title: Sustained productivity in recombinant Chinese hamster ovary (CHO) cell lines: proteome analysis of the molecular basis for a process-related phenotype.
abstract: BACKGROUND
  The ability of mammalian cell lines to sustain cell specific productivity (Qp) over the full duration of bioprocess culture is a highly desirable phenotype, but the molecular basis for sustainable productivity has not been previously investigated in detail. In order to identify proteins that may be associated with a sustained productivity phenotype, we have conducted a proteomic profiling analysis of two matched pairs of monoclonal antibody-producing Chinese hamster ovary (CHO) cell lines that differ in their ability to sustain productivity over a 10 day fed-batch culture.
  
  
  RESULTS
  Proteomic profiling of inherent differences between the two sets of comparators using 2D-DIGE (Difference Gel Electrophoresis) and LC-MS/MS resulted in the identification of 89 distinct differentially expressed proteins. Overlap comparisons between the two sets of cell line pairs identified 12 proteins (AKRIB8, ANXA1, ANXA4, EIF3I, G6PD, HSPA8, HSP90B1, HSPD1, NUDC, PGAM1, RUVBL1 and CNN3) that were differentially expressed in the same direction.
  
  
  CONCLUSIONS
  These proteins may have an important role in sustaining high productivity of recombinant protein over the duration of a fed-batch bioprocess culture. It is possible that many of these proteins could be useful for future approaches to successfully manipulate or engineer CHO cells in order to sustain productivity of recombinant protein.
authors.affiliation: National Institute for Cellular Biotechnology, Dublin City University, Dublin 9, Ireland. paula.meleady@dcu.ie
authors.forename: Paula
authors.initials: P
authors.lastname: Meleady
authors.forename: Padraig
authors.initials: P
authors.lastname: Doolan
authors.forename: Michael
authors.initials: M
authors.lastname: Henry
authors.forename: Niall
authors.initials: N
authors.lastname: Barron
authors.forename: Joanne
authors.initials: J
authors.lastname: Keenan
authors.forename: Finbar
authors.initials: F
authors.lastname: O'Sullivan
authors.forename: Colin
authors.initials: C
authors.lastname: Clarke
authors.forename: Patrick
authors.initials: P
authors.lastname: Gammell
authors.forename: Mark W
authors.initials: MW
authors.lastname: Melville
authors.forename: Mark
authors.initials: M
authors.lastname: Leonard
authors.forename: Martin
authors.initials: M
authors.lastname: Clynes
mesh_terms: D000818:Animals; D000911:Antibodies, Monoclonal / Q000032:analysis / Q000378:metabolism; D001709:Biotechnology; D015153:Blotting, Western; D016466:CHO Cells; D002470:Cell Survival; D002853:Chromatography, Liquid; D006224:Cricetinae; D003412:Cricetulus; D005954:Glucosephosphate Dehydrogenase; D010641:Phenotype; D020543:Proteome / Q000032:analysis* / Q000378:metabolism; D011994:Recombinant Proteins / Q000032:analysis / Q000378:metabolism* / Q000592:standards; D053719:Tandem Mass Spectrometry; D057825:Two-Dimensional Difference Gel Electrophoresis
chemical_list: D000911:Antibodies, Monoclonal; D020543:Proteome; D011994:Recombinant Proteins; D005954:Glucosephosphate Dehydrogenase