Characterization of recombinant Granulocyte Colony Stimulating Factor (G-CSF) protein by Extinction Coefficient Calculation

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 ۰.۵ 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 ۱/۲ to ۱/۶۴ of the protein were prepared and using UV-VIS spectroscopy the optical density was measured. Results: Electrophoresis result as a single band of ۱۸ 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 ۰.۸۱ M-۱ cm-۱ 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) , ضریب خاموشی , طیف سنجی فرابنفش - مرئی , پروتئین نوترکیب فاکتور محرک گرانولوسیتی (G-CSF) , الکتروفورز در ژل پلی-آکریل آمید سدیم دودسیل سولفات (SDS-PAGE)