Epitope mapping of antibodies against TDP-43 and detection of protease-resistant fragments of pathological TDP-43 in amyotrophic lateral sclerosis and frontotemporal lobar degeneration

Biochem Biophys Res Commun. 2012 Jan 6;417(1):116-21. doi: 10.1016/j.bbrc.2011.11.066. Epub 2011 Nov 22.

Abstract

TAR DNA-binding protein of 43 kDa (TDP-43) is the major component of the intracellular inclusions in amyotrophic lateral sclerosis (ALS) and frontotemporal lobar degeneration (FTLD). Here, we show that both monoclonal (60019-2-Ig) and polyclonal (10782-2-AP) anti-TDP-43 antibodies recognize amino acids 203-209 of human TDP-43. The monoclonal antibody labeled human TDP-43 by recognizing Glu204, Asp205 and Arg208, but failed to react with mouse TDP-43. The antibodies stained the abnormally phosphorylated C-terminal fragments of 24-26 kDa in addition to normal TDP-43 in ALS and FTLD brains. Immunoblot analysis after protease treatment demonstrated that the epitope of the antibodies (residues 203-209) constitutes part of the protease-resistant domain of TDP-43 aggregates which determine a common characteristic of the pathological TDP-43 in both ALS and FTLD-TDP. The antibodies and methods used in this study will be useful for the characterization of abnormal TDP-43 in human materials, as well as in vitro and animal models for TDP-43 proteinopathies.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Amino Acid Sequence
  • Amyotrophic Lateral Sclerosis / metabolism*
  • Animals
  • Antibodies / chemistry
  • Antibodies / immunology*
  • Antibodies, Monoclonal / chemistry
  • Antibodies, Monoclonal / immunology
  • Brain / immunology
  • Brain / metabolism
  • Brain Chemistry
  • DNA-Binding Proteins / chemistry
  • DNA-Binding Proteins / immunology*
  • DNA-Binding Proteins / metabolism
  • Epitope Mapping*
  • Frontotemporal Lobar Degeneration / metabolism*
  • Humans
  • Mice
  • Molecular Sequence Data
  • Peptide Hydrolases / chemistry

Substances

  • Antibodies
  • Antibodies, Monoclonal
  • DNA-Binding Proteins
  • Peptide Hydrolases