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:: Volume 3, Number 2 (Autumn-winter 2015-2016 2015) ::
Hakim Jorjani J 2015, 3(2): 89-96 Back to browse issues page
Characterization of recombinant Granulocyte Colony Stimulating Factor (G-CSF) protein by Extinction Coefficient Calculation
Fariba Bagherieh
M.Sc. of Medical Biotechnology, Faculty of Advanced Medical Technologies, Golestan University of Medical Sciences, Gorgan, Iran
Abstract:   (1606 Views)

Background and Objective: Extinction coefficient is the unique value to each protein and can evaluate the type and purity of recombinant protein produced in laboratories and factories producing recombinant proteins. In this study, a simple method for calculating the extinction coefficient of recombinant protein Granulocyte-Colony Stimulating Factor (G-CSF) is presented.

Methods: G-CSF with concentration of 0.5 mg/ml was prepared from Arya Tina Gene. Sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) method was used to check the purity of the protein, then serial dilutions of 1/2 to 1/64 of the protein were prepared and using UV-VIS spectroscopy the optical density was measured.

Results: Electrophoresis result as a single band of 18 kDa was observed. The results of UV-VIS spectroscopy of serial dilution were plotted as a distinct spectrograms and these were used to plot linear regression curve of the standard concentration of G-CSF.

Conclusion: The results of this study shows the extinction coefficient equal to 0.81 M-1 cm-1 for recombinant G-CSF that was accordant with the number reports by international producers of G-CSF and suggests the correct method is used to calculate the physicochemical parameter.

Keywords: Extinction coefficient (), UV-VIS Spectroscopy, Recombinant Granulocyte-Colony stimulating Factor (G-CSF), sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE)
Full-Text [PDF 795 kb]   (691 Downloads)    
Type of Study: Original paper | Subject: General
Received: 2016/03/19 | Accepted: 2016/03/19 | Published: 2016/03/19
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Bagherieh F. Characterization of recombinant Granulocyte Colony Stimulating Factor (G-CSF) protein by Extinction Coefficient Calculation. Hakim Jorjani J. 2015; 3 (2) :89-96
URL: http://goums.ac.ir/jorjanijournal/article-1-410-en.html
Volume 3, Number 2 (Autumn-winter 2015-2016 2015) Back to browse issues page
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