TY - JOUR
T1 - Chitosan-coated magnetic iron oxide nanoparticles for DNA and rhEGF separation
AU - Gómez Pérez, Annia
AU - González-Martínez, Eduardo
AU - Díaz Águila, Carlos R.
AU - González-Martínez, David A.
AU - González Ruiz, Gustavo
AU - García Artalejo, Aymed
AU - Yee-Madeira, Hernani
N1 - Publisher Copyright:
© 2020 Elsevier B.V.
PY - 2020/4/20
Y1 - 2020/4/20
N2 - The obtention and purification of DNA and recombinant proteins are critical steps in the biotech industries. In this research, the use of chitosan-coated magnetic iron oxide nanoparticles as magnetic nano-adsorbent was investigated. Iron oxide nanoparticles were obtained through a simple coprecipitation method. The spinel structure of the nanoparticles was confirmed by X-Ray diffraction analysis. The particle size before (16 nm) and after chitosan coating (14 nm) was measured using scanning electron microscopy. Infrared spectroscopy and thermogravimetric analysis measurements confirmed the presence of chitosan on the surface of magnetic nanoparticles coated in a percentage of 11.24%. The Redlich-Peterson isotherm yielded the best fit for the DNA experimental adsorption capacity and a maximum of 98 mg/g was obtained. The structural integrity of DNA, after the elution process, was confirmed by agarose gel electrophoresis. An adsorption capacity of 440 mg/g for rhEGF was found and the Langmuir-Freundlich isotherm showed the best fit for the experimental results. Finally, SDS-PAGE and Western blot assays confirmed that the adsorption/desorption process did not affect the rhEGF identity, thereby, suggesting that the biological activity was preserved.
AB - The obtention and purification of DNA and recombinant proteins are critical steps in the biotech industries. In this research, the use of chitosan-coated magnetic iron oxide nanoparticles as magnetic nano-adsorbent was investigated. Iron oxide nanoparticles were obtained through a simple coprecipitation method. The spinel structure of the nanoparticles was confirmed by X-Ray diffraction analysis. The particle size before (16 nm) and after chitosan coating (14 nm) was measured using scanning electron microscopy. Infrared spectroscopy and thermogravimetric analysis measurements confirmed the presence of chitosan on the surface of magnetic nanoparticles coated in a percentage of 11.24%. The Redlich-Peterson isotherm yielded the best fit for the DNA experimental adsorption capacity and a maximum of 98 mg/g was obtained. The structural integrity of DNA, after the elution process, was confirmed by agarose gel electrophoresis. An adsorption capacity of 440 mg/g for rhEGF was found and the Langmuir-Freundlich isotherm showed the best fit for the experimental results. Finally, SDS-PAGE and Western blot assays confirmed that the adsorption/desorption process did not affect the rhEGF identity, thereby, suggesting that the biological activity was preserved.
KW - DNA recovery
KW - Protein recovery
KW - chitosan-coated magnetic iron oxide nanoparticles
UR - http://www.scopus.com/inward/record.url?scp=85080072556&partnerID=8YFLogxK
U2 - 10.1016/j.colsurfa.2020.124500
DO - 10.1016/j.colsurfa.2020.124500
M3 - Artículo
AN - SCOPUS:85080072556
SN - 0927-7757
VL - 591
JO - Colloids and Surfaces A: Physicochemical and Engineering Aspects
JF - Colloids and Surfaces A: Physicochemical and Engineering Aspects
M1 - 124500
ER -