S. Senel et al., Zinc ion-promoted adsorption of lysozyme to Cibacron Blue F3GA-attached microporous polyamide hollow-fiber membranes, COLL SURF A, 182(1-3), 2001, pp. 161-173
Citations number
38
Categorie Soggetti
Physical Chemistry/Chemical Physics
Journal title
COLLOIDS AND SURFACES A-PHYSICOCHEMICAL AND ENGINEERING ASPECTS
Dye-affinity and metal chelate affinity adsorption are increasingly used fo
r protein separation. Synthetic hollow fiber membranes have advantages as s
upport matrices in comparison to conventional bead supports because they ar
e not compressible and they eliminate internal diffusion limitations. The g
oal of this study was to explore in detail the performance of hollow fibers
composed of modified polyamide to which Cibacron Blue F3GA and Zn(II) were
attached for adsorption of lysozyme. The polymer matrix was characterized
by scanning electron microscopy. These dye-affinity and Zn(II) chelated hol
low-fibers were used in the lysozyme adsorption-elution studies. The effect
s of initial concentration of lysozyme and medium pH on the adsorption effi
ciency of dye-attached and metal-chelated hollow-fibers were studied in a b
atch reactor. The effect of Zn(II) loading on lysozyme adsorption was also
studied. The non-specific adsorption of lysozyme on the polyamide hollow-fi
bers was 1.8 mg g(-1). Cibacron Blue F3GA attachment significantly increase
d the lysozyme adsorption up to 63.2 mg g(-1). Lysozyme adsorption capacity
of the Zn(II) chelated hollow-fibers (144.2 mg g(-1)) was greater than tha
t of the Cibacron Blue F3GA-attached hollow-fibers. A significant amount of
the adsorbed lysozyme (up to 97%) was eluted in 1 h in the elution medium
containing 1.0 M NaSCN at pn 8.0 and 25 mM EDTA at pH 4.9. In order to exam
ine the effects of separation conditions on possible conformational changes
of lysozyme structure, fluorescence spectrophotometry was employed. We con
clude that dye- and metal-chelate affinity chromatography with polyamide ho
llow-fibers can be applied for lysozyme adsorption without causing any sign
ificant conformational changes and denaturation. Repeated adsorption/elutio
n processes showed that these novel dye-attached and Zn(II) chelated hollow
-fibers are suitable for lysozyme adsorption. (C) 2001 Elsevier Science B.V
. All rights reserved.