title: High level expression and purification of recombinant PEX protein in cultured skeletal muscle cell expression system.
abstract: Large quantities of recombinant proteins are needed for specific therapeutic and diagnostic applications. To achieve high-level expression in eukaryotic cells, the choice of cell line as well as the expression vector is critical. In this report, we demonstrate that a combination of the skeletal muscle cell line, QM7 and a cytomegalovirus promoter-based expression vector can achieve high-level expression of secretory recombinant proteins in eukaryotic cells. We also screened a serum-free medium containing 3 microg/ml insulin suitable for QM7 differentiation and identified a very potent signal peptide from MMP9, which effectively directs secretion of heterologous proteins. The C-terminal hemopexin-like domain of MMP-2, PEX, a powerful candidate for the treatment of diseases associated with neovascularization was expressed in QM7 cells with bioactivity. This skeletal muscle cell-based system may be employed for the production of human proteins of special interests, such as those for structural determination or therapeutical development.
authors.affiliation: Department of Cell Biology, Peking University School of Oncology, Beijing Cancer Hospital & Institute, Beijing 100036, China.
authors.forename: Lin
authors.initials: L
authors.lastname: Song
authors.forename: Yang
authors.initials: Y
authors.lastname: Ke
authors.forename: Zhi-Qian
authors.initials: ZQ
authors.lastname: Zhang
mesh_terms: D000818:Animals; D015870:Gene Expression / Q000502:physiology; D018485:Muscle Fibers, Skeletal / Q000378:metabolism*; D018482:Muscle, Skeletal / Q000378:metabolism*; D053402:PHEX Phosphate Regulating Neutral Endopeptidase / Q000096:biosynthesis* / Q000737:chemistry / Q000235:genetics / Q000302:isolation & purification*; D015202:Protein Engineering / Q000379:methods*; D011784:Quail; D011994:Recombinant Proteins / Q000096:biosynthesis* / Q000737:chemistry / Q000302:isolation & purification*
chemical_list: D011994:Recombinant Proteins; D053402:PHEX Phosphate Regulating Neutral Endopeptidase; C506231:PHEX protein, human