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Indian Journal of Biochemistry and Biophysics (IJBB) >
IJBB Vol.49 [2012] >
IJBB Vol.49(4) [August 2012] >
| Title: | Refolding of recombinant human granulocyte colony stimulating factor: Effect of cysteine/cystine redox system |
| Authors: | Tiwari, Krishnanand
Shebannavar, Sunil
Kattavarapu, Krishna
Pokalwar, Santosh
Mishra, Maheshwari K
Chauhan, Ugam Kumari |
| Keywords: | Granulocyte colony-stimulating factor
Refolding
Redox system
Cysteine/cystine
Oxidative refolding |
| Issue Date: | Aug-2012 |
| Publisher: | NISCAIR-CSIR, India |
| Abstract: | Granulocyte
colony-stimulating factor (G-CSF) is a multi-functional cytokine which is
widely used for treating neutropenia in humans. Evaluation of alternative to
expensive components of redox buffer (reduced and oxidized glutathione) is an
important step in reducing the cost of production of human biotherapeutic proteins.
In the present study, refolding of recombinant human
G-CSF expressed as inclusion bodies (IBs) in E. coli was optimized using cysteine and cystine redox agents. The
refolding to correct native form of G-CSF was assessed by reverse
phase high performance liquid chromatography (RP-HPLC). The optimized
concentrations of cysteine and cystine for correct refolding of G-CSF were
found to be 2 mM and 1 mM, respectively. The correctly refolded G-CSF was
detected as early as 4 h of incubation in renaturation buffer containing
optimized concentrations of cysteine (2 mM) and cystine (1 mM) redox agents.
Refolding of G-CSF in optimized redox system increased with increase in shuffling
time. Overall, the results suggested the use of cysteine/cystine redox pair could
be an alternative to the costlier redox pairs for successful refolding of G-CSF
and possibly other human biotherapeutic proteins of importance. |
| Page(s): | 285-288 |
| CC License: |  CC Attribution-Noncommercial-No Derivative Works 2.5 India |
| ISSN: | 0975-0959 (Online); 0301-1208 (Print) |
| Source: | IJBB Vol.49(4) [August 2012]
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