Proteomic profiling of CHO cells with enhanced rhBMP-2 productivity following co-expression of PACEsol.
Chinese hamster ovary (CHO) cells are widely used for the production of recombinant protein biopharmaceuticals. The purpose of this study was to investigate differences in the proteome of CHO DUKX cells expressing recombinant human bone morphogenetic protein-2 (rhBMP-2) (G5 cells) compared to cells also expressing soluble exogenous paired basic amino acid cleaving enzyme soluble paired basic amino acid cleaving enzyme (PACEsol) (3C9 cells), which has been previously found to improve the post-translational processing of the mature rhBMP-2 dimer. PACEsol co-expression was also associated with a significant increase (almost four-fold) in cellular productivity of rhBMP-2 protein. Differential proteomic expression profiling using 2-D DIGE and MALDI-TOF MS was performed to compare 3C9 and G5 cells, and revealed a list of 60 proteins that showed differential expression (up/downregulated), with a variety of different cellular functions. A substantial number of these altered proteins were found to have chaperone activity, involved with protein folding, assembly and secretion, as well as a number of proteins involved in protein translation. These results support the use of proteomic profiling as a valuable tool towards understanding the biology of bioprocess cultures.
National Institute for Cellular Biotechnology, Dublin City University, Dublin, Ireland. paula.meleady@dcu.ie
Paula
P
Meleady
Michael
M
Henry
Patrick
P
Gammell
Padraig
P
Doolan
Martin
M
Sinacore
Mark
M
Melville
Linda
L
Francullo
Mark
M
Leonard
Timothy
T
Charlebois
Martin
M
Clynes
D000818:Animals; D055396:Bone Morphogenetic Protein 2; D019485:Bone Morphogenetic Proteins / Q000737:chemistry / Q000302:isolation & purification / Q000378:metabolism*; D016466:CHO Cells / Q000166:cytology / Q000502:physiology*; D018929:Cell Culture Techniques; D002460:Cell Line; D002999:Clone Cells; D006224:Cricetinae; D003412:Cricetulus; D019281:Dimerization; D015180:Electrophoresis, Gel, Two-Dimensional; D045683:Furin / Q000235:genetics / Q000378:metabolism*; D015870:Gene Expression / Q000502:physiology*; D020869:Gene Expression Profiling; D006801:Humans; D010449:Peptide Mapping; D020543:Proteome / Q000378:metabolism; D040901:Proteomics / Q000379:methods*; D011994:Recombinant Proteins / Q000737:chemistry / Q000302:isolation & purification / Q000378:metabolism*; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization; D014162:Transfection; D016212:Transforming Growth Factor beta / Q000737:chemistry / Q000302:isolation & purification / Q000378:metabolism*
D055396:Bone Morphogenetic Protein 2; D019485:Bone Morphogenetic Proteins; D020543:Proteome; D011994:Recombinant Proteins; D016212:Transforming Growth Factor beta; C499469:recombinant human bone morphogenetic protein-2; C468731:FURIN protein, human; D045683:Furin