The Expression Pattern of the Phosphoproteome Is Significantly Changed During the Growth Phases of Recombinant CHO Cell Culture.
Biotechnol J
; 13(10): e1700221, 2018 Oct.
Article
en En
| MEDLINE
| ID: mdl-30076757
Post-translational modification of proteins by reversible phosphorylation plays a pivotal role in regulating key cellular processes including transcription, translation, cell proliferation, differentiation, apoptosis, and signal transduction. Despite the importance of the phosphorylation level of regulation, little work has been carried out on the phosphoproteomic characterization of Chinese hamster ovary (CHO) cells in bioprocess-relevant conditions. Growth control strategies are often used to prolong culture duration and increase specific productivity; however, the cellular mechanisms and regulatory pathways underlying growth strategies are poorly understood in CHO cells. Phosphorylation changes are dynamic and will respond to changes in culture conditions; this may reflect the status of the cells with respect to growth and viability of the culture. Herein, this study uses a phosphopeptide enrichment strategy in conjunction with LC-MS/MS to carry out a large-scale differential phosphoproteomic analysis of IgG producing CHO DP12 cells at various phases of growth in serum-free suspension batch culture to characterize dynamic changes to the phosphoproteome with changing culture conditions. In total over the various growth phases, 3777 differentially expressed unique phosphopeptides are identified from 1415 differentially expressed unique phosphoproteins. Analysis of the whole cell lysate without phosphopeptide enrichment over the various growth phases revealed the differential expression of 834 unique proteins, with an overlap of 188 proteins between the proteomic and phosphoproteomic analyses. The inclusion of phosphoproteomic data significantly improves proteome coverage but also gives insights into the post-translational level of regulation during cellular growth of recombinant CHO cells.
Palabras clave
Texto completo:
1
Colección:
01-internacional
Base de datos:
MEDLINE
Asunto principal:
Fosfoproteínas
/
Proteoma
/
Proliferación Celular
Límite:
Animals
Idioma:
En
Revista:
Biotechnol J
Asunto de la revista:
BIOTECNOLOGIA
Año:
2018
Tipo del documento:
Article
País de afiliación:
Irlanda
Pais de publicación:
Alemania