Multiplex secretome engineering enhances recombinant protein production and purity.

Animals Antibodies, Monoclonal / biosynthesis Biological Products CHO Cells Chromatography Cricetulus Gene Knockout Techniques High-Throughput Nucleotide Sequencing Metabolic Engineering / methods Recombinant Proteins / biosynthesis Rituximab Synthetic Biology

Journal

Nature communications
ISSN: 2041-1723
Titre abrégé: Nat Commun
Pays: England
ID NLM: 101528555

Informations de publication

Date de publication:
20 04 2020
Historique:
received: 22 05 2019
accepted: 31 03 2020
entrez: 22 4 2020
pubmed: 22 4 2020
medline: 5 8 2020
Statut: epublish

Résumé

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.

Identifiants

DOI: 10.1038/s41467-020-15866-w PMID: 32313013 PMC: PMC7170862
pubmed: 32313013
doi: 10.1038/s41467-020-15866-w
pii: 10.1038/s41467-020-15866-w
pmc: PMC7170862
doi:

Substances chimiques

Antibodies, Monoclonal 0
Biological Products 0
Recombinant Proteins 0
Rituximab 4F4X42SYQ6

Types de publication

Journal Article Research Support, N.I.H., Extramural Research Support, Non-U.S. Gov't

Langues

eng

Sous-ensembles de citation

IM

Pagination

1908

Subventions

Organisme : NIGMS NIH HHS
ID : R35 GM119850
Pays : United States

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Auteurs

Stefan Kol (S)

The Novo Nordisk Foundation Center for Biosustainability, Technical University of Denmark, Building 220, Kemitorvet, 2800, Kgs. Lyngby, Denmark. stef.kol@gmail.com.
ORCID: 0000-0002-1992-8242

Daniel Ley (D)

The Novo Nordisk Foundation Center for Biosustainability, Technical University of Denmark, Building 220, Kemitorvet, 2800, Kgs. Lyngby, Denmark.

Tune Wulff (T)

The Novo Nordisk Foundation Center for Biosustainability, Technical University of Denmark, Building 220, Kemitorvet, 2800, Kgs. Lyngby, Denmark.

Marianne Decker (M)

The Novo Nordisk Foundation Center for Biosustainability, Technical University of Denmark, Building 220, Kemitorvet, 2800, Kgs. Lyngby, Denmark.

Johnny Arnsdorf (J)

The Novo Nordisk Foundation Center for Biosustainability, Technical University of Denmark, Building 220, Kemitorvet, 2800, Kgs. Lyngby, Denmark.

Sanne Schoffelen (S)

The Novo Nordisk Foundation Center for Biosustainability, Technical University of Denmark, Building 220, Kemitorvet, 2800, Kgs. Lyngby, Denmark.
ORCID: 0000-0003-2664-8561

Anders Holmgaard Hansen (AH)

The Novo Nordisk Foundation Center for Biosustainability, Technical University of Denmark, Building 220, Kemitorvet, 2800, Kgs. Lyngby, Denmark.
ORCID: 0000-0002-6007-4787

Tanja Lyholm Jensen (TL)

The Novo Nordisk Foundation Center for Biosustainability, Technical University of Denmark, Building 220, Kemitorvet, 2800, Kgs. Lyngby, Denmark.

Jahir M 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.

Austin W T 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.
ORCID: 0000-0001-6514-5612

Helen O 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.

Bernhard O 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.

Bjørn G Voldborg (BG)

The Novo Nordisk Foundation Center for Biosustainability, Technical University of Denmark, Building 220, Kemitorvet, 2800, Kgs. Lyngby, Denmark.

Lasse Ebdrup Pedersen (LE)

The Novo Nordisk Foundation Center for Biosustainability, Technical University of Denmark, Building 220, Kemitorvet, 2800, Kgs. Lyngby, Denmark.
ORCID: 0000-0002-6064-919X

Helene Faustrup Kildegaard (HF)

The Novo Nordisk Foundation Center for Biosustainability, Technical University of Denmark, Building 220, Kemitorvet, 2800, Kgs. Lyngby, Denmark.

Gyun Min 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.
ORCID: 0000-0002-9876-7378

Nathan E Lewis (NE)

The Novo Nordisk Foundation Center for Biosustainability at the University of California, San Diego, School of Medicine, La Jolla, CA, 92093, USA. nlewisres@ucsd.edu.
Department of Bioengineering, University of California, San Diego, School of Medicine, La Jolla, CA, 92093, USA. nlewisres@ucsd.edu.
Department of Pediatrics, University of California, San Diego, School of Medicine, La Jolla, CA, 92093, USA. nlewisres@ucsd.edu.
ORCID: 0000-0001-7700-3654

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Classifications MeSH

questionsmedicales.fr › Eucaryotes › Animaux › Multiplex secretome engineering enhances...
questionsmedicales.fr › Acides aminés, peptides et protéines › Protéines › Globulines › Sérum-globulines › Immunoglobulines › Anticorps › Anticorps monoclonaux › Multiplex secretome engineering enhances...
questionsmedicales.fr › Mélanges complexes › Produits biologiques › Multiplex secretome engineering enhances...
questionsmedicales.fr › Cellules › Cellules épithéliales › Cellules CHO › Multiplex secretome engineering enhances...
questionsmedicales.fr › Techniques d'investigation › Techniques de chimie analytique › Chromatographie › Multiplex secretome engineering enhances...
questionsmedicales.fr › Eucaryotes › Animaux › Chordés › Vertébrés › Mammifères › Eutheria › Rodentia › Muridae › Arvicolinae › Cricetinae › Cricetulus › Multiplex secretome engineering enhances...
questionsmedicales.fr › Techniques d'investigation › Techniques génétiques › Ciblage de gène › Techniques de knock-out de gènes › Multiplex secretome engineering enhances...
questionsmedicales.fr › Techniques d'investigation › Techniques génétiques › Analyse de séquence › Séquençage nucléotidique à haut débit › Multiplex secretome engineering enhances...
questionsmedicales.fr › Technologie, industrie et agriculture › Ingénierie › Bioingénierie › Ingénierie cellulaire › Génie métabolique › Multiplex secretome engineering enhances...
questionsmedicales.fr › Acides aminés, peptides et protéines › Protéines › Protéines recombinantes › Multiplex secretome engineering enhances...
questionsmedicales.fr › Acides aminés, peptides et protéines › Protéines › Globulines › Sérum-globulines › Immunoglobulines › Anticorps › Anticorps monoclonaux › Anticorps monoclonaux d'origine murine › Rituximab › Multiplex secretome engineering enhances...
questionsmedicales.fr › Technologie, industrie et agriculture › Ingénierie › Bioingénierie › Biologie synthétique › Multiplex secretome engineering enhances...