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040			_aAIKTC-KRRC _cAIKTC-KRRC
100			_97240 _aEslami Samarin, Z.
245			_aResponse Surface Optimization of the Expression Conditions for Synthetic Human Interferon alpha-2b Gene in Escherichia coli
250			_aVol. 80(03), September-October
260			_aMumbai _c2018 _bIndian Journal of Pharmaceutical Science
300			_a470-479
520			_aRecombinant human interferon alpha-2b is an FDA-approved drug for monotherapy or in combination therapy with other drugs for hepatitis and cancers. It belongs to a family of homologous proteins involved in antiviral, antiproliferative, and immunoregulatory processes. A different expression system has been used for overexpression of this protein. Escherichia coli expression system is a highly characterized host and various expression settings have been developed based on its properties. However, finding the best conditions for the overexpression of recombinant human interferon alpha-2b remains to be addressed. In this study, the expression of synthetic human interferon alpha-2b gene in E. coli was greatly improved by adjusting the expression condition. In this regard, a recombinant gene was designed and codon optimized for the periplasmic expression of this protein. Then, gene subcloning was employed to insert the synthesized gene into the pET22b expression vector. Thereafter, the response surface methodology method was employed to design 20 experiments to find out the optimum points for isopropyl β-D-1-thiogalactopyranoside concentration, post-induction period, and the cell density of induction (OD600). The expression fluctuations were assessed by using the real time polymerase chain reaction method. Our results indicated that the synthetic human interferon alpha-2b gene was successfully codon optimized and subcloned into the expression vector. The real time polymerase chain reaction results revealed that the optimum levels of the selected parameters are 0.27 mM for isopropyl β-D-1-thiogalactopyranoside concentration, 7.98 H for the post-induction period, and 3.93 for cell density (OD600). These optimized conditions led to a 3.5-fold increase in the rhIFNα2b expression, which is highly promising for large scale rhIFNα2b overexpression.
650		0	_94639 _aPHARMACEUTICS
653			_aCodon optimization
653			_aResponse surface methodology
700			_97241 _a Abolghasem, S.
700			_97242 _aDehnav, E.
773	0		_tIndian journal of pharmaceutical sciences _x0250-474X _dNew Delhi Indian Pharmaceutical Association
856			_yClick here _uhttp://www.ijpsonline.com/articles/response-surface-optimization-of-the-expression-conditions-for-synthetic-human-interferon-alpha2b-gene-in-iescherichia-colii-3488.html
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