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Starch-coated magnetic iron oxide nanoparticles for affinity purification of recombinant proteins / V. V. Krasitskaya, A. N. Kudryavtsev, R. N. Yaroslavtsev [et al.] // Int. J. Mol. Sci. - 2022. - Vol. 23, Is. 10. - Ст. 5410, DOI 10.3390/ijms23105410. - Cited References: 37. - This study was supported by the Russian Science Foundation and the Krasnoyarsk Territorial Foundation for Support of Scientific and R&D Acitvities, project No. 22-14-20020
. - ISSN 1661-6596
Кл.слова (ненормированные):
iron oxide nanoparticles -- starch -- affinity sorbent -- maltose-binding protein -- hybrid proteins purification
Аннотация: Starch-coated magnetic iron oxide nanoparticles have been synthesized by a simple, fast, and cost-effective co-precipitation method with cornstarch as a stabilizing agent. The structural and magnetic characteristics of the synthesized material have been studied by transmission electron microscopy, Mossbauer spectroscopy, and vibrating sample magnetometry. The nature of bonds between ferrihydrite nanoparticles and a starch shell has been examined by Fourier transform infrared spectroscopy. The data on the magnetic response of the prepared composite particles have been obtained by magnetic measurements. The determined magnetic characteristics make the synthesized material a good candidate for use in magnetic separation. Starch-coated magnetic iron oxide nanoparticles have been tested as an affinity sorbent for one-step purification of several recombinant proteins (cardiac troponin I, survivin, and melanoma inhibitory activity protein) bearing the maltose-binding protein as an auxiliary fragment. It has been shown that, due to the highly specific binding of this fragment to the starch shell, the target fusion protein is selectively immobilized on magnetic nanoparticles and eluted with the maltose solution. The excellent efficiency of column-free purification, high binding capacity of the sorbent (100–500 µg of a recombinant protein per milligram of starch-coated magnetic iron oxide nanoparticles), and reusability of the obtained material have been demonstrated.
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Держатели документа:
Institute of Biophysics SB RAS, Federal Research Center “Krasnoyarsk Science Center SB RAS”, Krasnoyarsk, 660036, Russian Federation
Kirensky Institute of Physics SB RAS, Federal Research Center “Krasnoyarsk Science Center SB RAS”, Krasnoyarsk, 660036, Russian Federation
Federal Research Center “Krasnoyarsk Science Center SB RAS”, Krasnoyarsk, 660036, Russian Federation
School of Fundamental Biology and Biotechnology, Siberian Federal University, Krasnoyarsk, 660041, Russian Federation
Доп.точки доступа:
Krasitskaya, V. V.; Kudryavtsev, A. N.; Yaroslavtsev, R. N.; Ярославцев, Роман Николаевич; Gerasimova, Yu. V.; Герасимова, Юлия Валентиновна; Velikanov, D. A.; Великанов, Дмитрий Анатольевич; Bayukov, O. A.; Баюков, Олег Артемьевич; Stolyar, S. V.; Столяр, Сергей Викторович; Frank, L. A.
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