title: Optimizing conditions for production of high levels of soluble recombinant human growth hormone using Taguchi method.
abstract: Human growth hormone (hGH) is synthesized and stored by somatotroph cells of the anterior pituitary gland and can effect on body metabolism. This protein can be used to treat hGH deficiency, Prader-Willi syndrome and Turner syndrome. The limitations in current technology for soluble recombinant protein production, such as inclusion body formation, decrease its usage for therapeutic purposes. To achieve high levels of soluble form of recombinant human growth hormone (rhGH) we used suitable host strain, appropriate induction temperature, induction time and culture media composition. For this purpose, 32 experiments were designed using Taguchi method and the levels of produced proteins in all 32 experiments were evaluated primarily by ELISA and dot blotting and finally the purified rhGH protein products assessed by SDS-PAGE and Western blotting techniques. Our results indicate that media, bacterial strains, temperature and induction time have significant effects on the production of rhGH. The low cultivation temperature of 25°C, TB media (with 3% ethanol and 0.6M glycerol), Origami strain and a 10-h induction time increased the solubility of human growth hormone.
authors.affiliation: University of Isfahan, Isfahan, Iran. Electronic address: Marzieh.savari@gmail.com.
authors.forename: Marzieh
authors.initials: M
authors.lastname: Savari
authors.affiliation: Department of Biology, Faculty of Sciences, University of Isfahan, Isfahan 81746-73695, Iran. Electronic address: s.h.zarkesh@sheffield.ac.uk.
authors.forename: Sayyed Hamid
authors.initials: SH
authors.lastname: Zarkesh Esfahani
authors.affiliation: Department of Pathology, Medical School, Isfahan University of Medical Sciences, Hezar Jerib Street, Isfahan 81746-73695, Iran. Electronic address: masoud_edalati@hotmail.com.
authors.forename: Masoud
authors.initials: M
authors.lastname: Edalati
authors.affiliation: Department of Biotechnology, Faculty of Advanced Sciences and Technologies, University of Isfahan, Isfahan, Iran. Electronic address: d.biria@ast.ui.ac.ir.
authors.forename: Davoud
authors.initials: D
authors.lastname: Biria
mesh_terms: D004797:Enzyme-Linked Immunosorbent Assay; D004926:Escherichia coli / Q000235:genetics*; D019382:Human Growth Hormone / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism*; D006801:Humans; D011994:Recombinant Proteins / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism*; D012107:Research Design; D013696:Temperature
chemical_list: D011994:Recombinant Proteins; D019382:Human Growth Hormone
keywords: Escherichia coli; Human growth hormone; Soluble protein; Taguchi software