| Autor: |
Esfandiar S; Biotechnology Group, Chemical Engineering Department, Tarbiat Modares University, P.O. Box 14115-143, Tehran, Iran., Hashemi-Najafabadi S, Shojaosadati SA, Sarrafzadeh SA, Pourpak Z |
| Jazyk: |
angličtina |
| Zdroj: |
Biotechnology and applied biochemistry [Biotechnol Appl Biochem] 2010 Apr 14; Vol. 55 (4), pp. 209-14. Date of Electronic Publication: 2010 Apr 14. |
| DOI: |
10.1042/BA20090256 |
| Abstrakt: |
The expression of rhIL-2 (recombinant human interleukin-2) in bacteria results in the formation of insoluble inclusion-body aggregates. These aggregates were first solubilized under denaturing conditions (sodium phosphate buffer solution containing 8 M urea and 10 mM 2-mercaptoethanol) and then purified using IMAC (immobilized metal-ion-affinity chromatography). IMAC was used to capture rhIL-2. The protein was gradually refolded on the column by a gradient elution (8 M to 0 M urea) in the presence of 10% (v/v) glycerol. Glycerol was used to prevent protein aggregation during the refolding step. Using this method, rhIL-2 was collected at 97% purity and its activity was measured by the lymphocyte transformation test. The measured activity was identical with commercial human interleukin-2. |
| Databáze: |
MEDLINE |
| Externí odkaz: |
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