{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,8,18]],"date-time":"2026-08-18T15:53:58Z","timestamp":1787068438191,"version":"build-2736575974"},"reference-count":28,"publisher":"MDPI AG","issue":"12","license":[{"start":{"date-parts":[[2024,11,28]],"date-time":"2024-11-28T00:00:00Z","timestamp":1732752000000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"name":"Target ALS","award":["TT-2019-IMA-S1"],"award-info":[{"award-number":["TT-2019-IMA-S1"]}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Biosensors"],"abstract":"<jats:p>Transactive response DNA-binding protein of 43 kDa (TDP-43) is a major component of pathological inclusions in various neurodegenerative disorders, including amyotrophic lateral sclerosis and frontotemporal lobar degeneration. The detection of TDP-43 in biofluids is crucial for the development of diagnostic and prognostic indicators of disease and therapeutic development for TDP-43-related proteinopathies. Despite its potential as a biomarker for numerous neurological disorders, the lack of a sensitive and reproducible TDP-43 assay hinders progress in TDP-43-based therapy development, underscoring the need for an effective and standardized method for accurate quantification. Addressing the limitations of sensitivity and reproducibility in existing assays, in this study, we developed and validated a highly sensitive electrochemiluminescence immunoassay on the Meso Scale Discovery platform. The assay demonstrated the detection of full-length TDP-43 in human biofluids with a limit of detection of 4pg\/mL, a working range of 4\u201320,000 pg\/mL, and a total assay time of 16 h. In this study, we developed and validated a sensitive immunoassay for the detection of full-length TDP-43 in human biofluids using the Meso Scale Discovery platform. We used this immunoassay to quantify TDP-43 levels in the plasma and serum of healthy controls and ALS patients. Our results indicate a reduction in full-length TDP-43 in the blood of ALS patients compared to healthy controls.<\/jats:p>","DOI":"10.3390\/bios14120578","type":"journal-article","created":{"date-parts":[[2024,11,28]],"date-time":"2024-11-28T08:15:54Z","timestamp":1732781754000},"page":"578","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":4,"title":["Development of a Sensitive and Reliable Meso Scale Discovery-Based Electrochemiluminescence Immunoassay to Quantify TDP-43 in Human Biofluids"],"prefix":"10.3390","volume":"14","author":[{"given":"Jiyan","family":"An","sequence":"first","affiliation":[{"name":"Department of Translational Neuroscience, Barrow Neurological Institute, Phoenix, AZ 85013, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-1746-7315","authenticated-orcid":false,"given":"Lathika","family":"Gopalakrishnan","sequence":"additional","affiliation":[{"name":"Department of Translational Neuroscience, Barrow Neurological Institute, Phoenix, AZ 85013, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-7079-7735","authenticated-orcid":false,"given":"Vanessa","family":"Ortega","sequence":"additional","affiliation":[{"name":"Department of Translational Neuroscience, Barrow Neurological Institute, Phoenix, AZ 85013, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Justin","family":"Saul","sequence":"additional","affiliation":[{"name":"Department of Translational Neuroscience, Barrow Neurological Institute, Phoenix, AZ 85013, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Renu","family":"Kadali","sequence":"additional","affiliation":[{"name":"Department of Translational Neuroscience, Barrow Neurological Institute, Phoenix, AZ 85013, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-5404-7259","authenticated-orcid":false,"given":"Robert","family":"Bowser","sequence":"additional","affiliation":[{"name":"Department of Translational Neuroscience, Barrow Neurological Institute, Phoenix, AZ 85013, USA"},{"name":"Department of Neurology, Barrow Neurological Institute, Phoenix, AZ 85013, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2024,11,28]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"1182","DOI":"10.1016\/S1474-4422(16)30199-5","article-title":"Amyotrophic lateral sclerosis: Moving towards a new classification system","volume":"15","author":"Hardiman","year":"2016","journal-title":"Lancet Neurol."},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"1503","DOI":"10.1093\/brain\/awz099","article-title":"Limbic-predominant age-related TDP-43 encephalopathy (LATE): Consensus working group report","volume":"142","author":"Nelson","year":"2019","journal-title":"Brain"},{"key":"ref_3","doi-asserted-by":"crossref","first-page":"26","DOI":"10.1016\/j.clinbiochem.2019.08.005","article-title":"Alzheimer\u2019s disease and other neurodegenerative dementias in comorbidity: A clinical and neuropathological overview","volume":"73","author":"Matej","year":"2019","journal-title":"Clin. Biochem."},{"key":"ref_4","doi-asserted-by":"crossref","first-page":"3778","DOI":"10.1242\/jcs.038950","article-title":"Structural determinants of the cellular localization and shuttling of TDP-43","volume":"121","author":"Ayala","year":"2008","journal-title":"J. Cell Sci."},{"key":"ref_5","doi-asserted-by":"crossref","first-page":"13302","DOI":"10.1074\/jbc.M800342200","article-title":"Disturbance of nuclear and cytoplasmic TAR DNA-binding protein (TDP-43) induces disease-like redistribution, sequestration, and aggregate formation","volume":"283","author":"Winton","year":"2008","journal-title":"J. Biol. Chem."},{"key":"ref_6","doi-asserted-by":"crossref","first-page":"427","DOI":"10.1002\/ana.21147","article-title":"Pathological TDP-43 distinguishes sporadic amyotrophic lateral sclerosis from amyotrophic lateral sclerosis with SOD1 mutations","volume":"61","author":"Mackenzie","year":"2007","journal-title":"Ann. Neurol."},{"key":"ref_7","doi-asserted-by":"crossref","first-page":"36337","DOI":"10.1074\/jbc.M104236200","article-title":"Characterization and functional implications of the RNA binding properties of nuclear factor TDP-43, a novel splicing regulator of CFTR exon 9","volume":"276","author":"Buratti","year":"2001","journal-title":"J. Biol. Chem."},{"key":"ref_8","doi-asserted-by":"crossref","unstructured":"Prasad, A., Bharathi, V., Sivalingam, V., Girdhar, A., and Patel, B.K. (2019). Molecular Mechanisms of TDP-43 Misfolding and Pathology in Amyotrophic Lateral Sclerosis. Front. Mol. Neurosci., 12.","DOI":"10.3389\/fnmol.2019.00025"},{"key":"ref_9","doi-asserted-by":"crossref","first-page":"7","DOI":"10.1111\/jnc.14327","article-title":"Pathomechanisms of TDP-43 in neurodegeneration","volume":"146","author":"Gao","year":"2018","journal-title":"J. Neurochem."},{"key":"ref_10","doi-asserted-by":"crossref","unstructured":"Dyer, M.S., Woodhouse, A., and Blizzard, C.A. (2021). Cytoplasmic Human TDP-43 Mislocalization Induces Widespread Dendritic Spine Loss in Mouse Upper Motor Neurons. Brain Sci., 11.","DOI":"10.3390\/brainsci11070883"},{"key":"ref_11","doi-asserted-by":"crossref","first-page":"3187","DOI":"10.1093\/brain\/aww237","article-title":"Exosome secretion is a key pathway for clearance of pathological TDP-43","volume":"139","author":"Iguchi","year":"2016","journal-title":"Brain"},{"key":"ref_12","doi-asserted-by":"crossref","unstructured":"Zhang, N., Gu, D., Meng, M., and Gordon, M.L. (2020). TDP-43 Is Elevated in Plasma Neuronal-Derived Exosomes of Patients with Alzheimer\u2019s Disease. Front. Aging Neurosci., 12.","DOI":"10.3389\/fnagi.2020.00166"},{"key":"ref_13","doi-asserted-by":"crossref","first-page":"55","DOI":"10.1007\/s00401-008-0456-1","article-title":"Increased TDP-43 protein in cerebrospinal fluid of patients with amyotrophic lateral sclerosis","volume":"117","author":"Kasai","year":"2009","journal-title":"Acta Neuropathol."},{"key":"ref_14","doi-asserted-by":"crossref","first-page":"1481","DOI":"10.1001\/archneur.65.11.1481","article-title":"TDP-43 in cerebrospinal fluid of patients with frontotemporal lobar degeneration and amyotrophic lateral sclerosis","volume":"65","author":"Steinacker","year":"2008","journal-title":"Arch. Neurol."},{"key":"ref_15","doi-asserted-by":"crossref","unstructured":"Han, Z., Christiansen, A., and Liau, G. (2021). Relative Quantification of Na(V)1.1 Protein in Mouse Brains Using a Meso Scale Discovery-Electrochemiluminescence (MSD-ECL) Method. Bio Protoc., 11.","DOI":"10.21769\/BioProtoc.3910"},{"key":"ref_16","doi-asserted-by":"crossref","unstructured":"Kivisakk, P., Carlyle, B.C., Sweeney, T., Trombetta, B.A., LaCasse, K., El-Mufti, L., Tuncali, I., Chibnik, L.B., Das, S., and Scherzer, C.R. (2023). Plasma biomarkers for diagnosis of Alzheimer\u2019s disease and prediction of cognitive decline in individuals with mild cognitive impairment. Front. Neurol., 14.","DOI":"10.3389\/fneur.2023.1069411"},{"key":"ref_17","doi-asserted-by":"crossref","first-page":"67","DOI":"10.3233\/JAD-130203","article-title":"The AD-CSF-index discriminates Alzheimer\u2019s disease patients from healthy controls: A validation study","volume":"36","author":"Molinuevo","year":"2013","journal-title":"J. Alzheimers Dis."},{"key":"ref_18","doi-asserted-by":"crossref","unstructured":"Reindl, W., Baldo, B., Schulz, J., Janack, I., Lindner, I., Kleinschmidt, M., Sedaghat, Y., Thiede, C., Tillack, K., and Schmidt, C. (2019). Meso scale discovery-based assays for the detection of aggregated huntingtin. PLoS ONE, 14.","DOI":"10.1371\/journal.pone.0213521"},{"key":"ref_19","doi-asserted-by":"crossref","first-page":"659","DOI":"10.1016\/j.molmed.2011.06.004","article-title":"TDP-43 functions and pathogenic mechanisms implicated in TDP-43 proteinopathies","volume":"17","author":"Cohen","year":"2011","journal-title":"Trends Mol. Med."},{"key":"ref_20","doi-asserted-by":"crossref","first-page":"6330","DOI":"10.1007\/s12035-023-03472-y","article-title":"The Molecular Link Between TDP-43, Endogenous Retroviruses and Inflammatory Neurodegeneration in Amyotrophic Lateral Sclerosis: A Potential Target for Triumeq, an Antiretroviral Therapy","volume":"60","author":"Dubowsky","year":"2023","journal-title":"Mol. Neurobiol."},{"key":"ref_21","doi-asserted-by":"crossref","first-page":"641","DOI":"10.1007\/s10571-009-9489-9","article-title":"TDP-43 dimerizes in human cells in culture","volume":"30","author":"Shiina","year":"2010","journal-title":"Cell Mol. Neurobiol."},{"key":"ref_22","doi-asserted-by":"crossref","first-page":"1159","DOI":"10.1097\/NEN.0b013e31818e8951","article-title":"Colocalization of transactivation-responsive DNA-binding protein 43 and huntingtin in inclusions of Huntington disease","volume":"67","author":"Schwab","year":"2008","journal-title":"J. Neuropathol. Exp. Neurol."},{"key":"ref_23","doi-asserted-by":"crossref","first-page":"e24672","DOI":"10.1016\/j.heliyon.2024.e24672","article-title":"Validation of a newly developed immunoassay for TDP-43 in human plasma","volume":"10","author":"Matsuura","year":"2024","journal-title":"Heliyon"},{"key":"ref_24","doi-asserted-by":"crossref","first-page":"573","DOI":"10.1055\/s-0040-1714314","article-title":"Association of Plasma Biomarkers for Angiogenesis and Proteinopathy in Indian Amyotrophic Lateral Sclerosis Patients","volume":"11","author":"Modgil","year":"2020","journal-title":"J. Neurosci. Rural. Pract."},{"key":"ref_25","doi-asserted-by":"crossref","first-page":"151","DOI":"10.1186\/s13195-022-01091-8","article-title":"Serum total TDP-43 levels are decreased in frontotemporal dementia patients with C9orf72 repeat expansion or concomitant motoneuron disease phenotype","volume":"14","author":"Katisko","year":"2022","journal-title":"Alzheimers Res. Ther."},{"key":"ref_26","doi-asserted-by":"crossref","unstructured":"Berning, B.A., and Walker, A.K. (2019). The Pathobiology of TDP-43 C-Terminal Fragments in ALS and FTLD. Front. Neurosci., 13.","DOI":"10.3389\/fnins.2019.00335"},{"key":"ref_27","doi-asserted-by":"crossref","first-page":"41","DOI":"10.1016\/j.neuint.2018.12.009","article-title":"TDP-43 levels are higher in platelets from patients with sporadic amyotrophic lateral sclerosis than in healthy controls","volume":"124","author":"Hishizawa","year":"2019","journal-title":"Neurochem. Int."},{"key":"ref_28","doi-asserted-by":"crossref","unstructured":"Luthi-Carter, R., Cappelli, S., Le Roux-Bourdieu, M., Tentillier, N., Quinn, J.P., Petrozziello, T., Gopalakrishnan, L., Sethi, P., Choudhary, H., and Bartolini, G. (2024). Location and function of TDP-43 in platelets, alterations in neurodegenerative diseases and arising considerations for current plasma biobank protocols. Sci. Rep., 14.","DOI":"10.1038\/s41598-024-77057-7"}],"container-title":["Biosensors"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/2079-6374\/14\/12\/578\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,10,10]],"date-time":"2025-10-10T16:41:46Z","timestamp":1760114506000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/2079-6374\/14\/12\/578"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2024,11,28]]},"references-count":28,"journal-issue":{"issue":"12","published-online":{"date-parts":[[2024,12]]}},"alternative-id":["bios14120578"],"URL":"https:\/\/doi.org\/10.3390\/bios14120578","relation":{},"ISSN":["2079-6374"],"issn-type":[{"value":"2079-6374","type":"electronic"}],"subject":[],"published":{"date-parts":[[2024,11,28]]}}}