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Ongoing Development and Characterization of Tools and Resources for Parkinson’s Disease Research

In recent years, The Michael J. Fox Foundation for Parkinson's Research (MJFF) has endeavored to accelerate progress in Parkinson's disease (PD) research and therapeutic development by decreasing barriers and increasing access to critical resources including verified DNA plasmids, quality antibodies, robust assays and viral vectors encoding target genes of interest. As these have been generated and distributed, ongoing characterization efforts are revealing the utility of each of these tools in a number of routine laboratory applications.

MJFF has begun generating viral vectors encoding several PD targets of interest (including alpha-synuclein and LRRK2).   Thus far, AAV2-alpha-synuclein and AAV5-alpha-synuclein vectors have yielded alpha-synuclein expression levels in vivo at early time points in keeping with published results.  These tools will help facilitate studies in cellular systems and animal models that will enable the research community to better understand a biological target's relevance to PD.

To date, MJFF has generated more than a dozen antibodies against PD-associated targets, many of which are now widely available to the research community.  MJFF has also led a characterization study of existing LRRK2 antibodies and revealed those which are optimal in specific applications including immunohistochemistry, immunoblot and immunoprecipitation applications.  MJFF is generating and characterizing additional antibodies, including phosphorylated forms of LRRK2, oxidized DJ-1, PINK1 and phosphorylated or aggregated forms of alpha-synuclein.

Finally, MJFF is also developing ELISA methods to quantify levels of LRRK2 protein in biological samples.  Utilizing a truncated form of LRRK2 as a protein standard in a sandwich ELISA, eight antibodies were evaluated as coat or detection protein pairs.  Ongoing optimization of conditions in a number of matrices is revealing that antibodies at Covance and Neuromab have superior performance.  This assay will enable investigators to further characterize tissue and other biological samples for changes in LRRK2 protein levels in human tissue and to correlate these changes with disease progression.

MJFF is confident that the availability of these reagents to the entire research community will facilitate understanding of PD and accelerate therapeutic development. Further, MJFF can replicate this strategy for generating additional reagents where decreased barriers to access and limited resources hinder progress in PD research efforts.

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