RNA purification from Escherichia coli cells using boronated nanoparticles


PERÇİN DEMİRÇELİK I. , İDİL N. , DENİZLİ A.

COLLOIDS AND SURFACES B-BIOINTERFACES, cilt.162, ss.146-153, 2018 (SCI İndekslerine Giren Dergi) identifier identifier identifier

  • Cilt numarası: 162
  • Basım Tarihi: 2018
  • Doi Numarası: 10.1016/j.colsurfb.2017.11.044
  • Dergi Adı: COLLOIDS AND SURFACES B-BIOINTERFACES
  • Sayfa Sayıları: ss.146-153

Özet

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 167 mg/g to 601 mg/g with addition of BaCl2 to the binding medium. Maximum RNA binding capacity of the P(HEMA-VPBA) nanoparticles was 172 mg/g at 1.0 mg/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. (C) 2017 Elsevier B.V. All rights reserved.