Quantifying α-Synuclein in cerebrospinal fluid could be challenging because of the multiple structural isoforms and other post-translational modifications (phosphorylation) the molecule can undergo.
Alternative splicing and dysregulated protein degradation can render the molecule into a variety of conformations. All of these factors could complicate the development of a robust immunoassay to quantitate α-Synuclein.
The specificity and selectivity of antibodies used must be carefully examined in order to assess what isoforms of the molecule are being analyzed.
Sample diluents and other non-critical reagents that are included in the commercially available kits are not always readily usable for biomarker analysis in clinical programs.
Commercially available kits that are primarily built for research use only (RUO) may need to undergo substantial optimization depending on how the biomarker data (context of use) will be used in the drug development program.
In conclusion, we have optimized the sample diluent, validated our total α-Synuclein SiMoA assay, and also assessed biological variability among normal and Parkinson’s disease patient samples to demonstrate clinical utility of the assay.
Upon completion, participants will be able to carefully examine the clinical utility of the Research Use Only (RUO) kits provided by several vendors for Biomarker studies.
Upon completion, participants will be able to understand how can they tweak the reagents, dilutions and procedure recommended by the kit manufacturer to built a robust assay for clinical utility.
Upon completion, participants will be able to understand how to adjust the sample diluents to enhance the signal to noise ratios, determine the minimum required dilution and minimize interference and cross-reactivity in method development.