Single-step purification of recombinant green fluorescent protein on expanded beds of immobilized metal affinity chromatography media
| Autor: | Sohel Dalal, Smita Raghava, Munishwar N. Gupta |
|---|---|
| Rok vydání: | 2008 |
| Předmět: |
inorganic chemicals
Packed bed Environmental Engineering Chromatography Chemistry Biomedical Engineering Langmuir adsorption model Bioengineering medicine.disease_cause Green fluorescent protein law.invention symbols.namesake Adsorption Column chromatography Affinity chromatography law cardiovascular system symbols medicine Recombinant DNA tissues Escherichia coli Biotechnology |
| Zdroj: | Biochemical Engineering Journal. 42:301-307 |
| ISSN: | 1369-703X |
| DOI: | 10.1016/j.bej.2008.07.010 |
| Popis: | Immobilized metal ion affinity chromatography (IMAC) in expanded bed mode is used for purifying recombinant green fluorescent protein (GFP) overexpressed in Escherichia coli. The purification is carried out on two different matrices, i.e. Ni2+ Streamline™ and Ni2+ cross-linked alginate beads. The binding isotherms to both IMAC media followed the Langmuir model. The maximum binding capacity (qmax) of Ni2+ Streamline™ and Ni2+ cross-linked alginate for the GFP was 1,42,860 FU ml−1 and 18,000 FU ml−1, respectively. The expanded bed column chromatography using Ni2+ Streamline™ gave 2.7-fold purification with 89% of GFP recovery, while Ni2+ alginate gave 3.1-fold purification with 91% of GFP recovery. SDS-PAGE of purified GFP in both cases showed single band. The results obtained in the expanded bed chromatography are compared with those obtained in packed bed chromatography. |
| Databáze: | OpenAIRE |
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