:: Nature communications [Nat Commun] 2020 Apr 20; Vol. 11 (1), pp. 1908. Date of Electronic Publication: 2020 Apr 20.
Typ publikacji:: Journal Article; Research Support, N.I.H., Extramural; Research Support, Non-U.S. Gov't
Język:: English
Imprint Name(s):: Original Publication: [London] : Nature Pub. Group
MeSH Terms:: Metabolic Engineering/*methods
Recombinant Proteins/*biosynthesis
Animals ; Antibodies, Monoclonal/biosynthesis ; Antibodies, Monoclonal/isolation & purification ; Biological Products ; CHO Cells ; Chromatography ; Cricetulus ; Gene Knockout Techniques ; High-Throughput Nucleotide Sequencing ; Recombinant Proteins/genetics ; Recombinant Proteins/isolation & purification ; Rituximab ; Synthetic Biology
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Grant Information:: R35 GM119850 United States GM NIGMS NIH HHS
Substance Nomenclature:: 0 (Antibodies, Monoclonal)
0 (Biological Products)
0 (Recombinant Proteins)
4F4X42SYQ6 (Rituximab)
Entry Date(s):: Date Created: 20200422 Date Completed: 20200804 Latest Revision: 20220422
Update Code:: 20240105
PubMed Central ID:: PMC7170862
DOI:: 10.1038/s41467-020-15866-w
PMID:: 32313013
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Host cell proteins (HCPs) are process-related impurities generated during biotherapeutic protein production. HCPs can be problematic if they pose a significant metabolic demand, degrade product quality, or contaminate the final product. Here, we present an effort to create a "clean" Chinese hamster ovary (CHO) cell by disrupting multiple genes to eliminate HCPs. Using a model of CHO cell protein secretion, we predict that the elimination of unnecessary HCPs could have a non-negligible impact on protein production. We analyze the HCP content of 6-protein, 11-protein, and 14-protein knockout clones. These cell lines exhibit a substantial reduction in total HCP content (40%-70%). We also observe higher productivity and improved growth characteristics in specific clones. The reduced HCP content facilitates purification of a monoclonal antibody. Thus, substantial improvements can be made in protein titer and purity through large-scale HCP deletion, providing an avenue to increased quality and affordability of high-value biopharmaceuticals.

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