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. 2025 May;21(5):e14621.
doi: 10.1002/alz.14621.

Biomarker discovery in Alzheimer's and neurodegenerative diseases using Nucleic Acid Linked Immuno-Sandwich Assay

Nicholas J Ashton  1   2   3 , Andrea L Benedet  1 , Guglielmo Di Molfetta  1 , Ilaria Pola  1 , Federica Anastasi  4   5 , Aida Fernández-Lebrero  4   5   6   7 , Albert Puig-Pijoan  4   6   8 , Ashvini Keshavan  9 , Jonathan Schott  9   10 , Kubra Tan  1 , Joel Simrén  1 , Bárbara Fernandes Gomes  1 , Laia Montoliu-Gaya  1 , Richard Isaacson  11   12 , Matilde Bongianni  13 , Chiara Tolassi  14 , Valentina Cantoni  15 , Antonella Alberici  15 , Alessandro Padovani  15   16   17   18 , Gianluigi Zanusso  19 , Andrea Pilotto  15   16   17   18 , Barbara Borroni  15   16 , Marc Suárez-Calvet  4   5   6   20 , Kaj Blennow  1   21   22   23 , Henrik Zetterberg  1   9   10   21   24   25
Affiliations

Biomarker discovery in Alzheimer's and neurodegenerative diseases using Nucleic Acid Linked Immuno-Sandwich Assay

Nicholas J Ashton et al. Alzheimers Dement. 2025 May.

Abstract

Introduction: Recent advancements in immunological methods accurately quantify biofluid biomarkers for Alzheimer's disease (AD) pathology. Despite progress, more biomarkers, ideally in blood, are needed for effective disease monitoring for AD and other neurodegenerative proteinopathies.

Methods: We used the Nucleic Acid Linked Immuno-Sandwich Assay (NULISA) central nervous system panel for biomarker quantification in plasma, serum, and cerebrospinal fluid of patients with AD, mild cognitive impairment, Lewy body dementia, progranulin (GRN) mutation carriers.

Results: NULISA identified phosphorylated tau217 and neurofilament light chain as the most deregulated biomarkers in the AD continuum and GRN mutation carriers, respectively. Importantly, numerous novel proteomic changes were observed in each disease endophenotype, which included synaptic processing, inflammation, microglial reactivity, TAR DNA-binding protein 43, and α-synuclein pathology.

Discussion: We underline the potential of next-generation biomarker identification tools to detect novel proteomic features that also incorporate established biomarkers. These findings highlight the importance of continued biomarker discovery to improve treatment decisions and help us better understand the complexities of neurodegenerative disorders.

Highlights: The, direct, or indirect, measures in blood that complement phosphorylated tau (p-tau)217 for other proteinopathies or disease progression are urgently needed. Significant novel proteomic changes were observed in each disease endophenotype in plasma, serum, and cerebrospinal fluid, which included proteins involved in synaptic processing, inflammation, microglial reactivity, TAR DNA-binding protein 43, and α-synuclein pathology. Nucleic Acid Linked Immuno-Sandwich Assay continued to unbiasely highlight p-tau217 and neurofilament light chain as the most significantly deregulated blood biomarkers in the Alzheimer's disease continuum and progranulin mutation carriers, respectively.

Keywords: Lewy body disease; Nucleic Acid Linked Immuno‐Sandwich Assay; discovery; frontotemporal dementia; plasma biomarkers; proteomics.

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Conflict of interest statement

HZ has served on 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, LabCorp, Merry Life, Nervgen, Novo Nordisk, Optoceutics, Passage Bio, Pinteon Therapeutics, Prothena, Red Abbey Labs, reMYND, Roche, Samumed, Siemens Healthineers, Triplet Therapeutics, and Wave, has given lectures in symposia sponsored by Alzecure, Biogen, Cellectricon, Fujirebio, Lilly, Novo Nordisk, and Roche, and is a co‐founder of Brain Biomarker Solutions in Gothenburg AB (BBS), which is a part of the GU Ventures Incubator Program (outside submitted work). BB has served on scientific advisory boards for Alector, Alexion, AviadoBio, Denali, Lilly/Prevail, UCB, and Wave. APi received consultancy/speaker fees from Abbvie, Bial, Lundbeck, Roche, and Zambon pharmaceuticals. APa received grant support from Ministry of Health (MINSAL) and Ministry of Education, Research and University (MIUR), from CARIPLO Foundation; personal compensation as a consultant/scientific advisory board member for Biogen, Lundbeck, Roche, Nutricia, General Healthcare (GE). MS‐C has given lectures in symposia sponsored by Almirall, Eli Lilly, Novo Nordisk, Roche Diagnostics, and Roche Farma; received consultancy fees (paid to the institution) from Roche Diagnostics; and served on advisory boards of Eli Lilly, Grifols, and Roche Diagnostics. He was granted a project and is a site investigator of a clinical trial (funded to the institution) by Roche Diagnostics. In‐kind support for research (to the institution) was received from ADx Neurosciences, Alamar Biosciences, Avid Radiopharmaceuticals, Eli Lilly, Fujirebio, Janssen Research & Development, and Roche Diagnostics. All other authors do not report any conflicts of interest. Author disclosures are available in the supporting information.

Figures

FIGURE 1
FIGURE 1
Differential protein changes between biologically determined AD compared to non‐AD (cohort 1) in plasma (A), serum (B), and CSF (C). AD, Alzheimer's disease; CSF, cerebrospinal fluid; FC, fold change.
FIGURE 2
FIGURE 2
MCI due to AD patients and MCI unlikely due to AD (A), LB+ compared to LB– (B), and GRN mutation carriers compared to non‐carriers (C). AD, Alzheimer's disease; FC, fold change; GRN, progranulin; LB, Lewy body; MCI, mild cognitive impairment.
See this image and copyright information in PMC

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