Nickel nanoparticles synthesized by a modified polyol method for the purification of histidine-tagged single-domain antibody ToxA5.1

Nickel nanoparticles (NNPs) synthesized by a modified polyol method using ethylene glycol as a reducing agent, palladium chloride as a nucleating agent, and polyvinylpyrrolidone (PVP) as a protective agent were investigated as a potential magnetic adsorbent for the purification of hexahistidine-tagged (His6-tagged) recombinant proteins. The synthesis resulted in nanoparticles having an average diameter of 68 ± 28 nm. The x-ray diffraction pattern confirmed the presence of nickel metal, as well as the presence of unreacted nickel (II) hydroxide Ni(OH)2. Magnetic characterization showed that a magnetization saturation of 39.3 electromagnetic unit (emu)/g at 20,000 Oersted (Oe) was reached rapidly and that the material exhibited ferromagnetic behavior. Protein purification results showed that the synthesized NNPs were highly selective for binding to a His6-tagged recombinant protein single-domain antibody ToxA5.1. In addition, NNPs were used for four adsorption cycles without significant binding capacity losses. These particles have shown great potential such as being easily synthesized, cost-effective, and highly selective magnetic adsorbents for the purification of His6-tagged recombinant proteins.