Iranian Biomedical Journal
مجله بیومدیکال ایران
IBJ
Basic Sciences
http://ibj.pasteur.ac.ir
1
admin
1028-852X
2008-823X
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10.61186/ibj
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8888
-
en
jalali
1394
1
1
gregorian
2015
4
1
19
2
online
1
fulltext
en
توسعه فرایند کارآمد تولید GCSF نوترکیب انسانی در اشرشیا کلی
Efficient Process Development of Recombinant Human Granulocyte Colony-Stimulating Factor (rh-GCSF) Production in Escherichia coli
Related Fields
Related Fields
مقاله کامل
Full Length/Original Article
Background: The protein hormone granulocyte colony-stimulating factor (GCSF) stimulates the production of white blood cells and plays an important role in medical treatment of cancer patients. Methods: An efficient process was developed for heterologous expression of human GCSF in E. coli BL21 (DE3). The feeding rate was adjusted to achieve the maximum attainable specific growth rate under critical value. In this method, specific growth rate was maintained at the maximum value of 0.55 h-1 at the beginning of feeding to 0.4 h-1 at the induction time. Recombinant human GCSF (rh-GCSF) was produced as inclusion body. At first, inclusion bodies were released by cell disruption and then washed, solubilized and refolded. Finally, the rh-GCSF was purified by cation exchange chromatography. Results: Obviouly, higher specific growth rate decreases process time and consequently increases productivity. The final concentration of biomass and GCSF was achieved 126 g DCW.l-1 and 32.1 g.l-1. Also, the final specific yield (YP/X) and total productivity of rh-GCSF were obtained 254 mg.g-1 DCW and 1.83 g.l-1.h-1, respectively. According to the available data, this is one of the highest YP/X and productivity that has been reported for any human protein which is expressed in E. coli. Recovery yield of purification process was %40 and purity of recombinant protein was over than 99%. The circular dichroism spectra of purified rh-GCSF, Neupogen® and PD-Grastim showed that all proteins have a similar secondary structure. Conclusion: Modified exponential feeding strategy for fed-batch cultivation of recombinant E. coli, results in minimum fed-batch duration and maximum productivity.
Escherichia coli, Granulocyte colony-stimulating factor (GCSF), Process development
102
110
http://ibj.pasteur.ac.ir/browse.php?a_code=A-10-1-546&slc_lang=en&sid=1
Valiollah
Babaeipour
ولی الله
بابایی پور
babaeipour@mu.ac.ir
100319475328460055973
100319475328460055973
Yes
Sirwan
Khanchezar
سیروان
خوانچه زر
100319475328460055974
100319475328460055974
No
Mohammad Reza
Mofid
محمدرضا
مفید
100319475328460055975
100319475328460055975
No
Mahdi
Pesaran Hagi Abbas
مهدی
پسران حاجی عباس
100319475328460055976
100319475328460055976
No