title: Recombinant murine growth hormone from E. coli inclusion bodies: expression, high-pressure solubilization and refolding, and characterization of activity and structure.
abstract: We expressed recombinant murine growth hormone (rmGH) in E. coli as a cost-effective way to produce large quantities (gram scale) of the protein for use in murine studies of immunogenicity to therapeutic proteins. High hydrostatic pressure was used to achieve high solubility and high refolding yields of rmGH protein produced in E. coli inclusion bodies. A two-step column purification protocol was used to produce 99% pure monomeric rmGH. Secondary and tertiary structures of purified rmGH were investigated using circular dichroism and 2D-UV spectroscopy. The purified rmGH produced was found to be biologically active in hypophysectomized rats.
authors.affiliation: Dept. of Chemical Engineering, University of Colorado at Boulder, Boulder, CO 80309, USA.
authors.forename: Amber Haynes
authors.initials: AH
authors.lastname: Fradkin
authors.forename: Carl S
authors.initials: CS
authors.lastname: Boand
authors.forename: Stephen P
authors.initials: SP
authors.lastname: Eisenberg
authors.forename: Mary S
authors.initials: MS
authors.lastname: Rosendahl
authors.forename: Theodore W
authors.initials: TW
authors.lastname: Randolph
mesh_terms: D000818:Animals; D002850:Chromatography, Gel; D002852:Chromatography, Ion Exchange; D002942:Circular Dichroism; D004591:Electrophoresis, Polyacrylamide Gel; D004926:Escherichia coli / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism; D005285:Fermentation; D013006:Growth Hormone / Q000096:biosynthesis* / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism; D007016:Hypophysectomy; D002479:Inclusion Bodies / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism; D008297:Male; D051379:Mice; D017510:Protein Folding; D051381:Rats; D017207:Rats, Sprague-Dawley; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D012995:Solubility
chemical_list: D011994:Recombinant Proteins; D013006:Growth Hormone