Quantitative Measurements of LRRK2 in Human Cerebrospinal Fluid Demonstrates Increased Levels in G2019S Patients

Frontiers in Neuroscience
Omar S MabroukDanielle L Graham

Abstract

Leucine-rich repeat kinase 2 (LRRK2) mutations are among the most significant genetic risk factors for developing late onset Parkinson's disease (PD). To understand whether a therapeutic can modulate LRRK2 levels as a potential disease modifying strategy, it is important to have methods in place to measure the protein with high sensitivity and specificity. To date, LRRK2 measurements in cerebrospinal fluid (CSF) have used extracellular vesicle enrichment via differential ultracentrifugation and western blot detection. Our goal was to develop a methodology which could be deployed in a clinical trial, therefore throughput, robustness and sensitivity were critical. To this end, we developed a Stable Isotope Standard Capture by Anti-peptide Antibody (SISCAPA) assay which is capable of detecting LRRK2 from 1 ml of human CSF. The assay uses a commercially available LRRK2 monoclonal antibody (N241A/34) and does not require extracellular vesicle enrichment steps. The assay includes stable isotope peptide addition which allows for absolute quantitation of LRRK2 protein. We determined that the assay performed adequately for CSF measurements and that blood contamination from traumatic lumbar puncture does not pose a serious analytical cha...Continue Reading

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Citations

Oct 6, 2020·Frontiers in Neuroscience·Hardy J RideoutJean-Marc Taymans
Sep 10, 2020·Frontiers in Neuroscience·Kaela Kelly, Andrew B West
Oct 18, 2020·Translational Neurodegeneration·Christian U von LinstowPatrik Brundin
Jul 9, 2021·Annual Review of Neuroscience·Gabriel E Vázquez-Vélez, Huda Y Zoghbi

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Methods Mentioned

BETA
antisense oligonucleotide
enzyme-linked immunosorbent assay
ELISA
immunoprecipitation
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Software Mentioned

MJFF
SISCAPA
Proteome Discoverer
SICAPA
ChromaLink Molar Ratio MSR )
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