Advertisement

Analytical and Bioanalytical Chemistry

, Volume 411, Issue 1, pp 267–275 | Cite as

Development of an automated capillary nano-immunoassay—Simple Western assay—to quantify total TDP43 protein in human platelet samples

  • Anthony FourierEmail author
  • Jean Escal
  • Emilien Bernard
  • Ingolf Lachman
  • Armand Perret-Liaudet
  • Pascal Leblanc
  • Isabelle Quadrio
Research Paper

Abstract

Frontotemporal lobar degeneration syndrome is the second cause of young-onset dementia. Unfortunately, reliable biomarkers are currently lacking for the diagnosis of this disease. As TDP43 protein is one of the proteins pathologically involved in frontotemporal lobar degeneration, many studies have been performed to assess TDP43 protein diagnostic performances. Mixed results were obtained using cerebrospinal fluid and plasma samples so far. The aim of the study was to develop an automated capillary nano-immunoassay—Simple Western assay—to detect and quantify TDP43 protein simultaneously in human blood-based samples. Simple Western assay was developed with two different cell lysates used as positive controls and was compared to Western blot. TDP43 protein profiles in plasma samples were disappointing, as they were discordant to our positive controls. On the contrary, similar TDP43 patterns were obtained between platelet samples and cell lysates using both assays. Simple Western assay provided good quantitative performances in platelet samples: a linearity of signals could be observed (r2 = 0.994), associated to a within-run variability at 5.7%. Preliminary results based on a cohort of patients suffering from frontotemporal lobar degeneration showed large inter-individual variations superior to Simple Western’s analytical variability. Simple Western assay seems to be suitable for detecting and quantifying TDP43 protein in platelet samples, providing a potential candidate biomarker in this disease. Further confirmation studies should now be performed on larger cohorts of patients to assess diagnostic performances of TDP43 protein in platelet samples.

Keywords

Dementia TDP43 proteinopathies Capillary electrophoresis/electrophoresis High-throughput screening assays Biomarkers 

Notes

Compliance with ethical standards

Conflict of interest

Ingolf Lachmann provided 2G10 antibody and reports that he is employee of AJ Roboscreen GmbH®, Leipzig, Germany. No other conflict of interest is reported.

Ethical approval

This study was based on biological samples collected for routine diagnosis in the framework of a medical follow-up recommended for FTLD patients. The samples associated to written consents were stored in a bio bank with the authorization from the French Ministry of Health (Declaration Number DC-2008-304). Authorization for handling personal data was granted by the French data protection commission [Commission Nationale de l’Informatique et des Libertés (CNIL)].

The patients included in this study or their legal next of kin gave two informed written consents: the first one for FTLD genetic testing and the second one authorizing a change of biological purpose for remaining parts of samples.

References

  1. 1.
    Rabinovici GD, Miller BL. Frontotemporal lobar degeneration: epidemiology, pathophysiology, diagnosis and management. CNS Drugs. 2010;24(5):375–98.  https://doi.org/10.2165/11533100-000000000-00000.CrossRefPubMedPubMedCentralGoogle Scholar
  2. 2.
    Warren JD, Rohrer JD, Rossor MN. Clinical review. Frontotemporal dementia. BMJ. 2013;347:f4827.  https://doi.org/10.1136/bmj.f4827.CrossRefPubMedPubMedCentralGoogle Scholar
  3. 3.
    Neary D, Snowden J, Mann D. Frontotemporal dementia. Lancet Neurol. 2005;4(11):771–80.  https://doi.org/10.1016/S1474-4422(05)70223-4.CrossRefPubMedGoogle Scholar
  4. 4.
    Grossman M. Primary progressive aphasia: clinicopathological correlations. Nat Rev Neurol. 2010;6(2):88–97.  https://doi.org/10.1038/nrneurol.2009.216.CrossRefPubMedPubMedCentralGoogle Scholar
  5. 5.
    Irwin DJ, Cairns NJ, Grossman M, McMillan CT, Lee EB, Van Deerlin VM, et al. Frontotemporal lobar degeneration: defining phenotypic diversity through personalized medicine. Acta Neuropathol. 2015;129(4):469–91.  https://doi.org/10.1007/s00401-014-1380-1.CrossRefPubMedGoogle Scholar
  6. 6.
    Rascovsky K, Hodges JR, Knopman D, Mendez MF, Kramer JH, Neuhaus J, et al. Sensitivity of revised diagnostic criteria for the behavioural variant of frontotemporal dementia. Brain. 2011;134(Pt 9):2456–77.  https://doi.org/10.1093/brain/awr179. CrossRefPubMedPubMedCentralGoogle Scholar
  7. 7.
    Gorno-Tempini ML, Hillis AE, Weintraub S, Kertesz A, Mendez M, Cappa SF, et al. Classification of primary progressive aphasia and its variants. Neurology. 2011;76(11):1006–14.  https://doi.org/10.1212/WNL.0b013e31821103e6.CrossRefPubMedPubMedCentralGoogle Scholar
  8. 8.
    Neumann M, Sampathu DM, Kwong LK, Truax AC, Micsenyi MC, Chou TT, et al. Ubiquitinated TDP-43 in frontotemporal lobar degeneration and amyotrophic lateral sclerosis. Science. 2006;314(5796):130–3.  https://doi.org/10.1126/science.1134108.CrossRefPubMedGoogle Scholar
  9. 9.
    Gao J, Wang L, Huntley ML, Perry G, Wang X. Pathomechanisms of TDP-43 in neurodegeneration. J Neurochem. 2018.  https://doi.org/10.1111/jnc.14327.
  10. 10.
    Buratti E. TDP-43 post-translational modifications in health and disease. Expert Opin Ther Targets. 2018;22(3):279–93.  https://doi.org/10.1080/14728222.2018.1439923.CrossRefPubMedGoogle Scholar
  11. 11.
    Geser F, Prvulovic D, O'Dwyer L, Hardiman O, Bede P, Bokde AL, et al. On the development of markers for pathological TDP-43 in amyotrophic lateral sclerosis with and without dementia. Prog Neurobiol. 2011;95(4):649–62.  https://doi.org/10.1016/j.pneurobio.2011.08.011.CrossRefPubMedPubMedCentralGoogle Scholar
  12. 12.
    McKhann GM, Knopman DS, Chertkow H, Hyman BT, Jack CR Jr, Kawas CH, et al. The diagnosis of dementia due to Alzheimer’s disease: recommendations from the National Institute on Aging-Alzheimer's Association workgroups on diagnostic guidelines for Alzheimer's disease. Alzheimers Dement. 2011;7(3):263–9.  https://doi.org/10.1016/j.jalz.2011.03.005.CrossRefPubMedPubMedCentralGoogle Scholar
  13. 13.
    Snyder HM, Carrillo MC, Grodstein F, Henriksen K, Jeromin A, Lovestone S, et al. Developing novel blood-based biomarkers for Alzheimer’s disease. Alzheimers Dement. 2014;10(1):109–14.  https://doi.org/10.1016/j.jalz.2013.10.007.CrossRefPubMedPubMedCentralGoogle Scholar
  14. 14.
    Riedl L, Mackenzie IR, Forstl H, Kurz A, Diehl-Schmid J. Frontotemporal lobar degeneration: current perspectives. Neuropsychiatr Dis Treat. 2014;10:297–310.  https://doi.org/10.2147/NDT.S38706.CrossRefPubMedPubMedCentralGoogle Scholar
  15. 15.
    Kasai T, Tokuda T, Ishigami N, Sasayama H, Foulds P, Mitchell DJ, et al. Increased TDP-43 protein in cerebrospinal fluid of patients with amyotrophic lateral sclerosis. Acta Neuropathol. 2009;117(1):55–62.  https://doi.org/10.1007/s00401-008-0456-1.CrossRefPubMedGoogle Scholar
  16. 16.
    Foulds PG, Davidson Y, Mishra M, Hobson DJ, Humphreys KM, Taylor M, et al. Plasma phosphorylated-TDP-43 protein levels correlate with brain pathology in frontotemporal lobar degeneration. Acta Neuropathol. 2009;118(5):647–58.  https://doi.org/10.1007/s00401-009-0594-0.CrossRefPubMedPubMedCentralGoogle Scholar
  17. 17.
    Feneberg E, Gray E, Ansorge O, Talbot K, Turner MR. Towards a TDP-43-based biomarker for ALS and FTLD. Mol Neurobiol. 2018.  https://doi.org/10.1007/s12035-018-0947-6.
  18. 18.
    Anderson GJ, C MC, Kennedy RT. Western blotting using capillary electrophoresis. Anal Chem. 2011;83(4):1350–5.  https://doi.org/10.1021/ac102671n.CrossRefPubMedPubMedCentralGoogle Scholar
  19. 19.
    Hughes AJ, Herr AE. Microfluidic Western blotting. Proc Natl Acad Sci U S A. 2012;109(52):21450–5.  https://doi.org/10.1073/pnas.1207754110. CrossRefPubMedPubMedCentralGoogle Scholar
  20. 20.
    Chen JQ, Wakefield LM, Goldstein DJ. Capillary nano-immunoassays: advancing quantitative proteomics analysis, biomarker assessment, and molecular diagnostics. J Transl Med. 2015;13:182.  https://doi.org/10.1186/s12967-015-0537-6.CrossRefPubMedPubMedCentralGoogle Scholar
  21. 21.
    Behari M, Shrivastava M. Role of platelets in neurodegenerative diseases: a universal pathophysiology. Int J Neurosci. 2013;123(5):287–99.  https://doi.org/10.3109/00207454.2012.751534.CrossRefPubMedGoogle Scholar
  22. 22.
    Wood EM, Falcone D, Suh E, Irwin DJ, Chen-Plotkin AS, Lee EB, et al. Development and validation of pedigree classification criteria for frontotemporal lobar degeneration. JAMA Neurol. 2013;70(11):1411–7.  https://doi.org/10.1001/jamaneurol.2013.3956.CrossRefPubMedPubMedCentralGoogle Scholar
  23. 23.
    Akimoto C, Volk AE, van Blitterswijk M, Van den Broeck M, Leblond CS, Lumbroso S, et al. A blinded international study on the reliability of genetic testing for GGGGCC-repeat expansions in C9orf72 reveals marked differences in results among 14 laboratories. J Med Genet. 2014;51(6):419–24.  https://doi.org/10.1136/jmedgenet-2014-102360.CrossRefPubMedPubMedCentralGoogle Scholar
  24. 24.
    Goossens J, Vanmechelen E, Trojanowski JQ, Lee VM, Van Broeckhoven C, van der Zee J, et al. TDP-43 as a possible biomarker for frontotemporal lobar degeneration: a systematic review of existing antibodies. Acta Neuropathol Commun. 2015;3:15.  https://doi.org/10.1186/s40478-015-0195-1.CrossRefPubMedPubMedCentralGoogle Scholar
  25. 25.
    Nelson GM, Guynn JM, Chorley BN. Procedure and key optimization strategies for an automated capillary electrophoretic-based immunoassay method. J Vis Exp. 2017;(127).  https://doi.org/10.3791/55911.
  26. 26.
    Alegria-Schaffer A, Lodge A, Vattem K. Performing and optimizing Western blots with an emphasis on chemiluminescent detection. Methods Enzymol. 2009;463:573–99.  https://doi.org/10.1016/S0076-6879(09)63033-0. CrossRefPubMedGoogle Scholar
  27. 27.
    Rath A, Glibowicka M, Nadeau VG, Chen G, Deber CM. Detergent binding explains anomalous SDS-PAGE migration of membrane proteins. Proc Natl Acad Sci U S A. 2009;106(6):1760–5.  https://doi.org/10.1073/pnas.0813167106. CrossRefPubMedPubMedCentralGoogle Scholar
  28. 28.
    R&D systems a biotechne brand. Human/Mouse/Rat TDP-43/TARDBP Antibody - Monoclonal Mouse IgG2A Clone # 671834 - Catalog Number: MAB7778 - https://resources.rndsystems.com/pdfs/datasheets/mab7778.pdf. Rev. 2/7/2018.
  29. 29.
    Steinacker P, Hendrich C, Sperfeld AD, Jesse S, von Arnim CA, Lehnert S, et al. TDP-43 in cerebrospinal fluid of patients with frontotemporal lobar degeneration and amyotrophic lateral sclerosis. Arch Neurol. 2008;65(11):1481–7.  https://doi.org/10.1001/archneur.65.11.1481.CrossRefPubMedPubMedCentralGoogle Scholar
  30. 30.
    Charette SJ, Lambert H, Nadeau PJ, Landry J. Protein quantification by chemiluminescent Western blotting: elimination of the antibody factor by dilution series and calibration curve. J Immunol Methods. 2010;353(1–2):148–50.  https://doi.org/10.1016/j.jim.2009.12.007.CrossRefPubMedGoogle Scholar
  31. 31.
    Fourier A, Dorey A, Perret-Liaudet A, Quadrio I. Detection of CSF 14-3-3 protein in sporadic Creutzfeldt-Jakob disease patients using a new automated capillary Western assay. Mol Neurobiol. 2017.  https://doi.org/10.1007/s12035-017-0607-2.
  32. 32.
    Mattsson N, Andreasson U, Persson S, Carrillo MC, Collins S, Chalbot S, et al. CSF biomarker variability in the Alzheimer’s Association quality control program. Alzheimers Dement. 2013;9(3):251–61.  https://doi.org/10.1016/j.jalz.2013.01.010.CrossRefPubMedPubMedCentralGoogle Scholar
  33. 33.
    Bennion Callister J, Pickering-Brown SM. Pathogenesis/genetics of frontotemporal dementia and how it relates to ALS. Exp Neurol. 2014;262(Pt B):84–90.  https://doi.org/10.1016/j.expneurol.2014.06.001.CrossRefPubMedPubMedCentralGoogle Scholar

Copyright information

© Springer-Verlag GmbH Germany, part of Springer Nature 2018

Authors and Affiliations

  • Anthony Fourier
    • 1
    • 2
    Email author
  • Jean Escal
    • 1
    • 2
  • Emilien Bernard
    • 3
  • Ingolf Lachman
    • 4
  • Armand Perret-Liaudet
    • 1
    • 2
    • 5
  • Pascal Leblanc
    • 6
  • Isabelle Quadrio
    • 1
    • 2
    • 5
  1. 1.Neurochemistry Laboratory, Biochemistry Department, Centre de Biologie et Pathologie EstHospices Civils de LyonBronFrance
  2. 2.BIORAN Team, Lyon Neuroscience Research Center, CNRS UMR 5292 – INSERM U1028Université de Lyon – Université Claude BernardBronFrance
  3. 3.Hôpital Neurologique Pierre WertheimerBronFrance
  4. 4.AJ Roboscreen GmbHLeipzigGermany
  5. 5.Center for Memory Resources and Research, Hospices Civils de Lyon, Charpennes HospitalLyon 1 UniversityVilleurbanneFrance
  6. 6.Institut NeuroMyoGène (INMG), CNRS UMR5310 - INSERM U1217Université de Lyon - Université Claude BernardLyonFrance

Personalised recommendations