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One-step purification and immobilization of his-tagged protein via Ni2+-functionalized Fe3O4@polydopamine magnetic nanoparticles

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Abstract

Ni2+-functionalized Fe3O4@polydopamine magnetic nanoparticles (Ni2+-PD-MNPs) were designed and synthesized by in situ coating of magnetic nanoparticles with polydopamine, followed by conjugation of Ni2+ to the polydopamine film. The Ni2+-PD-MNPs were used to purify His-tagged red fluorescent protein (His-RFP) via affinity interaction between Ni2+ and the His-tag. The results showed that the Ni2+-PD-MNPs had extraordinary selectivity for His-RFP purification. In addition, a Histagged transaminase (ω-transaminase BJ110) was selectively immobilized onto the Ni2+-PD-MNPs without purification, and the immobilized enzyme showed improved specific activity, as well as enhanced stability and reusability.

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Authors and Affiliations

  1. State Key Laboratory of Bioreactor Engineering, New World Institute of Biotechnology, East China University of Science and Technology, Shanghai, 200-237, China

    Jianbing Yang, Kefeng Ni, Dongzhi Wei & Yuhong Ren

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  1. Jianbing Yang
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  2. Kefeng Ni
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  3. Dongzhi Wei
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  4. Yuhong Ren
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Correspondence to Yuhong Ren.

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Yang, J., Ni, K., Wei, D. et al. One-step purification and immobilization of his-tagged protein via Ni2+-functionalized Fe3O4@polydopamine magnetic nanoparticles. Biotechnol Bioproc E 20, 901–907 (2015). https://doi.org/10.1007/s12257-015-0136-7

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  • DOI: https://doi.org/10.1007/s12257-015-0136-7

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