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Here, we report the development, systematic optimization and automated implementation of an amine-functionalized NiFe\n                    <jats:sub>2<\/jats:sub>\n                    O\n                    <jats:sub>4<\/jats:sub>\n                    -based MNP protocol for\n                    <jats:italic>N<\/jats:italic>\n                    -glycan purification. Critical workflow steps, including particle drying and dispersion, binding efficiency, and elution buffer selection, were comprehensively evaluated. The optimized protocol, employing 0.5\u00a0mg\/mL NiFe\n                    <jats:sub>2<\/jats:sub>\n                    O\n                    <jats:sub>4<\/jats:sub>\n                    -NH\n                    <jats:sub>2<\/jats:sub>\n                    particles and 150 mM ammonium-formate (pH 4.4) as elution buffer enabled the reproducible identification of the major\n                    <jats:italic>N<\/jats:italic>\n                    -glycan peaks from human serum and immunoglobulin G (IgG). Importantly, the method was successfully translated to a Hamilton Microlab Prep robotic liquid handling platform in a 96-well format, ensuring scalability and reduced manual variability. Overall, this study advances a previously established MNP-based glycan purification approach into a robust, scalable, and automation-compatible workflow through synthesis redesign, systematic optimization, and successful robotic implementation.\n                  <\/jats:p>","DOI":"10.1038\/s41598-026-54879-1","type":"journal-article","created":{"date-parts":[[2026,6,8]],"date-time":"2026-06-08T14:38:12Z","timestamp":1780929492000},"update-policy":"https:\/\/doi.org\/10.1007\/springer_crossmark_policy","source":"Crossref","is-referenced-by-count":0,"title":["Automation of N-glycan purification using amine-functionalized NiFe2O4 magnetic nanoparticles"],"prefix":"10.1038","volume":"16","author":[{"given":"Dalma","family":"Dojcs\u00e1k","sequence":"first","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"\u00c1gnes M\u00e1ria","family":"Ilosvai","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"L\u00e1szl\u00f3","family":"Vanyorek","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Csaba","family":"V\u00e1radi","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"297","published-online":{"date-parts":[[2026,6,8]]},"reference":[{"key":"54879_CR1","doi-asserted-by":"publisher","first-page":"529","DOI":"10.1146\/annurev-biochem-030122-044347","volume":"93","author":"N de Haan","year":"2024","unstructured":"de Haan, N., Nielsen, M. 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The study protocol was approved by the Regional Research Ethics Committee (approval number: RKEB\/IKEB-G-102-102-2018).","order":2,"name":"Ethics","label":"Ethical approval","group":{"name":"EthicsHeading","label":"Declarations"}},{"value":"This study was approved by the Regional Research Ethics Committee under approval number RKEB\/IKEB-G-102-102-2018.","order":3,"name":"Ethics","label":"Institutional review board statement","group":{"name":"EthicsHeading","label":"Declarations"}},{"value":"Written informed consent forms were obtained from all the patients in accordance with the Declaration of Helsinki.","order":4,"name":"Ethics","label":"Informed consent statement","group":{"name":"EthicsHeading","label":"Declarations"}},{"value":"Samples of the nanoparticles are available from the authors.","order":5,"name":"Ethics","label":"Sample availability","group":{"name":"EthicsHeading","label":"Declarations"}}],"article-number":"26094"}}