Proteomics of Biopharmaceutical Production Cells

Very little is known about the cellular and molecular mechanisms that govern the production of complex proteins by engineered mammalian cells. The majority of the work to date has largely concentrated on optimisation of cell culture conditions (e.g. media formulation), bioreactor design, and improvements in the design of expression vectors. We are undertaking proteomic studies on CHO cells and other industrially relevant cell lines, in order to understand the molecular and cellular mechanisms involved in growth and productivity in mammalian cells used for biopharmaceutical production. Protein profiles generated from both intracellular and extracellular samples from bioreactors may give insights for genetic intervention to possibly create better host cell lines, or even to provide clues to more rational strategies for process development (e.g. media design).

We are therefore undertaking differential proteomic and phospho-proteomic expression profiling experiments using quantitative label-free LC-MS approaches to identify and characterize specific proteins to make the production process more efficient, and in particular, to improve the growth and productivity of the cells which underlie the process.

Contact

Paula Meleady (paula.meleady@dcu.ie)

Click here to meet our CHO research group

 

Publications

  • Meleady P, Gallagher M, Clarke C, Henry M, Sanchez N, Barron N, Clynes M (2012). Impact of miR-7 over-expression on the proteome of Chinese hamster ovary cells. J Biotechnol. 160; 251-262.

 

  • Meleady P, Hoffrogge R, Henry M, Rupp O, Bort JH, Clarke C, Brinkrolf K, Kelly S, Müller B, Doolan P, Hackl M, Beckmann TF, Noll T, Grillari J, Barron N, Pühler A, Clynes M, Borth N. (2012). Utilization and evaluation of CHO-specific sequence databases for mass spectrometry based proteomics. Biotechnol Bioeng. 109:1386-94.

 

  • Meleady P*, Gallagher M*, Clarke C, Henry M, Sanchez N, Barron N, Clynes M. (2012) Impact of miR-7 over-expression on the proteome of Chinese hamster ovary cells. J Biotechnol. Aug 31;160(3- 4):251-62. *joint first author

 

  • Meleady P*, Hoffrogge R*, Henry M, Rupp O, Bort JH, Clarke C, Brinkrolf K, Kelly S, Müller B, Doolan P, Hackl M, Beckmann TF, Noll T, Grillari J, Barron N, Pühler A, Clynes M, Borth N. (2012) Utilization and evaluation of CHO-specific sequence databases for mass spectrometry based proteomics. Biotechnol Bioeng. Jun;109(6):1386-94. *joint first author

 

  • Meleady P, Doolan P, Henry M, Barron N, Keenan J, O'Sullivan F, Clarke C, Gammell P, Melville M, Leonard M, Clynes M. (2011) Sustained productivity in recombinant Chinese Hamster Ovary (CHO) cell lines: proteome analysis of the molecular basis for a process-related phenotype. BMC Biotechnol. Jul 24, 11:78.

 

  • Meleady P, Doolan P, Henry M, Barron N, Keenan J, O'Sullivan F, Clarke C, Gammell P, Melville MW, Leonard M, Clynes M (2011). Sustained productivity in recombinant Chinese hamster ovary (CHO) cell lines: proteome analysis of the molecular basis for a process-related phenotype. BMC Biotechnol. 2011 Jul 24;11:78.

 

  • Meleady P, Henry M, Gammell P, Doolan P, Sinacore M, Melville M, Francullo L, Leonard M, Charlebois T, Clynes M (2008) Proteomic profiling of CHO cells with enhanced rhBMP-2 productivity following co-expression of PACEsol. Proteomics. 8(13):2611-24

 

  • Kumar N, Gammell P, Meleady P, Henry M, Clynes M (2008) Differential protein expression following low temperature culture of suspension CHO-K1 cells. BMC Biotechnol. Apr 22; 8:42.
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