Hacettepe University, Department of Biology, Ankara, Turkey.
Hacettepe University, Department of Chemistry, Ankara, Turkey.
Colloids Surf B Biointerfaces. 2018 Feb 1;162:146-153. doi: 10.1016/j.colsurfb.2017.11.044. Epub 2017 Nov 20.
Boronate affinity chromatography is a common purification method used for isolation and purification of cis-diol containing biomolecules. Poly (hydroxyethyl methacrylate-co-vinyl phenyl boronic acid) [P(HEMA-VPBA)] nanoparticles were prepared by miniemulsion polymerization to use in RNA purification methods. The P(HEMA-VPBA) nanoparticles were characterized by particle size distribution, surface area, Fourier transform infrared spectroscopy and atomic force microscopy. The effects of temperature, pH, RNA concentration and different salt types on RNA binding on the P(HEMA-VPBA) nanoparticles were examined. It was observed that RNA binding was increased with the increasing of pH and max RNA binding was obtained at pH 9.0. RNA binding capacity of the P(HEMA-VPBA) nanoparticles was increased from 167mg/g to 601mg/g with addition of BaCl to the binding medium. Maximum RNA binding capacity of the P(HEMA-VPBA) nanoparticles was 172mg/g at 1.0mg/mL initial RNA concentration. The P(HEMA-VPBA) nanoparticles were reusable for RNA binding. RNA was also extracted from Escherichia coli cells and purified successfully using the P(HEMA-VPBA) nanoparticles.
硼亲和层析是一种常用的纯化方法，用于分离和纯化含有顺二醇的生物分子。通过细乳液聚合制备了聚（羟乙基甲基丙烯酸酯-共-乙烯基苯硼酸）[P（HEMA-VPBA）]纳米粒子，用于 RNA 纯化方法。通过粒径分布、比表面积、傅里叶变换红外光谱和原子力显微镜对 P（HEMA-VPBA）纳米粒子进行了表征。考察了温度、pH 值、RNA 浓度和不同盐类对 P(HEMA-VPBA)纳米粒子上 RNA 结合的影响。结果表明，随着 pH 值的升高，RNA 结合增加，在 pH 9.0 时达到最大结合。在结合介质中加入 BaCl2 后，P(HEMA-VPBA)纳米粒子的 RNA 结合容量从 167mg/g 增加到 601mg/g。在初始 RNA 浓度为 1.0mg/mL 时，P(HEMA-VPBA)纳米粒子的最大 RNA 结合容量为 172mg/g。P(HEMA-VPBA)纳米粒子可重复用于 RNA 结合。还使用 P(HEMA-VPBA)纳米粒子成功地从大肠杆菌细胞中提取和纯化了 RNA。
