Ultrasensitive Protein Aggregate Quantification Assays for Neurodegenerative Diseases on the Simoa Platform
Nanoscale aggregates play a key role in the pathogenesis of neurodegenerative disorders such as Alzheimer's and Parkinson's disease. However, quantifying these aggregates in complex biological samples, such as biofluids and postmortem brain tissue, has been challenging due to their low concentration and small size, necessitating the development of methods with high sensitivity and specificity. Here, we have developed ultrasensitive assays utilizing the Quanterix Simoa platform to detect α-synuclein, β-amyloid and tau aggregates, including those with common posttranslational modifications such as truncation of α-synuclein and AT8 phosphorylation of tau aggregates. All assays had a detection limit in the low pM range. As a part of this work, we developed silica-nanoparticle calibrators, allowing for the quantification of all aggregates. These assays were validated for aggregate and target specificity through denaturation and cross-reactivity experiments. We then applied these assays to brain homogenate samples from Alzheimer's disease and control samples, demonstrating their applicability to postmortem tissue. Lastly, we explored the potential of these assays for blood-based diagnostics by detecting aggregates in serum samples from early Alzheimer's disease patients.
Authors
- Emily P. English (ORCID: https://orcid.org/0009-0000-1621-4669)
- Emre Fertan (ORCID: https://orcid.org/0000-0002-0060-5806)
- David A. Klenerman (ORCID: https://orcid.org/0000-0001-7116-6954)
- Jeff Y. L. Lam (ORCID: https://orcid.org/0000-0001-7408-2342)
- Zengjie Xia (ORCID: https://orcid.org/0000-0002-5977-9691)
- Gonçalo J. L. Bernardes (ORCID: https://orcid.org/0000-0001-6594-8917)
- Matthew R. Cheetham (ORCID: https://orcid.org/0000-0001-7412-2332)
- Juraj Konč (ORCID: https://orcid.org/0000-0001-8681-2082)
- Dorothea Böken (ORCID: https://orcid.org/0009-0008-8443-4469)
- Henrik Zetterberg (ORCID: https://orcid.org/0000-0003-3930-4354)
- Florence Layburn
- Yunzhao Wu
Institutions
- University of Cambridge (GB)
- Sahlgrenska University Hospital (SE)
- UK Dementia Research Institute (GB)
- MRC Prion Unit (GB)
- Universitas Islam Syekh Yusuf (ID)
- University College London (GB)
- University of Gothenburg (SE)
Publication Details
- Journal
- Analytical Chemistry
- Published
- 2024-12-24
- DOI
- https://doi.org/10.1021/acs.analchem.4c04188
- Citations
- 16
- Primary Topic
- Alzheimer's disease research and treatments
- Type
- article
- Field-Weighted Citation Impact
- 5.22
Funders
- University of Wisconsin-Madison
- Alzheimer's Society
- Alzheimer Society
- National Institute for Health and Care Research
- Royal Society
- Alzheimer’s Research UK
- Sahlgrenska Universitetssjukhuset
- Göteborgs Universitet
- UK Dementia Research Institute
- School of Medicine and Public Health, University of Wisconsin-Madison
- Medical Research Council
- Sahlgrenska Akademin