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Recombinant murine growth hormone from E. coli inclusion bodies: Expression, high‐pressure solubilization and refolding, and characterization of activity and structure
Author(s) -
Fradkin Amber Haynes,
Boand Carl S.,
Eisenberg Stephen P.,
Rosendahl Mary S.,
Randolph Theodore W.
Publication year - 2010
Publication title -
biotechnology progress
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.572
H-Index - 129
eISSN - 1520-6033
pISSN - 8756-7938
DOI - 10.1002/btpr.393
Subject(s) - inclusion bodies , immunogenicity , recombinant dna , circular dichroism , solubility , hydrostatic pressure , escherichia coli , solubilization , chemistry , biochemistry , monomer , chromatography , high pressure , microbiology and biotechnology , biology , antibody , polymer , organic chemistry , gene , physics , immunology , thermodynamics , engineering physics , engineering
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. © 2010 American Institute of Chemical Engineers Biotechnol. Prog., 2010