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Mapping the molecular basis for growth related phenotypes in industrial producer CHO cell lines using differential proteomic analysis
BACKGROUND: The ability to achieve high peak viable cell density earlier in CHO cell culture and maintain an extended cell viability throughout the production process is highly desirable to increase recombinant protein yields, reduce host cell impurities for downstream processing and reduce the cost...
Autores principales: | , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
BioMed Central
2021
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8305936/ https://www.ncbi.nlm.nih.gov/pubmed/34301236 http://dx.doi.org/10.1186/s12896-021-00704-8 |
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author | Bryan, Laura Henry, Michael Kelly, Ronan M. Frye, Christopher C. Osborne, Matthew D. Clynes, Martin Meleady, Paula |
author_facet | Bryan, Laura Henry, Michael Kelly, Ronan M. Frye, Christopher C. Osborne, Matthew D. Clynes, Martin Meleady, Paula |
author_sort | Bryan, Laura |
collection | PubMed |
description | BACKGROUND: The ability to achieve high peak viable cell density earlier in CHO cell culture and maintain an extended cell viability throughout the production process is highly desirable to increase recombinant protein yields, reduce host cell impurities for downstream processing and reduce the cost of goods. In this study we implemented label-free LC-MS/MS proteomic profiling of IgG4 producing CHO cell lines throughout the duration of the cell culture to identify differentially expressed (DE) proteins and intracellular pathways associated with the high peak viable cell density (VCD) and extended culture VCD phenotypes. RESULTS: We identified key pathways in DNA replication, mitotic cell cycle and evasion of p53 mediated apoptosis in high peak VCD clonally derived cell lines (CDCLs). ER to Golgi vesicle mediated transport was found to be highly expressed in extended culture VCD CDCLs while networks involving endocytosis and oxidative stress response were significantly downregulated. CONCLUSION: This investigation highlights key pathways for targeted engineering to generate desirable CHO cell phenotypes for biotherapeutic production. SUPPLEMENTARY INFORMATION: The online version contains supplementary material available at 10.1186/s12896-021-00704-8. |
format | Online Article Text |
id | pubmed-8305936 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2021 |
publisher | BioMed Central |
record_format | MEDLINE/PubMed |
spelling | pubmed-83059362021-07-28 Mapping the molecular basis for growth related phenotypes in industrial producer CHO cell lines using differential proteomic analysis Bryan, Laura Henry, Michael Kelly, Ronan M. Frye, Christopher C. Osborne, Matthew D. Clynes, Martin Meleady, Paula BMC Biotechnol Research BACKGROUND: The ability to achieve high peak viable cell density earlier in CHO cell culture and maintain an extended cell viability throughout the production process is highly desirable to increase recombinant protein yields, reduce host cell impurities for downstream processing and reduce the cost of goods. In this study we implemented label-free LC-MS/MS proteomic profiling of IgG4 producing CHO cell lines throughout the duration of the cell culture to identify differentially expressed (DE) proteins and intracellular pathways associated with the high peak viable cell density (VCD) and extended culture VCD phenotypes. RESULTS: We identified key pathways in DNA replication, mitotic cell cycle and evasion of p53 mediated apoptosis in high peak VCD clonally derived cell lines (CDCLs). ER to Golgi vesicle mediated transport was found to be highly expressed in extended culture VCD CDCLs while networks involving endocytosis and oxidative stress response were significantly downregulated. CONCLUSION: This investigation highlights key pathways for targeted engineering to generate desirable CHO cell phenotypes for biotherapeutic production. SUPPLEMENTARY INFORMATION: The online version contains supplementary material available at 10.1186/s12896-021-00704-8. BioMed Central 2021-07-23 /pmc/articles/PMC8305936/ /pubmed/34301236 http://dx.doi.org/10.1186/s12896-021-00704-8 Text en © The Author(s) 2021 https://creativecommons.org/licenses/by/4.0/Open AccessThis article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons licence, and indicate if changes were made. The images or other third party material in this article are included in the article's Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article's Creative Commons licence and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this licence, visit http://creativecommons.org/licenses/by/4.0/ (https://creativecommons.org/licenses/by/4.0/) . The Creative Commons Public Domain Dedication waiver (http://creativecommons.org/publicdomain/zero/1.0/ (https://creativecommons.org/publicdomain/zero/1.0/) ) applies to the data made available in this article, unless otherwise stated in a credit line to the data. |
spellingShingle | Research Bryan, Laura Henry, Michael Kelly, Ronan M. Frye, Christopher C. Osborne, Matthew D. Clynes, Martin Meleady, Paula Mapping the molecular basis for growth related phenotypes in industrial producer CHO cell lines using differential proteomic analysis |
title | Mapping the molecular basis for growth related phenotypes in industrial producer CHO cell lines using differential proteomic analysis |
title_full | Mapping the molecular basis for growth related phenotypes in industrial producer CHO cell lines using differential proteomic analysis |
title_fullStr | Mapping the molecular basis for growth related phenotypes in industrial producer CHO cell lines using differential proteomic analysis |
title_full_unstemmed | Mapping the molecular basis for growth related phenotypes in industrial producer CHO cell lines using differential proteomic analysis |
title_short | Mapping the molecular basis for growth related phenotypes in industrial producer CHO cell lines using differential proteomic analysis |
title_sort | mapping the molecular basis for growth related phenotypes in industrial producer cho cell lines using differential proteomic analysis |
topic | Research |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8305936/ https://www.ncbi.nlm.nih.gov/pubmed/34301236 http://dx.doi.org/10.1186/s12896-021-00704-8 |
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