{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,9,10]],"date-time":"2026-09-10T13:20:40Z","timestamp":1789046440384,"version":"build-2803163510"},"reference-count":91,"publisher":"Wiley","license":[{"start":{"date-parts":[[2026,9,10]],"date-time":"2026-09-10T00:00:00Z","timestamp":1788998400000},"content-version":"vor","delay-in-days":0,"URL":"http:\/\/creativecommons.org\/licenses\/by\/4.0\/"},{"start":{"date-parts":[[2026,9,10]],"date-time":"2026-09-10T00:00:00Z","timestamp":1788998400000},"content-version":"tdm","delay-in-days":0,"URL":"http:\/\/doi.wiley.com\/10.1002\/tdm_license_1.1"}],"content-domain":{"domain":["onlinelibrary.wiley.com"],"crossmark-restriction":true},"short-container-title":["Small"],"abstract":"<jats:title>ABSTRACT<\/jats:title>\n                  <jats:p>\n                    Nanoporous architectures offer unique opportunities to spatially organize biocatalysts while preserving efficient interfacial charge transfer. Here, we introduce a tiered enzyme\u2013electrode interface based on nanoporous gold (np\u2010Au) that enables high\u2010density, activity\u2010retaining immobilization of lactate oxidase for sensitive\n                    <jats:sc>l<\/jats:sc>\n                    \u2010lactate detection. The np\u2010Au scaffold, prepared by controlled dealloying of Ag\u2010Au, exhibits average pore diameter of \u223c170\u00a0nm, providing geometric confinement compatible with the tetrameric structure of\n                    <jats:italic>\u00a0Aerococcus viridans<\/jats:italic>\n                    \u00a0\n                    <jats:sc>l<\/jats:sc>\n                    \u2010lactate oxidase (LOx). A mixed self\u2010assembled monolayer composed of 1\u2010thioglycerol and Ni\n                    <jats:sup>2<\/jats:sup>\n                    <jats:sup>+<\/jats:sup>\n                    \u2010nitrilotriacetic acid\u2010terminated 12\u2010thiododecaneamide establishes a bifunctional interface that combines hydrophilic\u2010antifouling properties with site\u2010directed immobilization of N\u2010terminal His\u2010tagged LOx. This architecture affords high immobilization yield (64%), elevated surface loading (7.44\u00a0mg m\n                    <jats:sup>\u2212<\/jats:sup>\n                    <jats:sup>2<\/jats:sup>\n                    ; \u223c9\u00a0mg g\n                    <jats:sup>\u22121<\/jats:sup>\n                    ), and superior retention of enzymatic activity. Electrochemical impedance spectroscopy reveals controlled modulation of interfacial charge\u2010transfer resistance upon sequential SAM formation and enzyme coupling, confirming the formation of a structurally defined biointerface. The resulting bioelectrode exhibits stable and sensitive amperometric\n                    <jats:sc>l<\/jats:sc>\n                    \u2010lactate detection with a linear dynamic range of 0.12 \u2013 3.0\u00a0mM and a sensitivity of 75\u00a0\u00b5A mM\n                    <jats:sup>\u22121<\/jats:sup>\n                    cm\n                    <jats:sup>\u2212<\/jats:sup>\n                    <jats:sup>2<\/jats:sup>\n                    , covering clinically relevant concentrations in saliva and blood. The presented tiered design strategy establishes a modular platform for high\u2010performance nanoporous metal bioelectrodes with controlled enzyme orientation and enhanced operational stability.\n                  <\/jats:p>","DOI":"10.1002\/smll.75492","type":"journal-article","created":{"date-parts":[[2026,9,10]],"date-time":"2026-09-10T12:06:01Z","timestamp":1789041961000},"update-policy":"https:\/\/doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":0,"title":["Nanopore and Self\u2010Assembled Monolayer Engineering of\n                    <scp>l<\/scp>\n                    \u2010Lactate Oxidase Gold Electrodes for Enhanced\n                    <scp>l<\/scp>\n                    \u2010Lactate Biosensing"],"prefix":"10.1002","author":[{"ORCID":"https:\/\/orcid.org\/0000-0002-4582-165X","authenticated-orcid":false,"given":"Mislav","family":"Su\u0161ac","sequence":"first","affiliation":[{"name":"Institute of Biotechnology and Biochemical Engineering Graz 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Biochemical Engineering Graz University of Technology  Graz Austria"},{"name":"Institute of Materials Physics Graz University of Technology Graz Austria"},{"name":"Infineon Technologies Austria AG \u2013 Development Center Graz  Graz Austria"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-3246-5630","authenticated-orcid":false,"given":"Chao","family":"Zhong","sequence":"additional","affiliation":[{"name":"Institute of Biotechnology and Biochemical Engineering Graz University of Technology  Graz Austria"},{"name":"Austrian Centre of Industrial Biotechnology (acib)  Graz Austria"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-4624-4433","authenticated-orcid":false,"given":"Roland","family":"W\u00fcrschum","sequence":"additional","affiliation":[{"name":"Institute of Materials Physics Graz University of Technology Graz 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