Preparation and application of monoclonal antibodies against oxidized DJ-1. Significant elevation of oxidized DJ-1 in erythrocytes of early-stage Parkinson disease patients

Neurosci Lett. 2009 Nov 6;465(1):1-5. doi: 10.1016/j.neulet.2009.08.074. Epub 2009 Sep 3.

Abstract

DJ-1 was initially identified as a novel oncogene and has recently been found to be a causative gene for a familial form of Parkinson's disease (PD), viz, PARK7. Cysteine residue at position 106 (Cys-106) in DJ-1 was found to be oxidized preferentially under oxidative stress. In the present study, we developed specific antibodies against Cys-106-oxidized DJ-1 using baculovirus particles displaying the surface glycoprotein gp64-fusion protein as the immunizing agent. Western blot analysis combined with two-dimensional gel electrophoresis revealed that these antibodies specifically recognized oxidized DJ-1. Furthermore, we developed a competitive enzyme-linked immunosorbent assay (ELISA) for detecting oxidized DJ-1 and measured blood levels of oxidized DJ-1 in PD patients (n=15). It was observed that the levels of oxidized DJ-1 in erythrocytes of unmedicated PD patients were markedly higher without overlap than those of medicated PD patients and healthy subjects. No significant difference was observed in DJ-1 levels between mediated and unmediated PD patient. These results suggest the oxidative modification of DJ-1 in PD patients and the potential application of the antibody for diagnosis of PD at early-stage.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Aged
  • Analysis of Variance
  • Antibodies / metabolism*
  • Antiparkinson Agents / pharmacology
  • Baculoviridae
  • Blotting, Western
  • Cell Line, Tumor
  • Electrophoresis, Gel, Two-Dimensional
  • Enzyme-Linked Immunosorbent Assay
  • Erythrocytes / metabolism*
  • Female
  • Humans
  • Hydrogen Peroxide / metabolism
  • Intracellular Signaling Peptides and Proteins / blood
  • Intracellular Signaling Peptides and Proteins / immunology*
  • Intracellular Signaling Peptides and Proteins / metabolism*
  • Male
  • Middle Aged
  • Oncogene Proteins / blood
  • Oncogene Proteins / immunology*
  • Oncogene Proteins / metabolism*
  • Oxidation-Reduction
  • Parkinson Disease / blood
  • Parkinson Disease / drug therapy
  • Parkinson Disease / metabolism*
  • Protein Deglycase DJ-1
  • Time Factors

Substances

  • Antibodies
  • Antiparkinson Agents
  • Intracellular Signaling Peptides and Proteins
  • Oncogene Proteins
  • Hydrogen Peroxide
  • PARK7 protein, human
  • Protein Deglycase DJ-1