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    • ISSN: 2010-0264
    • Frequency: Bimonthly (2010-2014); Monthly (Since 2015)
    • DOI: 10.18178/IJESD
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Editor-in-chief
The University of Queensland, Australia
It is my honor to be the editor-in-chief of IJESD. The journal publishes good papers in the field of environmental science and development.
IJESD 2010 Vol.1(2): 111-114 ISSN: 2010-0264
DOI: 10.7763/IJESD.2010.V1.22

An efficient purification method for high recovery of Recombinant Human Granulocyte Colony Stimulating Factor from recombinant E. coli

S. Abolghasemi Dehaghani, V. Babaeipour, M. R. Mofid, A. Divsalar, F. Faraji

Abstract—Human G-CSF, a single chain polypeptide containing 174 amino acid residues (MW=18,800, pI=6.1), is one of the hemopoietic growth factors. Development of inexpensive and simple culture media is always favorable for commercial production of recombinant proteins in E. coli. The high-level expression of eukaryotic proteins in E. coli often leads to formation of insoluble inclusion bodies (IBs) in the cytoplasm or periplasm. Recovery of active material from (IBs) is often difficult and involves two general steps: protein solubilization in a denaturant and protein refolding. On a commercial scale, reducing the number of protein purification steps is practical and economical because each purification step not only increases the final product but also causes successive yield losses of the recombinant protein. In this research, we developed an efficient and scalable procedure for production and purification of recombinant human (rh-GCSF) of E. coli. This process include: an optimized batch culture with LB and glucose 10 g/l with expression level 40%, cell harvesting, cell lyses with high pressure homogenizer, two steps washing, IB solubilization, refolding, and finally protein purified by FPLC with cation exchanger column. By the using of the new developed method, of 1.8 g l-1 rh-GCSF was produced in each batch, 720 mg pure of recombinant protein was obtained with recovery yield about 40% and purity over than 99%. According to available data this is one of the highest yield and production level of the purified recombinant protein that has been reported for human recombinant protein which is expressed in E. coli. Also with this procedure we can produce a protein that the structural characterization was preserved.

Index Terms—rh-GCSF; Escherichia coli; Inclusion bodies; purification; Solubilization; Refolding

S. Abolghasemi Dehaghani is with the Dept. Biology‚ Science & Research Branch‚ Islamic Azad University, Tehran, Iran, (corresponding author to provide phone; fax: +98+21-88970258; e-mail: bio86.shai@yahoo.com ).
V. Babaeipour is with the Biochemical Engineering Group, Biotechnology Research Center, Tehran, Iran,(corresponding author to provide phone; fax: +98+21-22974605; e-mail: baba1@sbmu.ac.ir).
M. R. Mofid is with the Isfahan University of medical sciences (e-mail: mohrezamofid@hotmail.com, mofid @pharm.mui.ac.ir).
A. Divsalar is with the Department of Biological Sciences, Tarbiat Moallem University, Tehran, Iran,( e-mail: divsalar@ibb.ut.ac.ir )
F. Faraji is with the Dept. Biology‚ Science & Research Branch‚ Islamic Azad University, Tehran, Iran,( e-mail: visionbsh@gmail.com).

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Cite: S. Abolghasemi Dehaghani, V. Babaeipour, M. R. Mofid, A. Divsalar, F. Faraji, "An efficient purification method for high recovery of Recombinant Human Granulocyte Colony Stimulating Factor from recombinant E. coli," International Journal of Environmental Science and Development vol. 1, no. 2, pp. 111-114, 2010.

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