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Despite its homologies, the key difference is that AINX is central nervous system-specific. The purpose of this study was to develop an immunoassay for cerebrospinal fluid (CSF) AINX, quantify it in patients with neurodegenerative diseases, and compare its diagnostic performance with neurofilament light (NfL).<\/jats:p>\n                  <\/jats:sec>\n                  <jats:sec>\n                    <jats:title>Methods<\/jats:title>\n                    <jats:p>Monoclonal antibodies were generated and characterized by immunoprecipitation followed by mass spectrometry and ELISA, for specificity to the target analyte and cross-reactivity. Single molecule array (Simoa) technology was the selected platform for assay development. The assay was validated according to pre-established parameters and AINX performance as a biomarker was tested in two independent cohorts (Gothenburg and University College London).<\/jats:p>\n                  <\/jats:sec>\n                  <jats:sec>\n                    <jats:title>Results<\/jats:title>\n                    <jats:p>Two highly specific antibodies were generated (B15 and Ina1) and used to develop a robust assay in all analytical validation parameters tested. The quantitative range of the assay was 0.137\u2013140\u00a0pg\/mL. In the Gothenburg cohort, AINX and NfL showed similar diagnostic performance, but the correlation between biomarkers was diagnosis dependent. In the University College London cohort, AINX and NfL showed similar trends across the different neurodegenerative diseases, with strong correlation between markers.<\/jats:p>\n                  <\/jats:sec>\n                  <jats:sec>\n                    <jats:title>Conclusions<\/jats:title>\n                    <jats:p>We developed a highly sensitive and specific immunoassay for AINX in CSF. AINX showed similar diagnostic performance and high correlation with CSF NfL. Further research should focus on describing its role in specific disorders, as well as evaluate its potential as a blood-based biomarker.<\/jats:p>\n                  <\/jats:sec>","DOI":"10.1007\/s00415-025-13428-y","type":"journal-article","created":{"date-parts":[[2025,10,6]],"date-time":"2025-10-06T11:11:49Z","timestamp":1759749109000},"update-policy":"https:\/\/doi.org\/10.1007\/springer_crossmark_policy","source":"Crossref","is-referenced-by-count":4,"title":["Alpha-internexin as a brain-specific neurodegeneration marker: development and validation of a novel CSF assay"],"prefix":"10.1007","volume":"272","author":[{"ORCID":"https:\/\/orcid.org\/0000-0002-3343-7351","authenticated-orcid":false,"given":"Francisco J.","family":"Meda","sequence":"first","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Anna","family":"Dittrich","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Ingmar","family":"Skoog","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Silke","family":"Kern","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Claire","family":"Leckey","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Edward J.","family":"Wild","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Ross W.","family":"Paterson","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Gunnar","family":"Brinkmalm","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Ulf","family":"Andreasson","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Kaj","family":"Blennow","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Henrik","family":"Zetterberg","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Hlin","family":"Kvartsberg","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"297","published-online":{"date-parts":[[2025,10,6]]},"reference":[{"key":"13428_CR1","doi-asserted-by":"publisher","first-page":"1316","DOI":"10.1083\/jcb.101.4.1316","volume":"101","author":"JS Pachter","year":"1985","unstructured":"Pachter JS, Liem RK (1985) alpha-Internexin, a 66-kD intermediate filament-binding protein from mammalian central nervous tissues. 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All honoraria for these consultancies were paid through the offices of UCL Consultants Ltd., a wholly owned subsidiary of University College London.KB has served as a consultant and at advisory boards for Abbvie, AC Immune, ALZPath, AriBio, Beckman-Coulter, BioArctic, Biogen, Eisai, Lilly, Moleac Pte. Ltd, Neurimmune, Novartis, Ono Pharma, Prothena, Quanterix, Roche Diagnostics, Sunbird Bio, Sanofi, and Siemens Healthineers; has served at data monitoring committees for Julius Clinical and Novartis; has given lectures, produced educational materials and participated in educational programs for AC Immune, Biogen, Celdara Medical, Eisai, and Roche Diagnostics; and is a co-founder of Brain Biomarker Solutions in Gothenburg AB (BBS), which is a part of the GU Ventures Incubator Program, outside the work presented in this paper.HZ has served at scientific advisory boards and\/or as a consultant for Abbvie, Acumen, Alector, Alzinova, ALZpath, Amylyx, Annexon, Apellis, Artery Therapeutics, AZTherapies, Cognito Therapeutics, CogRx, Denali, Eisai, Enigma, LabCorp, Merck Sharp & Dohme, Merry Life, Nervgen, Novo Nordisk, Optoceutics, Passage Bio, Pinteon Therapeutics, Prothena, Quanterix, Red Abbey Labs, reMYND, Roche, Samumed, ScandiBio Therapeutics AB, Siemens Healthineers, Triplet Therapeutics, and Wave, has given lectures sponsored by Alzecure, BioArctic, Biogen, Cellectricon, Fujirebio, LabCorp, Lilly, Novo Nordisk, Oy Medix Biochemica AB, Roche, and WebMD, is a co-founder of Brain Biomarker Solutions in Gothenburg AB (BBS), which is a part of the GU Ventures Incubator Program, and is a shareholder of MicThera (outside submitted work).FJM, AD, IS, CL, GB, UA, and HK have no competing interests to declare that are relevant to the content of this article.","order":2,"name":"Ethics","group":{"name":"EthicsHeading","label":"Conflict of interests"}},{"value":"All participants provided written informed consent at enrollment including consent to publication. In the Gothenburg cohort, the H70 Clinical Studies was approved by the Ethical Review Authority in Sweden (ID: 2019\u201304024) and the Gothenburg H70 Birth Cohort Study was approved by the Regional Ethical Review Board in Gothenburg, Sweden (ID: 869\u201313). In the UCL ION cohort, dementia participants provided informed written consent (ethical permit 12\/LO\/1504). Ethical approval for HD patients was granted by the London Camberwell St Giles Ethics Committee (ref: 186932), for MS patients by the Scotland A Research Ethics Committee (REC: 13\/SS\/0007), and for healthy controls by the Hampstead Ethics Committee (ref: 19\/LO\/0913). Animal use for antibody production was approved by the Gothenburg's Animal Experimentation Ethics Committee (ID: 003795). This study complies with the Declaration of Helsinki.","order":3,"name":"Ethics","group":{"name":"EthicsHeading","label":"Ethical approval"}}],"article-number":"676"}}