Generation and characterization of monoclonal antibodies against pathologically phosphorylated TDP-43

PLoS One. 2024 Apr 18;19(4):e0298080. doi: 10.1371/journal.pone.0298080. eCollection 2024.

Abstract

Inclusions containing TAR DNA binding protein 43 (TDP-43) are a pathological hallmark of frontotemporal dementia (FTD) and amyotrophic lateral sclerosis (ALS). One of the disease-specific features of TDP-43 inclusions is the aberrant phosphorylation of TDP-43 at serines 409/410 (pS409/410). Here, we developed rabbit monoclonal antibodies (mAbs) that specifically detect pS409/410-TDP-43 in multiple model systems and FTD/ALS patient samples. Specifically, we identified three mAbs (26H10, 2E9 and 23A1) from spleen B cell clones that exhibit high specificity and sensitivity to pS409/410-TDP-43 peptides in an ELISA assay. Biochemical analyses revealed that pS409/410 of recombinant TDP-43 and of exogenous 25 kDa TDP-43 C-terminal fragments in cultured HEK293T cells are detected by all three mAbs. Moreover, the mAbs detect pS409/410-positive TDP-43 inclusions in the brains of FTD/ALS patients and mouse models of TDP-43 proteinopathy by immunohistochemistry. Our findings indicate that these mAbs are a valuable resource for investigating TDP-43 pathology both in vitro and in vivo.

MeSH terms

  • Amyotrophic Lateral Sclerosis* / genetics
  • Animals
  • Antibodies, Monoclonal
  • DNA-Binding Proteins / genetics
  • Frontotemporal Dementia* / pathology
  • HEK293 Cells
  • Humans
  • Mice
  • TDP-43 Proteinopathies*

Substances

  • Antibodies, Monoclonal
  • DNA-Binding Proteins

Supplementary concepts

  • Frontotemporal Dementia With Motor Neuron Disease