Mapping the molecular basis for growth related phenotypes in industrial producer CHO cell lines using differential proteomic analysis

Laura Bryan,Christopher C Frye,Ronan M Kelly,Martin Clynes,Matthew D Osborne,Michael Henry,Paula Meleady

doi:10.1186/s12896-021-00704-8

Laura Bryan, Christopher C Frye + Show 5 more

Open Access

https://doi.org/10.1186/s12896-021-00704-8

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Abstract

BackgroundThe 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.ResultsWe 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.ConclusionThis investigation highlights key pathways for targeted engineering to generate desirable CHO cell phenotypes for biotherapeutic production.

Highlights

The ability to achieve high peak viable cell density earlier in Chinese hamster ovary (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
Waste products and metabolites of the cells were measured throughout culture with no significant difference in lactate or ammonia being detected between high and low peak viable cell density (VCD) clonally derived cell lines (CDCLs) (Additional file 3)
Both phenotypes investigated are inherently related to the growth of the cell, differential LC-MS/MS proteomic analysis revealed different pathways and biological processes that are significantly enriched in each experiment

Summary

Introduction

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. CHO cell lines that reach high peak VCDs early in culture are desirable due to their potential to reduce cell culture production duration and allow for increased seeding densities. These improvements in the efficiency of production will lead to a reduction in the costs of complex biotherapeutics making them more accessible to patients. The IVCD of the culture has been shown to be positively correlated with product titre [41]

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