LC-MS/MS-based quantitative proteomic and phosphoproteomic analysis of CHO-K1 cells adapted to growth in glutamine-free
- PDF / 1,011,152 Bytes
- 14 Pages / 547.087 x 737.008 pts Page_size
- 3 Downloads / 167 Views
(0123456789().,-volV) ( 01234567 89().,-volV)
ORIGINAL RESEARCH PAPER
LC-MS/MS-based quantitative proteomic and phosphoproteomic analysis of CHO-K1 cells adapted to growth in glutamine-free media Prashant Kaushik . Ricardo Valde´s-Bango Curell . Michael Henry . Niall Barron . Paula Meleady
Received: 26 March 2020 / Accepted: 28 June 2020 Ó Springer Nature B.V. 2020
Abstract Objectives This study aims to provide insights into the molecular mechanisms underlying adaptation of CHO-K1 cells to growth in glutamine-free media and potentially identifying critical signalling proteins and pathways involved in this phenotype. Results A CHO-K1 cell line adapted to growth in glutamine-free media was established using a straightforward one-step glutamine reduction strategy. The adapted cell line had a comparable phenotype to the parental cells in terms of cell growth and viability. Global quantitative proteomic and phosphoproteomic
Prashant Kaushik and Ricardo Valde´s-Bango Curell contributed equally to the work.
Electronic supplementary material The online version of this article (https://doi.org/10.1007/s10529-020-02953-7) contains supplementary material, which is available to authorized users. P. Kaushik R. V.-B. Curell M. Henry P. Meleady (&) National Institute for Cellular Biotechnology, Dublin City University, Dublin 9, Ireland e-mail: [email protected]
analysis was carried out to compare the cells adapted to growth in glutamine-free media to parental cells grown in media containing 8 mM L-glutamine. The adaptation process was accompanied by changes in proteins associated with cytoskeleton rearrangement and mRNA splicing as evidenced via functional analysis of 194 differentially expressed proteins between the two cell lines. 434 phosphoproteins with altered abundance were also identified as a result of adaptation to L-glutamine-free conditions with an associated enrichment of pathways associated with MAPK and calcium signalling. Conclusions This work provides a comprehensive proteomic and phosphoproteomic analysis of protein expression changes after adaptation to glutamine-free growth conditions highlighting critical pathways to consider in the rational design of improved feeding strategies or in cell line engineering to improve bioprocess phenotypes. Keywords Chinese hamster ovary cells Glutamine metabolism Phosphoproteomics Site-specific phosphorylation Phosphopeptide enrichment Biopharmaceuticals
N. Barron National Institute for Bioprocessing Research and Training, Dublin 4, Ireland N. Barron School of Chemical and Bioprocess Engineering, University College Dublin, Belfield, Dublin 4, Ireland
123
Biotechnol Lett
Introduction Chinese hamster ovary (CHO) cells are currently the primary host cell line used to produce biopharmaceuticals, and much work is being carried out to improve efficiency of production to increase product titre and improve product quality. Up to 85% of new monoclonal antibody (Mab) products approved between 2014 and 2018 were produced in Chinese hamster ovary (CHO) cells
Data Loading...
