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Unlike previous studies that require multi-step surface functionalization with chelating ligands, such as nitrilotriacetic acid (abridged as NTA), to achieve histidine-tagged (denoted as His-tagged) protein binding, this study reports a one-step hydrothermal synthesis of Ni NPs with a spherical-spiky architecture that provides abundant intrinsic Ni2+ active sites. A series of synthesis experiments were conducted to systematically optimize the hydrothermal reaction conditions (solution pH, temperature, and time). The ability of Ni NPs to separate and purify His-tagged fusion proteins was evaluated in relation to their abundant Ni2+ active binding sites and robust magnetic responsiveness. It was found that Ni NPs were able to rapidly, specifically, and efficiently purify a variety of His-tagged proteins, achieving a high saturation adsorption capacity of 80.36 mg\/g, significantly higher than commercial Ni-NTA resins and previously reported Ni-based adsorbents, while retaining excellent recyclability (&amp;gt;80% after 5 cycles). These features underscore the potential of Ni NPs as cost-effective, reusable, and high-performance nano-adsorbents for protein separation, offering distinct advantages over conventional functionalization-dependent methods.<\/jats:p>","DOI":"10.1021\/acsabm.6c00399","type":"journal-article","created":{"date-parts":[[2026,7,27]],"date-time":"2026-07-27T08:01:31Z","timestamp":1785139291000},"page":"7409-7418","source":"Crossref","is-referenced-by-count":0,"title":["Preparation of Magnetic Nickel Nanoparticles and Their Use in Efficient Purification of Histidine-Tagged Fusion Proteins"],"prefix":"10.1021","volume":"9","author":[{"given":"Mochou","family":"Gao","sequence":"first","affiliation":[{"name":"Henan University , , ,","place":["Zhengzhou, China, 450046"]},{"name":"Jilin University , , ,","place":["Changchun, China, 130012"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Xing","family":"Miao","sequence":"additional","affiliation":[{"name":"Henan University , , ,","place":["Zhengzhou, China, 450046"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Haowen","family":"Ren","sequence":"additional","affiliation":[{"name":"Xinjiang Agricultural University , , ,","place":["Urumqi, China, 830052"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Haocheng","family":"Xu","sequence":"additional","affiliation":[{"name":"Xi'an Jiaotong-Liverpool University , , ,","place":["Suzhou, China, 215000"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Haotian","family":"Li","sequence":"additional","affiliation":[{"name":"Henan University , , ,","place":["Zhengzhou, China, 450046"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Hao","family":"Wang","sequence":"additional","affiliation":[{"name":"Henan University , , ,","place":["Zhengzhou, China, 450046"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Kehao","family":"Zhu","sequence":"additional","affiliation":[{"name":"Henan University , , ,","place":["Zhengzhou, China, 450046"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Chenglin","family":"Xu","sequence":"additional","affiliation":[{"name":"Henan University , , ,","place":["Zhengzhou, China, 450046"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Yanbin","family":"Yin","sequence":"additional","affiliation":[{"name":"Henan University , , ,","place":["Zhengzhou, China, 450046"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Yanbao","family":"Zhao","sequence":"additional","affiliation":[{"name":"Henan University , , ,","place":["Zhengzhou, China, 450046"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Wensheng","family":"Yang","sequence":"additional","affiliation":[{"name":"Henan University , , ,","place":["Zhengzhou, China, 450046"]},{"name":"Jilin University , , ,","place":["Changchun, China, 130012"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-0373-7203","authenticated-orcid":true,"given":"Xueyan","family":"Zou","sequence":"additional","affiliation":[{"name":"Henan University , , ,","place":["Zhengzhou, China, 450046"]}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"316","published-online":{"date-parts":[[2026,7,25]]},"reference":[{"issue":"5","key":"2026081707153702000_cit1","doi-asserted-by":"publisher","first-page":"267","DOI":"10.1038\/s41580-018-0092-0","article-title":"Mitochondrial proteins: from biogenesis to functional networks","volume":"20","author":"Pfanner","year":"2019","journal-title":"Nat. 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