title: Engineering the chaperone network of CHO cells for optimal recombinant protein production and authenticity.
abstract: All proteins fold into a defined three-dimensional shape that is compatible with the cellular role and/or biological activity of those proteins. Molecular chaperones are a family of proteins whose role is to assist the folding and targeting of proteins in both normal and stressed cells. The rational manipulation of chaperone levels in a cell line engineered to produce a defined recombinant protein (rP) can significantly improve both the achievable steady-state levels and authenticity of a wide range of recombinant proteins. Here, we describe the methodology associated with expressing a variety of molecular chaperones in Chinese hamster ovary (CHO) lines in order to improve their recombinant protein production capacity. These chaperones include both those that facilitate the folding of the polypeptide chain (i.e. Hsp70, Hsp40) and those that can re-fold proteins that have misfolded in the cell (i.e. ClpB/Hsp104). This latter property is particularly important given the propensity for highly expressed recombinant proteins to misfold in the "foreign" cellular environment.
authors.affiliation: University of Kent, Kent, UK.
authors.forename: Lyne
authors.initials: L
authors.lastname: Jossé
authors.forename: C Mark
authors.initials: CM
authors.lastname: Smales
authors.forename: Mick F
authors.initials: MF
authors.lastname: Tuite
mesh_terms: D000818:Animals; D001709:Biotechnology / Q000379:methods*; D015153:Blotting, Western; D016466:CHO Cells; D006224:Cricetinae; D003412:Cricetulus; D004591:Electrophoresis, Polyacrylamide Gel; D005818:Genetic Engineering / Q000379:methods*; D006360:Heat-Shock Proteins / Q000235:genetics / Q000378:metabolism; D008156:Luciferases; D018832:Molecular Chaperones / Q000378:metabolism*; D017510:Protein Folding; D011994:Recombinant Proteins / Q000378:metabolism*; D014162:Transfection / Q000379:methods
chemical_list: D006360:Heat-Shock Proteins; D018832:Molecular Chaperones; D011994:Recombinant Proteins; D008156:Luciferases