Objective/Rationale:
Mutations in the DJ-1 gene are associated with an early-onset familial form of PD and DJ-1 protein has been shown to be lower in the CSF of sporadic PD compared to controls. DJ-1 protects nerve cells against oxidative stress. Oxidative stress has been implicated in the pathogenesis of PD. Oxidized DJ-1 may be a more sensitive biomarker for oxidative stress associated with the development and progression of PD. We have developed an antibody against an oxidized form of DJ-1 by immunizing pre-clinical models with oxidized peptide containing C106 region of DJ-1. The goals of this study are 1) to affinity purify the polyclonal antibodies to increase selectivity and sensitivity of the antibodies and 2) to generate monoclonal antibodies from the same pre-clinical mdoels as a more stable reagent for future development of biomarker assays.
Project Description:
Polyclonal antibodies from a few different rabbits will be affinity purified by both positive selection against oxidized form of DJ-1 peptide and negative selection against non-oxidized DJ-1. In addition, Monoclonal antibodies will generated from the spleen of the pre-clinical models by generating hybridomas and these antibodies will be screened for specificity and avidity.
Relevance to Diagnosis/Treatment of Parkinson’s Disease:
The identification of these biomarkers for PD has the potential to monitor PD progression, which will improve patient care by providing a metric for treatment efficacy. In addition, PD biomarkers facilitate the drug discovery process and aid in the recruitment and performance of clinical trials.
Anticipated Outcome:
Successful completion of the goals of this project will result in the development of new monoclonal and polyclonal antibodies against oxidized DJ-1. As DJ-1 may play an important role in PD pathogenesis, new tools for detecting oxidized DJ-1 may also contribute to an improved understanding of PD.