Abstract
The bacterial cellulose (BC) nanofibers were produced by Acetobacter xylinum in the Hestrin-Schramm medium in a static condition for 14 days. Then, Cibacron Blue F3GA (CB) was covalently attached onto the BC nanofibers for affinity adsorption of recombinant interferon-α (rHuIFN-α). The CB content of the BC nanofibers was 178 μmol/g. The specific surface area of the BC nanofibers was determined to be 914 m2/g. rHuIFN-α adsorption studies were performed by batch system. The non-specific adsorption of rHuIFN-α on the BC nanofibers was very low (2.1 mg/g polymer). CB attachment onto the BC nanofibers significantly increased the rHuIFN-α adsorption (1620 mg/g). The maximum rHuIFN-α adsorption was observed at pH 6.0. The rHuIFN-α adsorption capacity decreased drastically with an increase of the aqueous phase concentration of sodium chloride. The elution studies were performed by adding 1 M NaCl to the rHuIFN-α solutions in which adsorption equilibria had been reached. The eluted rHuIFN-α had a specific activity in the range of of 3.32-3.45 x 108 IU/mg as inhibition of the cytopathic effect of MDBK cells. In order to determine the effects of adsorption conditions on possible conformational changes of rHuIFN-α structure, fluorescence spectrophotometry was employed. We resulted that the BC-CB nanofibers can be applied for rHuIFN-α adsorption without causing any significant conformational changes. The elution results showed that the binding of rHuIFN-α to the BC-CB nanofibers was reversible. This study demonstrated that the BC nanofibers has a potential for affinity carrier applications in protein adsorption systems.
| Original language | English |
|---|---|
| Title of host publication | Protein Purification |
| Publisher | Nova Science Publishers, Inc. |
| Pages | 185-197 |
| Number of pages | 13 |
| ISBN (Print) | 9781614700982 |
| Publication status | Published - Feb 2012 |
Keywords
- Dye affinity carriers
- Interferon
- Nanofibers
- Protein purification
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