Comparative Analysis of Total Alpha-Synuclein (αSYN) Immunoassays Reveals That They Do Not Capture the Diversity of Modified αSYN Proteoforms.
| Title: | Comparative Analysis of Total Alpha-Synuclein (αSYN) Immunoassays Reveals That They Do Not Capture the Diversity of Modified αSYN Proteoforms. |
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| Authors: | Petricca L; ND Biosciences SA, Epalinges, Switzerland.; Chiki N; ND Biosciences SA, Epalinges, Switzerland.; Hanna-El-Daher L; Laboratory of Molecular and Chemical Biology of Neurodegeneration, Brain Mind Institute,Ecole Polytechnique Fédérale de Lausanne (EPFL), Lausanne, Switzerland.; Aeschbach L; Laboratory of Molecular and Chemical Biology of Neurodegeneration, Brain Mind Institute,Ecole Polytechnique Fédérale de Lausanne (EPFL), Lausanne, Switzerland.; Burai R; Laboratory of Molecular and Chemical Biology of Neurodegeneration, Brain Mind Institute,Ecole Polytechnique Fédérale de Lausanne (EPFL), Lausanne, Switzerland.; Stoops E; ADx NeuroSciences NV, Technologiepark 94 - Bio Incubator, Gent, Belgium.; Fares MB; ND Biosciences SA, Epalinges, Switzerland.; Lashuel HA; ND Biosciences SA, Epalinges, Switzerland.; Laboratory of Molecular and Chemical Biology of Neurodegeneration, Brain Mind Institute,Ecole Polytechnique Fédérale de Lausanne (EPFL), Lausanne, Switzerland. |
| Source: | Journal of Parkinson's disease [J Parkinsons Dis] 2022; Vol. 12 (5), pp. 1449-1462. |
| Publication Type: | Journal Article; Research Support, Non-U.S. Gov't |
| Language: | English |
| Journal Info: | Publisher: SAGE Publications Country of Publication: United States NLM ID: 101567362 Publication Model: Print Cited Medium: Internet ISSN: 1877-718X (Electronic) Linking ISSN: 18777171 NLM ISO Abbreviation: J Parkinsons Dis Subsets: MEDLINE |
| Imprint Name(s): | Publication: 2024- : [Thousand Oaks, CA] : SAGE Publications; Original Publication: Amsterdam : IOS Press |
| MeSH Terms: | Parkinson Disease*/diagnosis ; alpha-Synuclein*/metabolism; Biomarkers ; Humans ; Immunoassay |
| Abstract: | Background: The development of therapeutics for Parkinson's disease (PD) requires the establishment of biomarker assays to enable stratifying patients, monitoring disease progression, and assessing target engagement. Attempts to develop diagnostic assays based on detecting levels of the α-synuclein (αSYN) protein, a central player in the pathogenesis of PD, have yielded inconsistent results.; Objective: To determine whether the three commercial kits that have been extensively used for total αSYN quantification in human biological fluids (from Euroimmun, MSD, and Biolegend) are capable of capturing the diversity and complexity of relevant αSYN proteoforms.; Methods: We investigated and compared the ability of the different assays to detect the diversity of αSYN proteoforms using a library of αSYN proteins that comprise the majority of disease-relevant αSYN variants and post-translational modifications (PTMs).; Results: Our findings showed that none of the three tested immunoassays accurately capture the totality of relevant αSYN species, and that these assays are unable to recognize most disease-associated C-terminally truncated variants of αSYN. Moreover, several N-terminal truncations and phosphorylation/nitration PTMs differentially modify the level of αSYN detection and recovery by different immunoassays, and a CSF matrix effect was observed for most of the αSYN proteoforms analyzed by the three immunoassays.; Conclusion: Our results show that the tested immunoassays do not capture the totality of the relevant αSYN species and therefore may not be appropriate tools to provide an accurate measure of total αSYN levels in samples containing modified forms of the protein. This highlights the need for next generation αSYN immunoassays that capture the diversity of αSYN proteoforms. |
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| Contributed Indexing: | Keywords: Alpha-synuclein; ELISA; Parkinson’s disease; antibodies; cerebrospinal fluid; immunoassays; post-translational modifications; truncations |
| Substance Nomenclature: | 0 (Biomarkers); 0 (alpha-Synuclein) |
| Entry Date(s): | Date Created: 20220509 Date Completed: 20220712 Latest Revision: 20220917 |
| Update Code: | 20260130 |
| PubMed Central ID: | PMC9398082 |
| DOI: | 10.3233/JPD-223285 |
| PMID: | 35527570 |
| Database: | MEDLINE |
Journal Article; Research Support, Non-U.S. Gov't