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

Yoshiro Saito, Takao Hamakubo, Yasukazu Yoshida, Yoko Ogawa, Yasuo Hara, Harutoshi Fujimura, Yasuharu Imai, Hiroko Iwanari, Yasuhiro Mochizuki, Mototada Shichiri, Keiko Nishio, Tomoya Kinumi, Noriko Noguchi, Tatsuhiko Kodama, Etsuo Niki

Research output: Contribution to journalArticlepeer-review

57 Citations (Scopus)

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.

Original languageEnglish
Pages (from-to)1-5
Number of pages5
JournalNeuroscience Letters
Volume465
Issue number1
DOIs
Publication statusPublished - 2009 Nov 6
Externally publishedYes

Keywords

  • Biomarker
  • DJ-1
  • Enzyme-linked immunosorbent assay
  • Erythrocytes
  • Oxidative stress
  • Parkinson's disease

ASJC Scopus subject areas

  • Neuroscience(all)

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