Abstract
Nano-magnetite with superparamagnetism could be coated by some organic compounds or by nano Au or Pt via surface modifications with multi-step reactions for the applications of isolating histidine-tagged (His-tagged) proteins. Introducing active sites of binding histidine onto the surface of nano-magnetite was the ultimate task. However, multi-step treatments might result in departure of the coatings from the surface of the nano-magnetite, which led to loss of active sites. In this work, we reported a convenient and efficient way of treating nano-magnetites and applied them in isolating His-tagged proteins. Carboxylates were introduced on the surface of home-made nano-magnetite directly via ultrasonic mixing with sodium bitartrate rather than complicated surface modifications, which was proved by thermogravimetric analyses. Ni2+ was, therefore, caught by the carboxylates of the coating via the coordinate interaction, demonstrated by X-ray photoelectron spectra. The coated magnetic nanoparticles with the bonded Ni2+ were successfully employed to selectively bind and separate recombinant His-tagged proteins directly from the mixture of Escherichia coli cell lysate, and showed wonderful affinity for His-tagged proteins with the saturated adsorption amount being 556mgg-1. Additionally, such functionalized nano-magnetite manifested the excellent recyclability in isolating His-tagged proteins.
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