Synthesis of Ni2+-functionalized polydopamine magnetic beads for facilitated purification of histidine-tagged proteins

AMB Express

Original Article

Alireza Shariati¹  Sara Ali Hosseinzadeh²  Kosar Atarod³  Sogand Sadat Mortazavi³  Zahra Barghi⁴  Fatemeh Sadat Shariati⁴  Behrokh Farahmand⁴ 

[1] Department of Materials Engineering, Tarbiat Modares University, P.O. Box 14115-143, Tehran, Iran;Department of Nanobiotechnology, New Technologies Research Group, Pasteur Institute of Iran, Tehran, Iran;Farzanegan high school, National Organization for Development of Exceptional Talents, Tehran, Iran;Infuenza Research Lab, Pasteur Institute of Iran, Tehran, Iran;

关键词: Magnetic beads;    Facillated purification;    Polydopamine;    His-tagged protein;    Recombinant protein;   

DOI  :  10.1186/s13568-023-01613-z

 received in 2023-07-01, accepted in 2023-09-24,  发布年份 2023

来源: Springer

【 摘 要 】

Facilitated purification of proteins, at a low cost and a short time, is one of the key steps in the industrial production of recombinant proteins. In the current study, polydopamine nanoparticles (PDA-NPs) are considered in the synthesis of magnetic beads for purifying recombinant proteins due to advantages such as biocompatibility/ biodegradability, easy synthesis, as well as the ability to directly chelate metal ions. They were synthesized in Tris buffer (pH: 8:5), then chelated with Fe3+(20 mg) and Ni2+ ions at concentrations of 2, 3, 5, and 7 mg/ml. Prepared nanoparticles were characterized through scanning electron microscopy (SEM), ultraviolet-visible spectroscopy (UV-vis), dynamic light scattering (DLS), Inductively Coupled Plasma (ICP), and vibrating sample magnetometer (VSM). The size distribution of the particles was reported in the narrow range of 120–140 nm and 200 to 220 nm by the SEM image and DLS analysis, respectively. The chelation of ions on the surface of the nanoparticle was confirmed by the ICP technique with a magnetization of 35.42 emu/g. The highest adsorption rate of Ni2+ ions to polydopamine was obtained at a ratio of 1.4. The SDS-PAGE and western blot analysis confirmed the purification of eGFP and Hsp40 by PDA/Fe3+/Ni2+ at 26 and 40 kDa compared to the commercial nickel column. Moreover, the concentration of purified eGFP by PDA/Fe3+/Ni2+ was reported 138.83 µg/ml by the fluorescent signals, which is almost equal to or more than the protein purified by commercial Ni-NTA column (108.28 µg/ ml). The stability of PDA/Fe3+/Ni2+ has also been evaluated by ICP-OES for 10 days, and the result suggested that PDA magnetic beads were stable. Therefore, it can be concluded that PDA/Fe3+/Ni2+ have the ability to purify recombinant proteins in one less step and shorter time.

【 授权许可】

CC BY   
© Springer-Verlag GmbH Germany, part of Springer Nature 2023

【 图 表 】

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