Identification of cellular signatures associated with chinese hamster ovary cell adaptation for secretion of antibodies

The secretory capacity of Chinese hamster ovary (CHO) cells remains a fundamental bottleneck in the manufacturing of protein-based therapeutics. Unconventional biological drugs with complex structures and processing requirements are particularly problematic. Although engineered vector DNA elements c...

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Main Authors: Ying Bai, Ivan Domenech Mercadé, Ramy Elgendy, Giulia Lambiase, Sew Peak-Chew, Catarina Franco, Steven W. Wingett, Tim J. Stevens, Luigi Grassi, Noah Hitchcock, Cristina Sayago Ferreira, Diane Hatton, Elizabeth A. Miller, Rajesh K. Mistry
Format: Article
Language:English
Published: Elsevier 2025-01-01
Series:Computational and Structural Biotechnology Journal
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Online Access:http://www.sciencedirect.com/science/article/pii/S200103702400429X
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author Ying Bai
Ivan Domenech Mercadé
Ramy Elgendy
Giulia Lambiase
Sew Peak-Chew
Catarina Franco
Steven W. Wingett
Tim J. Stevens
Luigi Grassi
Noah Hitchcock
Cristina Sayago Ferreira
Diane Hatton
Elizabeth A. Miller
Rajesh K. Mistry
author_facet Ying Bai
Ivan Domenech Mercadé
Ramy Elgendy
Giulia Lambiase
Sew Peak-Chew
Catarina Franco
Steven W. Wingett
Tim J. Stevens
Luigi Grassi
Noah Hitchcock
Cristina Sayago Ferreira
Diane Hatton
Elizabeth A. Miller
Rajesh K. Mistry
author_sort Ying Bai
collection DOAJ
description The secretory capacity of Chinese hamster ovary (CHO) cells remains a fundamental bottleneck in the manufacturing of protein-based therapeutics. Unconventional biological drugs with complex structures and processing requirements are particularly problematic. Although engineered vector DNA elements can achieve rapid and high-level therapeutic protein production, a high metabolic and protein folding burden is imposed on the host cell. Cellular adaptations to these conditions include differential gene expression profiles that can in turn influence the productivity and quality control of recombinant proteins. In this study, we used quantitative transcriptomic and proteomic analyses to investigate how biological pathways change with antibody titre. Gene and protein expression profiles of CHO cell pools and clones producing a panel of different monoclonal and bispecific antibodies were analysed during fed-batch production. Antibody-expressing CHO cell pools were heterogeneous, resulting in few discernible genetic signatures. Clonal cell lines derived from these pools, selected for high and low production, yielded a small number of differentially expressed proteins that correlated with productivity and were shared across the biotherapeutics. However, the dominant feature associated with higher protein production was transgene copy number and the resulting mRNA expression level. Moreover, variability between clonal cell lines suggested that the process of cellular adaptation is variable with diverse cellular changes associated with individual adaptation events.
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spelling doaj-art-808e407dcb674f72af2e309420f393fc2025-08-20T01:59:30ZengElsevierComputational and Structural Biotechnology Journal2001-03702025-01-0127173110.1016/j.csbj.2024.12.006Identification of cellular signatures associated with chinese hamster ovary cell adaptation for secretion of antibodiesYing Bai0Ivan Domenech Mercadé1Ramy Elgendy2Giulia Lambiase3Sew Peak-Chew4Catarina Franco5Steven W. Wingett6Tim J. Stevens7Luigi Grassi8Noah Hitchcock9Cristina Sayago Ferreira10Diane Hatton11Elizabeth A. Miller12Rajesh K. Mistry13Cell Biology Division, Medical Research Council Laboratory of Molecular Biology, Cambridge CB2 0QH, UK; Cell Culture and Fermentation Sciences, BioPharmaceutical Development, AstraZeneca, Cambridge UKCell Biology Division, Medical Research Council Laboratory of Molecular Biology, Cambridge CB2 0QH, UK; Cell Culture and Fermentation Sciences, BioPharmaceutical Development, AstraZeneca, Cambridge UKTranslational Genomics, Discovery Sciences, BioPharmaceutical R&D, AstraZeneca, Gothenburg, SwedenAnalytical Sciences, BioPharmaceutical Development, AstraZeneca, Cambridge, UKCell Biology Division, Medical Research Council Laboratory of Molecular Biology, Cambridge CB2 0QH, UKCell Biology Division, Medical Research Council Laboratory of Molecular Biology, Cambridge CB2 0QH, UKCell Biology Division, Medical Research Council Laboratory of Molecular Biology, Cambridge CB2 0QH, UKCell Biology Division, Medical Research Council Laboratory of Molecular Biology, Cambridge CB2 0QH, UKCell Culture and Fermentation Sciences, BioPharmaceutical Development, AstraZeneca, Cambridge UKCell Culture and Fermentation Sciences, BioPharmaceutical Development, AstraZeneca, Cambridge UKCell Culture and Fermentation Sciences, BioPharmaceutical Development, AstraZeneca, Cambridge UKCell Culture and Fermentation Sciences, BioPharmaceutical Development, AstraZeneca, Cambridge UKCell Biology Division, Medical Research Council Laboratory of Molecular Biology, Cambridge CB2 0QH, UK; Corresponding authors.Cell Culture and Fermentation Sciences, BioPharmaceutical Development, AstraZeneca, Cambridge UK; Corresponding authors.The secretory capacity of Chinese hamster ovary (CHO) cells remains a fundamental bottleneck in the manufacturing of protein-based therapeutics. Unconventional biological drugs with complex structures and processing requirements are particularly problematic. Although engineered vector DNA elements can achieve rapid and high-level therapeutic protein production, a high metabolic and protein folding burden is imposed on the host cell. Cellular adaptations to these conditions include differential gene expression profiles that can in turn influence the productivity and quality control of recombinant proteins. In this study, we used quantitative transcriptomic and proteomic analyses to investigate how biological pathways change with antibody titre. Gene and protein expression profiles of CHO cell pools and clones producing a panel of different monoclonal and bispecific antibodies were analysed during fed-batch production. Antibody-expressing CHO cell pools were heterogeneous, resulting in few discernible genetic signatures. Clonal cell lines derived from these pools, selected for high and low production, yielded a small number of differentially expressed proteins that correlated with productivity and were shared across the biotherapeutics. However, the dominant feature associated with higher protein production was transgene copy number and the resulting mRNA expression level. Moreover, variability between clonal cell lines suggested that the process of cellular adaptation is variable with diverse cellular changes associated with individual adaptation events.http://www.sciencedirect.com/science/article/pii/S200103702400429XMultiomicsChinese hamster ovary cellsBiotherapeutics
spellingShingle Ying Bai
Ivan Domenech Mercadé
Ramy Elgendy
Giulia Lambiase
Sew Peak-Chew
Catarina Franco
Steven W. Wingett
Tim J. Stevens
Luigi Grassi
Noah Hitchcock
Cristina Sayago Ferreira
Diane Hatton
Elizabeth A. Miller
Rajesh K. Mistry
Identification of cellular signatures associated with chinese hamster ovary cell adaptation for secretion of antibodies
Computational and Structural Biotechnology Journal
Multiomics
Chinese hamster ovary cells
Biotherapeutics
title Identification of cellular signatures associated with chinese hamster ovary cell adaptation for secretion of antibodies
title_full Identification of cellular signatures associated with chinese hamster ovary cell adaptation for secretion of antibodies
title_fullStr Identification of cellular signatures associated with chinese hamster ovary cell adaptation for secretion of antibodies
title_full_unstemmed Identification of cellular signatures associated with chinese hamster ovary cell adaptation for secretion of antibodies
title_short Identification of cellular signatures associated with chinese hamster ovary cell adaptation for secretion of antibodies
title_sort identification of cellular signatures associated with chinese hamster ovary cell adaptation for secretion of antibodies
topic Multiomics
Chinese hamster ovary cells
Biotherapeutics
url http://www.sciencedirect.com/science/article/pii/S200103702400429X
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