Lizozim Saflaştırılması İçin Heparin İmmobilize Edilmiş poli(2-hidroksi etilmetakrilat) Kriyojeller

Yumurta akından lizozim saflaştırılması için heparin immobilize edilmiş poli(hidroksi etilmetakrilat) (PHEMA) kriyojeller sentezlenmiştir. İlk olarak, PHEMA kriyojeller kriyopolimerizasyon yöntemi ile sentezlenmiştir. Heparin, siyanojen bromür aktivasyonu ile PHEMA kriyojeline kovalent olarak immobilize edilmiştir. PHEMA kriyojel yapısının heparin ile modifikasyonu Fourier dönüşümlü kızıl ötesi spektroskopisi (FTIR) ile incelenmiştir. PHEMA kriyojellerin yüzey ve iç yapı morfolojileri taramalı elektron mikroskobu (SEM) ile karakterize edilmiştir. PHEMA kriyojelin yüzey alanı 25.2 m2 /g olarak bulunmuştur. Heparin immobilize edilmiş PHEMA kriyojeller lizozim adsorpsiyon çalışmalarında pH, lizozim derişimi, akış hızı, sıcaklık ve iyonik şiddetin etkilerini incelemek için kullanılmıştır. Optimum koşullar altında, heparin immobilize edilmiş PHEMA kriyojelin sulu çözeltide en yüksek lizozim adsorpsiyonu 48.73 mg/g olarak bulunmuştur. Sürekli sistemde lizozimin desorpsiyonu için 1.0 M NaCI çözeltisi kullanılmıştır. Heparin immobilizasyonuna heparin derişiminin etkisi incelenmiştir. Heparin immobilize edilmiş PHEMA kriyojellerin tekrar kullanılabilirliği adsorpsiyon-desorpsiyon döngüsü 10 kez tekrarlanarak test edilmiştir. Langmuir adsorpsiyon modeli çizilmiş ve adsorpsiyon çalışmaları için uygun bulunmuştur. Yumurta akından lizozim saflığı %12 ayırıcı jel kullanılarak sodyum-dodesil sülfat poliakrilamid jel elektroforezi kullanılarak analiz edilmiştir.

Biologically Modified poly(2-hydroxylethyl methacrylate) Cryogels for Lysozyme Purification

Heparin immobilized poly(2-hydroxylethyl methacrylate) PHEMA cryogel was synthesized and applied for lysozymepurification from egg white. Firstly, the PHEMA cryogel was synthesized by cryopolymerization and then heparin wascovalently immobilized on to the PHEMA cryogel with cyanogen bromide activation. The modification of PHEMA cryogelstructure with heparin was further confirmed by Fourier-transform infrared spectroscopy (FTIR). The surface and innerstructure morphologies of PHEMA cryogels were studied and characterized by the scanning electron microscope (SEM). Thesurface area of PHEMA cryogel was found to be 25.2 m2/g. Heparin immobilized PHEMA cryogels were used in lysozymeadsorption studies to assess the effects of pH, lysozyme concentration, flow rate, temperature and ionic strength. Themaximum lysozyme adsorption on the heparin immobilized PHEMA cryogel was found to be 48.73 mg/g from aqueoussolutions under optimized conditions. 1.0 M NaCI solution was used for desorption of lysozyme in a continuous system.The reusability of heparin immobilized PHEMA cryogels was tested for 10 adsorption-desorption cycles. The Langmuiradsorption model was plotted and found fitted for adsorption studies. The purity of lysozyme from egg white studies wasanalysed by sodium-dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) using 12% separating gel.

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