Univerzita Pardubice Fakulta chemicko- technologická

Univerzita Pardubice

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Publikace detail

Unconventional purification of His-tagged proteins using TiO2 Nanotubes Decorated by Fe3O4 Nanoparticles

Autoři: Kupčík Rudolf | Macák Jan | Řehulka Pavel | Sopha Hanna Ingrid | Bílková Zuzana

Rok: 2018

Druh publikace: ostatní - přednáška nebo poster

Strana od-do: nestránkováno

Tituly:

Jazyk	Název	Abstrakt	Klíčová slova
eng	Unconventional purification of His-tagged proteins using TiO2 Nanotubes Decorated by Fe3O4 Nanoparticles	Synthesis of recombinant proteins is important in many fields in the life sciences. The efficient purification is crucial for recombinant biomolecules as they come contaminated with proteins of host cells. Chromatographic step, especially affinity chromatography, is commonly employed for protein purification. One of most utilized techniques, Immobilized-Metal Affinity Chromatography (IMAC), exploits affinity of chelated metal ions with His-tagged recombinant proteins. Unfortunately, however, purity and applicability of such purified protein is limited due to low selectivity of IMAC and toxicity of released ions. The aim of our work is to demonstrate the unique characteristics of TiO2 nanotubes decorated by Fe3O4 nanoparticles with higher selectivity for His-tagged proteins in comparison to commercial IMAC particles. The initial sample, cells lysate containing His-tagged recombinant protein, was used for both materials and separation fractions were analyzed with polyacrylamide gel electrophoresis. The presented TiO2 nanotubes decorated with Fe3O4 nanoparticles showed an enhanced selectivity for His-tagged recombinant proteins demonstrated using gel electrophoresis. Compared to IMAC system, specific isolation of selected His-tagged proteins on behalf of other proteins was significantly improved and presence of contamination was reduced to a minimum. Magnetic properties of material provide simple and quantitative separation of material form analyte. By testing of toxicity and mutagenicity of TiO2 nanotubes, the applicability of this material for applications with high demands on biocompatibility was confirmed. Higher selectivity and magnetic properties allow to obtain highly pure recombinant proteins.	His-tagged proteins; TiO2; Fe3O4; purification