Cu2+ - takılı sporopollenin gömülü süpermakrogözenekli monolitik phema kriyojel afinite membranlar ile sitokrom c adsorpsiyonu
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Tarih
2012
Yazarlar
Dergi Başlığı
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Cilt Başlığı
Yayıncı
Aksaray Üniversitesi Fen Bilimleri Enstitüsü
Erişim Hakkı
info:eu-repo/semantics/openAccess
Özet
Bu çalışmada Cu2+ - takılı sporopollenin gömülü süpermakrogözenekli monolitik PHEMA kriyojel afinite membranlar ile sitokrom c adsorpsiyonu incelenmiştir. Bu amaçla, ilk olarak 20 ? m çapındaki sporopollenin partikülleri, 150 ppm Cu2+ çözeltisi (pH 5) ile muamele edilmiştir. Daha sonra, 15 mg Cu2+- takılı sporopollenin gömülü PHEMA bazlı monolitik kriyojeller, monomer olarak 2-hidroksietil metakrilat (HEMA) ve çapraz bağlayıcı olarak N,N'-metilen-bis-akrilamid'in (MBAAm) radikal kriyo-kopolimerizasyonu yöntemiyle 5 mL'lik plastik şırıngalarda sentezlenmiştir. Cu2+-takılı sporopollenin gömülü kriyojel afinite sorbentler farklı koşullarda (pH, başlangıç sitokrom c konsantrasyonu, sıcaklık, akış hızı ve iyonik şiddet) adsorpsiyon çalışmalarında kullanılmıştır. Hazırlanan kriyojeller, şişme testleri ve taramalı elektron mikroskopu (SEM) ile karakterize edilmiştir. Sulu çözeltiden maksimum sitokrom c adsorpsiyonu pH 7,0'de gözlenmiştir. Yapılan derişim taramasında yaklaşık 3,0 mg/mL konsantrasyonda dengeye ulaşılmıştır. Ayrıca, sitokrom c adsorpsiyonunun artan akış hızı ve sıcaklık ile ters orantılı olduğu gözlenmiştir. Cu2+-takılı olmayan sporopollenin gömülü kriyojel afinite sorbentlere non-spesifik sitokrom c adsorpsiyonu ihmal edilebilir düzeydedir. Kriyojel afinite sorbentlere, adsorpsiyon kapasitesinde önemli bir azalma olmaksızın 30 defa adsorpsiyon/desorpsiyon işlemi uygulanmıştır.
In this study, adsorption of cytochrome c onto supermacroporous monolithic PHEMA affinity cryogels embedded with Cu2+ - attached sporopollenin was investigated. For this purpose, firstly, sporopollenin particles, with size of 20 ? m, were treated with 150 ppm Cu2+ solution at pH 5,0 to bond Cu2+ ions to the structure. Then, PHEMA based monolithic affinity cryogels containing 15 mg of embedded Cu2+- sporopollenin were synthesized by free radical polymerization using, 2-hydroxyethyl methacrylate (HEMA) as a monomer and N,N´-methylene-bis-acrylamide (MBAAm) as a cross-linker in a plastic syringe of 5 mL. Monolithic affinity cryogel sorbents embedded with Cu2+- sporopollenin were used in adsorption studies by changing the conditions (pH, initial cytochrome c concentration, temperature, flow rate and ionic strength). The prepared cryogels were characterized by swelling tests and scanning electron microscopy (SEM). The maximum amount of cytochrome c adsorption from aqueous solution was observed at pH 7,0. In concentration studies, the equilibrium was established about 3,0 mg/mL of cytochrome c concentration. In addition, it was observed that cytochrome c adsorption was inversely proportional to flow rate and temperature. It was also observed that non-specific adsorption of cytochrome c onto affinity cryogel membranes containing sporopollenin particles, without atteched Cu2+, was ignoreable degree. Adsorption/desorption processes were applied 30 times with these affinity cryogel sorbents without noticeable loss in their adsorption capacity.
In this study, adsorption of cytochrome c onto supermacroporous monolithic PHEMA affinity cryogels embedded with Cu2+ - attached sporopollenin was investigated. For this purpose, firstly, sporopollenin particles, with size of 20 ? m, were treated with 150 ppm Cu2+ solution at pH 5,0 to bond Cu2+ ions to the structure. Then, PHEMA based monolithic affinity cryogels containing 15 mg of embedded Cu2+- sporopollenin were synthesized by free radical polymerization using, 2-hydroxyethyl methacrylate (HEMA) as a monomer and N,N´-methylene-bis-acrylamide (MBAAm) as a cross-linker in a plastic syringe of 5 mL. Monolithic affinity cryogel sorbents embedded with Cu2+- sporopollenin were used in adsorption studies by changing the conditions (pH, initial cytochrome c concentration, temperature, flow rate and ionic strength). The prepared cryogels were characterized by swelling tests and scanning electron microscopy (SEM). The maximum amount of cytochrome c adsorption from aqueous solution was observed at pH 7,0. In concentration studies, the equilibrium was established about 3,0 mg/mL of cytochrome c concentration. In addition, it was observed that cytochrome c adsorption was inversely proportional to flow rate and temperature. It was also observed that non-specific adsorption of cytochrome c onto affinity cryogel membranes containing sporopollenin particles, without atteched Cu2+, was ignoreable degree. Adsorption/desorption processes were applied 30 times with these affinity cryogel sorbents without noticeable loss in their adsorption capacity.
Açıklama
Anahtar Kelimeler
Kriyojel, PHEMA, Sporopollenin, Sitokrom c, İMAK, Adsorpsiyon, Cryogel, Cytochrome c, IMAC, Adsorption
