Protein A-immobilized microporous polyhydroxyethylmethacrylate affinity membranes for selective sorption of human-immunoglobulin-G from human plasma
Yükleniyor...
Tarih
2000
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Vsp Bv
Erişim Hakkı
info:eu-repo/semantics/closedAccess
Özet
Microporous membranes made of poly(2-hydroxyethylmethacrylate) [poly(HEMA)] carrying protein A were used for selective sorption of human-IgG from human plasma. Poly(HEMA) membranes were prepared by a photo-polymerization technique, and activated by cyanogen bromide (CNBr) in an alkaline medium (pH 11.5). Bioligand protein A was then immobilized by covalent binding onto these CNBr-activated membranes. The amount of immobilized protein A was controlled by changing pH and the initial concentrations of CNBr and protein A. The non-specific adsorption of protein A on the plain poly(HEMA) membranes was 2.9 mu g cm(-2). Maximum protein A immobilization was observed at pH 9.5. Up to 180 mu g cm(-2) was immobilized on the CNBr-activated poly(HEMA) membranes. The maximum adsorption of human-IgG on the protein A-immobilized poly(HEMA) membranes was observed at pH 8.0. The non-specific adsorption of human-IgG onto the plain poly(HEMA) membranes was low (about 4.4 mu g cm(-2)). Higher human-IgG adsorption values (up to 394 mu g cm(-2)) were obtained in which the protein A-immobilized poly(HEMA) membranes were used. Much higher amounts of human-Igg (up to 489 mu g cm(-2)) were adsorbed from human plasma. Up to 91 % of the adsorbed human-IgG was desorbed by using 0.1 M aminoacetic acid as elution agent. The adsorption-desorption cycle was repeated ten times using the same polymeric membranes. There was no remarkable reduction in the adsorption capacity of the protein A-immobilized poly(HEMA) membranes.
Açıklama
Anahtar Kelimeler
Kaynak
Journal Of Biomaterials Science-Polymer Edition
WoS Q Değeri
Q1
Scopus Q Değeri
Q1
Cilt
11
Sayı
4
Künye
closedAccess
