Köse Kazım, Erol Kadir, Özgür Erdoğan, Uzun Lokman, Denizli Adil
Hacettepe University, Department of Chemistry, Ankara, Turkey; Hitit University, Scientific Technical Research and Application Center, Çorum, Turkey.
Hacettepe University, Department of Chemistry, Ankara, Turkey; Hitit University, Department of Chemistry, Çorum, Turkey.
Colloids Surf B Biointerfaces. 2016 Oct 1;146:678-86. doi: 10.1016/j.colsurfb.2016.07.003. Epub 2016 Jul 4.
Cryogels are used effectively for many diverse applications in a variety of fields. The isolation or purification of RNA, one of the potential utilizations for cryogels, is crucial due to their vital roles such as encoding, decoding, transcription and translation, and gene expression. RNA principally exists within every living thing, but their tendency to denaturation easily is still the most challenging issue. Herein, we aimed to develop adenine incorporated polymeric cryogels as an alternative sorbent for cost-friendly and fast RNA purification with high capacity. For this goal, we synthesized the polymerizable derivative of adenine called as adenine methacrylate (AdeM) through the substitution reaction between adenine and methacryloyl chloride. Then, 2-hydroxyethyl methacrylate (HEMA)-based cryogels were prepared in a partially frozen aqueous medium by copolymerization of monomers, AdeM, and HEMA. The cryogels were characterized by using Fourier transform infrared spectroscopy (FTIR), scanning electron microscopy (SEM), surface area measurements, thermogravimetric analysis (TGA), and swelling tests. RNA adsorption experiments were performed via batch system while varying different conditions including pH, initial RNA concentration, temperature, and interaction time. We achieved high RNA adsorption capacity of cryogels, with the swelling ratio around 510%, as 11.86mg/g. The cryogels might be reused at least five times without significant decrease in adsorption capacity.
冷冻凝胶在多个领域的众多不同应用中得到了有效利用。RNA的分离或纯化是冷冻凝胶的潜在用途之一，由于RNA在编码、解码、转录、翻译和基因表达等重要过程中发挥着关键作用，因此其分离纯化至关重要。RNA主要存在于每一个生物体内，但其易于变性的特性仍然是最具挑战性的问题。在此，我们旨在开发一种含腺嘌呤的聚合冷冻凝胶，作为一种替代吸附剂，以实现成本友好、快速且具有高容量的RNA纯化。为了实现这一目标，我们通过腺嘌呤与甲基丙烯酰氯之间的取代反应合成了腺嘌呤的可聚合衍生物，即甲基丙烯酸腺嘌呤（AdeM）。然后，在部分冷冻的水介质中，通过单体AdeM和甲基丙烯酸2-羟乙酯（HEMA）的共聚制备了基于HEMA的冷冻凝胶。通过傅里叶变换红外光谱（FTIR）、扫描电子显微镜（SEM）、表面积测量、热重分析（TGA）和溶胀试验对冷冻凝胶进行了表征。在不同条件下，包括pH值、初始RNA浓度、温度和相互作用时间，通过分批系统进行了RNA吸附实验。我们获得了冷冻凝胶对RNA的高吸附容量，溶胀率约为510%，吸附量为11.86mg/g。冷冻凝胶至少可以重复使用五次，而吸附容量不会显著降低。
