Objectives: The pursuit of blood biomarkers for neurodegenerative diseases (NDDs) has been hampered by the lack of a proteomic tool with the required sensitivity to detect exceedingly low levels of brain-derived proteins in blood. We recently developed a novel proteomic technology called NULISA™ with attomolar sensitivity and high multiplexing in a fully automated system. In this study, we developed the NULISAseq™ CNS Disease Panel 120, a highly multiplexed assay designed to characterize key hallmarks of NDDs in blood and cerebrospinal fluid (CSF). We evaluated the performance of this panel in plasma and CSF samples from NDD patients and healthy controls.

Methods: A 120-plex panel including known disease markers such as neurofilament light, synuclein A and phosphorylated Tau (p-Tau181, p-Tau217, and p-Tau231) was developed and used to analyze plasma (n=40) and CSF (n=16) samples from NDD patients and age-matched controls. Target detectability and assay precision were assessed, and linear regression analysis was performed for each target for differential abundance between disease and controls.

Results: We developed a CNS disease panel of 120 targets implicated in various pathways and processes characteristic of NDDs. Using 10 µL plasma or CSF, NULISA demonstrated high sensitivity detecting ~94% of the targets in plasma and ~81% in CSF and high precision with median CV of plasma 5.4% and median CV of CSF 9.1%. Linear regression analysis identified both known and novel proteins with significant differences in abundance between disease and age-matched controls.

Conclusion: We developed a comprehensive CNS disease-targeted panel for the NULISA platform to address the growing demand for blood-based biomarker discovery and validation. This study demonstrated the potential of the NULISA CNS disease panel to advance research in blood-based biomarkers for early neurodegenerative disease detection and monitoring and eventually therapeutic intervention.