Application of Cu2+-attached Magnetite Nanoparticles Embedded Supermacroporous Monolithic Composite Cryogels for DNA Adsorption

Abstract

Cryogels are known as a novel generation of stationary phases in the separation science, and they have been succesfully used as efficient adsorbents for separation and purification of biomolecules, such as proteins, enzymes, DNA, cell organelles and viruses in downstream processes. In this study, DNA adsorption performance of Cu2+-attached magnetite Fe3O4 nanoparticles Cu2+-AMNPs embedded supermacroporous monolithic composite cryogels were investigated. Firstly, Magnetite nanoparticles NPs coated with 3-aminopropyltriethoxysilane APTES were prepared by silanization reaction. Then, Cu2+ ions were attached to silanized magnetite NPs through amine groups. After that, Cu2+-AMNPs were embedded into the cryogels for DNA adsorption studies. The prepared cryogel was characterized by scanning electron microscopy SEM , elemental analysis, FTIR and swelling. SEM analysis indicates that the cryogel have a heteroporous structure with interconnected pores of 10-50 m size, which ascribed to the porogens effect of frozen water crystals. The maximum amount of DNA adsorption was 19.97 mg/g polymer, at pH 8.0.

Keywords

• Affinity chromatography,DNA adsorption,metal chelate,cryogels

Cu2+-takılı Magnetit Nanopartikül Gömülü Süpermakrogözenekli Monolitik Kompozit Kriyojellerin DNA Adsorpsiyon Uygulaması

Abstract

Kriyojeller ayırma biliminde yeni nesil sabit faz olarak değerlendirilmektedirler. Kriyojeller, altakım işlemlerinde protein, enzim, DNA, hücre organelleri ve virüsler gibi biyomoleküllerin ayrılması ve saflaştırılmasında etkin adsorbentler olarak kullanılmaktadır. Bu çalışmada, Cu2+-takılı magnetit Fe3O4 nanopartikül Cu2+-AMNP gömülü süpermakrogözenekli monolitik kompozit kriyojellerin DNA adsorpsiyon performansı incelenmiştir. Öncelikle, 3-aminopropyltriethoxysilane APTES ile kaplanmış magnetit NP’ler silanizasyon reaksiyonu ile hazırlanmıştır. Daha sonra, silanlanmış magnetit NP’lere, amin grupları üzerinden Cu2+ iyonları takılmıştır. Bundan sonra, Cu2+-AMNP’ler DNA adsorpsiyon çalışmaları için süpermakrogözenekli monolitik kompozit kriyojellere gömülmüştür. Hazırlanan kriyojel taramalı elektron mikroskobu SEM , elementel analiz, FTIR ve şişme testi ile karakterize edilmiştir. SEM analizleri kriyojellerin, donmuş su kristallerinin porojen etkisi ile oluşan birbirine bağlı 10-50 μm boyutunda gözenekleri ile heterogözenekli bir yapıya sahip olduğunu göstermiştir. Maksimum DNA adsorpsiyon miktarı pH 8’de 19.97 mg/g polymer olarak bulunmuştur

Keywords

• Afinite kromatografi,DNA adsorpsiyon,metal şelat,kriyojeller

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