Light-Driven Biosensor for the Rapid and Selective Detection of Hypoxia-Inducible Factor – Prolyl Hydroxylase Domain Inhibitors, in Aqueous Media and Saliva

Light-Driven Biosensor for the Rapid and Selective Detection of Hypoxia-Inducible Factor – Prolyl Hydroxylase Domain Inhibitors, in Aqueous Media and Saliva | Research Square window.SnipcartSettings = { analytics: { enabled: false } }; (function() { var accessVector = localStorage.getItem('access_vector') || ''; window.dataLayer = window.dataLayer || []; if (accessVector) { window.dataLayer.push({ user: { profile: { profileInfo: { snid: accessVector } } } }); } })(); (function(w,d,s,l,i){w[l]=w[l]||[];w[l].push({'gtm.start':new Date().getTime(),event:'gtm.js'});var f=d.getElementsByTagName(s)[0],j=d.createElement(s),dl=l!='dataLayer'?'&l='+l:'';j.async=true;j.src='https://www.googletagmanager.com/gtm.js?id='+i+dl;f.parentNode.insertBefore(j,f);})(window,document,'script','dataLayer','GTM-K279D39R'); Browse Preprints In Review Journals COVID-19 Preprints AJE Video Bytes Research Tools Research Promotion AJE Professional Editing AJE Rubriq About Preprint Platform In Review Editorial Policies Our Team Advisory Board Help Center Sign In Submit a Preprint Cite Share Download PDF Research Article Light-Driven Biosensor for the Rapid and Selective Detection of Hypoxia-Inducible Factor – Prolyl Hydroxylase Domain Inhibitors, in Aqueous Media and Saliva Rebecca. L. Houston, Godwin Ayoko, Emad. L. Izake This is a preprint; it has not been peer reviewed by a journal. https://doi.org/ 10.21203/rs.3.rs-7172116/v1 This work is licensed under a CC BY 4.0 License Status: Published Journal Publication published 02 Oct, 2025 Read the published version in Microchimica Acta → Version 1 posted 9 You are reading this latest preprint version Abstract Hypoxia-inducible factor prolyl hydroxylase domain inhibitors (HIF-PHI) are to increase the production of EPO and haemoglobin in patients with chronic kidney disease. HIF inhibitors such as Molidustat and Vadadustat are used as doping agents in racing sports. In this work we present new sensitive and selective materials for the biosensing of Molidustat and Vadadustat in aqueous solution and saliva. Plasmonic nickel foam was synthesised using a reproducible chemical method that deposits forests of closely packed cauliflower-shaped gold nanostructures on the nickel surface. The plasmonic material showed a high enhancement factor of Raman signals (1.09 × 10⁷). Therefore, it was used as a sensor for the detection of HIF inhibitors by surface enhanced Raman spectroscopy (SERS). A target-specific extractor chip for HIF-PHI was fabricated by functionalising the plasmonic nickel foam with Prolyl Hydroxylase Domain-containing protein 2 (PHD2) molecules. The surface functionalisation was carried out by a green synthesis process that utilise deep UV LED light to convert the PHD2 molecule into a bio-thiol that attaches to the plasmonic nickel foam via Au-S bonds. Using the new SERS sensor and extractor chip, Molidustat and Vadadustat were quantified in aqueous solution down to 0.01 µg/L. The new materials were also utilised for the rapid screening of HIF-PHI in spiked saliva samples. The new materials and light-driven surface functionalisation process can be easily adopted to develop new screening methods for multiple biomolecules (e.g. drugs, proteins, disease biomarkers, environmental toxins) for many applications (e.g. therapeutic drug monitoring, doping control, clinical diagnostics environmental monitoring). PHD2 Biosensor Light-driven functionalisation HIF inhibitors doping control Full Text Additional Declarations No competing interests reported. Supplementary Files Supplementarydustats.docx graphicalabstract.tif Cite Share Download PDF Status: Published Journal Publication published 02 Oct, 2025 Read the published version in Microchimica Acta → Version 1 posted Editorial decision: Revision requested 14 Aug, 2025 Reviews received at journal 14 Aug, 2025 Reviews received at journal 07 Aug, 2025 Reviewers agreed at journal 24 Jul, 2025 Reviewers agreed at journal 23 Jul, 2025 Reviewers invited by journal 23 Jul, 2025 Editor assigned by journal 22 Jul, 2025 Submission checks completed at journal 22 Jul, 2025 First submitted to journal 20 Jul, 2025 You are reading this latest preprint version Research Square lets you share your work early, gain feedback from the community, and start making changes to your manuscript prior to peer review in a journal. As a division of Research Square Company, we’re committed to making research communication faster, fairer, and more useful. We do this by developing innovative software and high quality services for the global research community. Our growing team is made up of researchers and industry professionals working together to solve the most critical problems facing scientific publishing. Also discoverable on Platform About Our Team In Review Editorial Policies Advisory Board Help Center Resources Author Services Accessibility API Access RSS feed Manage Cookie Preferences © Research Square 2026 | ISSN 2693-5015 (online) Privacy Policy Terms of Service Do Not Sell My Personal Information {"props":{"pageProps":{"initialData":{"identity":"rs-7172116","acceptedTermsAndConditions":true,"allowDirectSubmit":false,"archivedVersions":[],"articleType":"Research Article","associatedPublications":[],"authors":[{"id":490197042,"identity":"e8669032-19a6-4d4c-9370-b5998d647b46","order_by":0,"name":"Rebecca. L. Houston","email":"data:image/png;base64,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","orcid":"","institution":"Queensland University of Technology (QUT)","correspondingAuthor":true,"prefix":"","firstName":"Rebecca.","middleName":"L.","lastName":"Houston","suffix":""},{"id":490197043,"identity":"e5aa0f43-6edb-4cb1-8787-b5f2e79ab6f7","order_by":1,"name":"Godwin Ayoko","email":"","orcid":"","institution":"Queensland University of Technology (QUT)","correspondingAuthor":false,"prefix":"","firstName":"Godwin","middleName":"","lastName":"Ayoko","suffix":""},{"id":490197044,"identity":"eb167015-d51f-44be-9738-835e80fa4c15","order_by":2,"name":"Emad. L. Izake","email":"","orcid":"","institution":"Queensland University of Technology (QUT)","correspondingAuthor":false,"prefix":"","firstName":"Emad.","middleName":"L.","lastName":"Izake","suffix":""}],"badges":[],"createdAt":"2025-07-21 00:53:18","currentVersionCode":1,"declarations":"","doi":"10.21203/rs.3.rs-7172116/v1","doiUrl":"https://doi.org/10.21203/rs.3.rs-7172116/v1","draftVersion":[],"editorialEvents":[{"content":"https://doi.org/10.1007/s00604-025-07579-y","type":"published","date":"2025-10-02T15:57:40+00:00"}],"editorialNote":"","failedWorkflow":false,"files":[{"id":92883764,"identity":"5ba7a4a8-2176-4bf9-b103-a7fb1dddc7d5","added_by":"auto","created_at":"2025-10-06 16:09:10","extension":"pdf","order_by":1,"title":"","display":"","copyAsset":false,"role":"manuscript-pdf","size":866900,"visible":true,"origin":"","legend":"","description":"","filename":"Manuscriptdustat.pdf","url":"https://assets-eu.researchsquare.com/files/rs-7172116/v1_covered_35147d30-f6a0-4d85-a0ee-b7956d26f6ba.pdf"},{"id":87598114,"identity":"46fa32cf-d6eb-49fa-8f29-8b3129a46eb2","added_by":"auto","created_at":"2025-07-25 16:20:29","extension":"docx","order_by":1,"title":"","display":"","copyAsset":false,"role":"supplement","size":1101310,"visible":true,"origin":"","legend":"","description":"","filename":"Supplementarydustats.docx","url":"https://assets-eu.researchsquare.com/files/rs-7172116/v1/555e54efe0d3b588f624a4cf.docx"},{"id":87598112,"identity":"3feb5583-7bfc-4594-8b28-883b60414ef5","added_by":"auto","created_at":"2025-07-25 16:20:29","extension":"tif","order_by":2,"title":"","display":"","copyAsset":false,"role":"supplement","size":392566,"visible":true,"origin":"","legend":"","description":"","filename":"graphicalabstract.tif","url":"https://assets-eu.researchsquare.com/files/rs-7172116/v1/e6acbad0114edf1c91fc337a.tif"}],"financialInterests":"No competing interests reported.","formattedTitle":"Light-Driven Biosensor for the Rapid and Selective Detection of Hypoxia-Inducible Factor – Prolyl Hydroxylase Domain Inhibitors, in Aqueous Media and Saliva","fulltext":[],"fulltextSource":"","fullText":"","funders":[],"hasAdminPriorityOnWorkflow":false,"hasManuscriptDocX":false,"hasOptedInToPreprint":true,"hasPassedJournalQc":"","hasAnyPriority":false,"hideJournal":false,"highlight":"","institution":"","isAcceptedByJournal":true,"isAuthorSuppliedPdf":true,"isDeskRejected":"","isHiddenFromSearch":false,"isInQc":false,"isInWorkflow":false,"isPdf":true,"isPdfUpToDate":true,"isWithdrawnOrRetracted":false,"journal":{"display":true,"email":"[email protected]","identity":"microchimica-acta","isNatureJournal":false,"hasQc":true,"allowDirectSubmit":false,"externalIdentity":"miac","sideBox":"Learn more about [Microchimica Acta](https://link.springer.com/journal/604)","snPcode":"604","submissionUrl":"https://submission.springernature.com/new-submission/604/3","title":"Microchimica Acta","twitterHandle":"","acdcEnabled":true,"dfaEnabled":true,"editorialSystem":"stoa","reportingPortfolio":"Springer Hybrid","inReviewEnabled":true,"inReviewRevisionsEnabled":false},"keywords":"PHD2 Biosensor, Light-driven functionalisation, HIF inhibitors, doping control","lastPublishedDoi":"10.21203/rs.3.rs-7172116/v1","lastPublishedDoiUrl":"https://doi.org/10.21203/rs.3.rs-7172116/v1","license":{"name":"CC BY 4.0","url":"https://creativecommons.org/licenses/by/4.0/"},"manuscriptAbstract":"\u003cp\u003eHypoxia-inducible factor prolyl hydroxylase domain inhibitors (HIF-PHI) are to increase the production of EPO and haemoglobin in patients with chronic kidney disease. HIF inhibitors such as Molidustat and Vadadustat are used as doping agents in racing sports. In this work we present new sensitive and selective materials for the biosensing of Molidustat and Vadadustat in aqueous solution and saliva. Plasmonic nickel foam was synthesised using a reproducible chemical method that deposits forests of closely packed cauliflower-shaped gold nanostructures on the nickel surface. The plasmonic material showed a high enhancement factor of Raman signals (1.09 \u0026times; 10⁷). Therefore, it was used as a sensor for the detection of HIF inhibitors by surface enhanced Raman spectroscopy (SERS). A target-specific extractor chip for HIF-PHI was fabricated by functionalising the plasmonic nickel foam with Prolyl Hydroxylase Domain-containing protein 2 (PHD2) molecules. The surface functionalisation was carried out by a green synthesis process that utilise deep UV LED light to convert the PHD2 molecule into a bio-thiol that attaches to the plasmonic nickel foam via Au-S bonds. Using the new SERS sensor and extractor chip, Molidustat and Vadadustat were quantified in aqueous solution down to 0.01 \u0026micro;g/L. The new materials were also utilised for the rapid screening of HIF-PHI in spiked saliva samples. The new materials and light-driven surface functionalisation process can be easily adopted to develop new screening methods for multiple biomolecules (e.g. drugs, proteins, disease biomarkers, environmental toxins) for many applications (e.g. therapeutic drug monitoring, doping control, clinical diagnostics environmental monitoring).\u003c/p\u003e","manuscriptTitle":"Light-Driven Biosensor for the Rapid and Selective Detection of Hypoxia-Inducible Factor – Prolyl Hydroxylase Domain Inhibitors, in Aqueous Media and Saliva","msid":"","msnumber":"","nonDraftVersions":[{"code":1,"date":"2025-07-25 16:20:24","doi":"10.21203/rs.3.rs-7172116/v1","editorialEvents":[{"type":"communityComments","content":0},{"type":"decision","content":"Revision requested","date":"2025-08-14T20:14:08+00:00","index":"","fulltext":""},{"type":"editorInvitedReview","content":"","date":"2025-08-14T15:07:04+00:00","index":"hide","fulltext":""},{"type":"editorInvitedReview","content":"","date":"2025-08-07T09:22:40+00:00","index":"hide","fulltext":""},{"type":"reviewerAgreed","content":"234836772303431262497878697675103674492","date":"2025-07-24T08:44:18+00:00","index":"hide","fulltext":""},{"type":"reviewerAgreed","content":"123435497711463314177398056825626865989","date":"2025-07-24T01:58:23+00:00","index":"hide","fulltext":""},{"type":"reviewersInvited","content":"","date":"2025-07-23T18:42:04+00:00","index":"","fulltext":""},{"type":"editorAssigned","content":"","date":"2025-07-22T23:18:50+00:00","index":"","fulltext":""},{"type":"checksComplete","content":"","date":"2025-07-22T23:18:21+00:00","index":"","fulltext":""},{"type":"submitted","content":"Microchimica Acta","date":"2025-07-21T00:50:31+00:00","index":"","fulltext":""}],"status":"published","journal":{"display":true,"email":"[email protected]","identity":"microchimica-acta","isNatureJournal":false,"hasQc":true,"allowDirectSubmit":false,"externalIdentity":"miac","sideBox":"Learn more about [Microchimica Acta](https://link.springer.com/journal/604)","snPcode":"604","submissionUrl":"https://submission.springernature.com/new-submission/604/3","title":"Microchimica Acta","twitterHandle":"","acdcEnabled":true,"dfaEnabled":true,"editorialSystem":"stoa","reportingPortfolio":"Springer Hybrid","inReviewEnabled":true,"inReviewRevisionsEnabled":false}}],"origin":"","ownerIdentity":"581896ab-4325-417d-9269-d2c5d5efb413","owner":[],"postedDate":"July 25th, 2025","published":true,"recentEditorialEvents":[],"rejectedJournal":[],"revision":"","amendment":"","status":"published-in-journal","subjectAreas":[],"tags":[],"updatedAt":"2025-10-06T16:02:34+00:00","versionOfRecord":{"articleIdentity":"rs-7172116","link":"https://doi.org/10.1007/s00604-025-07579-y","journal":{"identity":"microchimica-acta","isVorOnly":false,"title":"Microchimica Acta"},"publishedOn":"2025-10-02 15:57:40","publishedOnDateReadable":"October 2nd, 2025"},"versionCreatedAt":"2025-07-25 16:20:24","video":"","vorDoi":"10.1007/s00604-025-07579-y","vorDoiUrl":"https://doi.org/10.1007/s00604-025-07579-y","workflowStages":[]},"version":"v1","identity":"rs-7172116","journalConfig":"researchsquare"},"__N_SSP":true},"page":"/article/[identity]/[[...version]]","query":{"redirect":"/article/rs-7172116","identity":"rs-7172116","version":["v1"]},"buildId":"8U1c8b4HqxoKbykW_rLl7","isFallback":false,"isExperimentalCompile":false,"dynamicIds":[84888],"gssp":true,"scriptLoader":[]}