Recombinant murine growth hormone from E. coli inclusion bodies: Expression, high-pressure solubilization and refolding, and characterization of activity and structure
| Autor: | Amber Haynes Fradkin, Stephen P. Eisenberg, Mary S. Rosendahl, Carl S. Boand, Theodore W. Randolph |
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| Rok vydání: | 2010 |
| Předmět: |
Male
Protein Folding Circular dichroism Hydrostatic pressure Inclusion bodies law.invention Rats Sprague-Dawley Mice chemistry.chemical_compound law Escherichia coli Animals Solubility Hypophysectomy Inclusion Bodies Chromatography Chemistry Circular Dichroism Immunogenicity Biological activity Chromatography Ion Exchange Recombinant Proteins Rats Monomer Biochemistry Growth Hormone Fermentation Chromatography Gel Recombinant DNA Electrophoresis Polyacrylamide Gel Biotechnology |
| Zdroj: | Biotechnology Progress. 26:743-749 |
| ISSN: | 1520-6033 8756-7938 |
| DOI: | 10.1002/btpr.393 |
| Popis: | 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. |
| Databáze: | OpenAIRE |
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