Affinity adsorption of recombinant human interferon-alpha on a porous dye-affinity adsorbent


Karakoc V., Yavuz H. , Denizli A.

COLLOIDS AND SURFACES A-PHYSICOCHEMICAL AND ENGINEERING ASPECTS, cilt.240, ss.93-99, 2004 (SCI İndekslerine Giren Dergi) identifier identifier

  • Cilt numarası: 240
  • Basım Tarihi: 2004
  • Doi Numarası: 10.1016/j.colsurfa.2004.03.005
  • Dergi Adı: COLLOIDS AND SURFACES A-PHYSICOCHEMICAL AND ENGINEERING ASPECTS
  • Sayfa Sayıları: ss.93-99

Özet

Interferons are potent biologically active proteins synthesized and secreted by somatic cells of all mammalian species. Dye-affinity adsorption is increasingly used for protein separation. Hollow fibers have several advantages as adsorbents in comparison to conventional bead supports because they are not compressible and they eliminate internal diffusion limitations. The aim of this study was to explore in detail the performance of polyamide hollow fibers to which Cibacron Blue F3GA was attached for adsorption of recombinant interferon-alpha (rHuIFN-alpha). The hollow fiber was characterized by scanning electron microscopy. These dye-carrying hollow fibers (35.8 mumol/g) were used in the rHuIFN-alpha adsorption-elution studies. The effects of initial concentration of rHuIFN-alpha, medium pH, ionic strength and temperature on the adsorption efficiency of dye-attached hollow fibers were studied in a batch system. The non-specific adsorption of rHuIFN-alpha on the hollow fibers was 1.2 mg/g. Cibacron Blue F3GA attachment significantly increased the rHuIFN-alpha adsorption up to 99.8 mg/g. Significant amount of the adsorbed rHuIFN-alpha (up to 94.8%) was eluted in 1 h in the elution medium containing 1.0 M NaCl. In order to determine the effects of adsorption conditions on possible conformational changes of rHuIFN-alpha structure, fluorescence spectrophotometry was employed. We resulted that dye-affinity hollow fibers can be applied for rHuIFN-alpha adsorption without causing any significant conformational changes. Repeated adsorption/elution processes showed that these dye-attached hollow fibers are suitable for rHuIFN-alpha adsorption. (C) 2004 Elsevier B.V. All rights reserved.