Dataset Information

Time-resolved proteomic profiling for CHO-K1 fed-batch cultivations

ABSTRACT: The majority of recombinant protein therapeutics are produced with Chinese Hamster Ovary (CHO) cells. Productivity depends on the initial cell line engineering in terms of integration site or choice of an appropriate promotor for the recombinant gene expression, as well as media and process parameter optimization. Here, proteomic profiling is used to identify optimization targets for a pharmaceutical relevant cell line system. Triplicates of CHO-K1 cell 2 L bioreactor fedbatch cultivations were performed and daily sampled for nLC-MS/MS proteomic analysis. Collected data from day 3 up to day 11 showed high Pearson correlation of 93.7 ± 4 % with ca. 2500 proteins quantified. The different growth phases were separated by principal component analysis and hierarchical clustering approaches. Subsequent statistical analysis revealed distinct protein profiles, where steady increase or decrease over time were the most prominent clusters. Fisher exact enrichment tests yielded in significantly enriched protein annotations which were successfully mapped to growth and metabolic changes during fedbatch cell cultivation. Major improvements in cellular and process understanding were achieved and yielded in the identification of promising new targets, like strong endogenous promotors, for cellular engineering and process optimization of this biopharmaceutical relevant cell line.

INSTRUMENT(S):

ORGANISM(S): Cricetulus Griseus (chinese Hamster) (cricetulus Barabensis Griseus)

TISSUE(S): Permanent Cell Line Cell, Cell Culture

SUBMITTER: Louise Schelletter