Analytical Methods for Fluid Biomarkers in Alzheimer’s Disease from Discovery to Clinical Implementation
- 1Department of Biomedicine, Neurosciences, and Advanced Diagnostics, Institute of Clinical Biochemistry, Clinical Molecular Medicine, and Clinical Laboratory Medicine, University of Palermo, 90127 Palermo, Italy
- 2Department of Laboratory Medicine, University Hospital Paolo Giaccone, 90127 Palermo, Italy
- 3University Laboratory of Clinical Pathology and Toxicology, Pio XI Hospital, Desio, ASST Brianza, 20832 Desio, Italy
Abstract
Alzheimer's disease (AD) is increasingly recognized as a biological continuum characterized by early neuropathological and molecular changes that precede the onset of clinical symptoms. Fluid biomarkers have transformed the diagnostic landscape by enabling the in vivo detection of core AD pathologies, particularly amyloid-β deposition and tau-related neurodegeneration. Despite the rapid expansion of candidate biomarkers, however, only a limited number have successfully translated into clinical practice. Discovery-phase approaches, primarily driven by mass spectrometry-based proteomics, enable the unbiased identification of novel biomarker candidates across multiple biological pathways. Research-phase methods, including immunoassays such as enzyme-linked immunosorbent assay (ELISA), electrochemiluminescence immunoassays (ECLIA), microfluidic platforms, and ultrasensitive technologies such as single-molecule array (SIMOA), support analytical and clinical validation in well-characterized cohorts. Clinical implementation has been advanced by fully automated platforms, including Lumipulse and Elecsys, which have obtained regulatory approval for cerebrospinal fluid biomarkers and, more recently, blood-based biomarkers. These developments represent a paradigm shift toward minimally invasive and scalable diagnostic strategies that may reduce dependence on neuroimaging techniques. Nevertheless, major challenges remain, including assay standardization, inter-platform variability, demonstration of clinical utility, and barriers to widespread clinical adoption. This review provides a comprehensive overview of analytical methods used to measure AD fluid biomarkers in cerebrospinal fluid and plasma, structured according to the biomarker development pipeline from discovery to clinical implementation. Overall, the review highlights a fit-for-purpose approach to biomarker development and emphasizes the complementary roles of diverse analytical technologies across the different phases of biomarker translation.
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BibTeX
@article{agnello2026analytical,
title = {{Analytical Methods for Fluid Biomarkers in Alzheimer’s Disease from Discovery to Clinical Implementation}},
author = {Luisa Agnello and Roberto Dominici and Caterina Maria Gambino and Concetta Scazzone and Marcello Ciaccio},
journal = {International Journal of Molecular Sciences},
year = {2026},
month = may,
doi = {10.3390/ijms27104518},
url = {https://doi.org/10.3390/ijms27104518},
note = {CC BY},
}
RIS
TY - JOUR
TI - Analytical Methods for Fluid Biomarkers in Alzheimer’s Disease from Discovery to Clinical Implementation
AU - Luisa Agnello
AU - Roberto Dominici
AU - Caterina Maria Gambino
AU - Concetta Scazzone
AU - Marcello Ciaccio
PY - 2026
DA - 2026/05/18
JO - International Journal of Molecular Sciences
DO - 10.3390/ijms27104518
UR - https://doi.org/10.3390/ijms27104518
AB - Alzheimer's disease (AD) is increasingly recognized as a biological continuum characterized by early neuropathological and molecular changes that precede the onset of clinical symptoms. Fluid biomarkers have transformed the diagnostic landscape by enabling the in vivo detection of core AD pathologies, particularly amyloid-β deposition and tau-related neurodegeneration. Despite the rapid expansion of candidate biomarkers, however, only a limited number have successfully translated into clinical practice. Discovery-phase approaches, primarily driven by mass spectrometry-based proteomics, enable the unbiased identification of novel biomarker candidates across multiple biological pathways. Research-phase methods, including immunoassays such as enzyme-linked immunosorbent assay (ELISA), electrochemiluminescence immunoassays (ECLIA), microfluidic platforms, and ultrasensitive technologies such as single-molecule array (SIMOA), support analytical and clinical validation in well-characterized cohorts. Clinical implementation has been advanced by fully automated platforms, including Lumipulse and Elecsys, which have obtained regulatory approval for cerebrospinal fluid biomarkers and, more recently, blood-based biomarkers. These developments represent a paradigm shift toward minimally invasive and scalable diagnostic strategies that may reduce dependence on neuroimaging techniques. Nevertheless, major challenges remain, including assay standardization, inter-platform variability, demonstration of clinical utility, and barriers to widespread clinical adoption. This review provides a comprehensive overview of analytical methods used to measure AD fluid biomarkers in cerebrospinal fluid and plasma, structured according to the biomarker development pipeline from discovery to clinical implementation. Overall, the review highlights a fit-for-purpose approach to biomarker development and emphasizes the complementary roles of diverse analytical technologies across the different phases of biomarker translation.
SN - 1422-0067
SN - 1661-6596
ER -