Development of a Sensitive and Reliable Meso Scale Discovery-Based Electrochemiluminescence Immunoassay to Quantify TDP-43 in Human Biofluids

Biosensors (Basel). 2024 Nov 28;14(12):578. doi: 10.3390/bios14120578.

Abstract

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 4-20,000 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.

Keywords: ALS; MSD; TDP-43; biomarker; immunoassay; neurodegenerative disease.

MeSH terms

  • Amyotrophic Lateral Sclerosis / diagnosis
  • Biomarkers / blood
  • Biosensing Techniques
  • DNA-Binding Proteins* / metabolism
  • Electrochemical Techniques
  • Humans
  • Immunoassay
  • Limit of Detection
  • Luminescent Measurements
  • Reproducibility of Results

Substances

  • DNA-Binding Proteins
  • TARDBP protein, human
  • Biomarkers