Tau and Beyond: Proteomic Profiling of Neurodegenerative Diseases with a 120-Plex CNS Disease Panel

ABSTRACT

The global rise of an increasingly aging population has heightened the need for biomarkers for the early detection of aging-related diseases including neurodegenerative diseases (NDD). Fluid-based biomarkers, especially blood-based biomarkers that can be detected in a minimally invasive manner, are highly valuable. However, progress in translational research has been impeded by a lack of highly multiplexed and ultrasensitive proteomic technologies. Addressing this challenge, we developed NULISA™, which employs DNA-conjugated antibodies to convert immunocomplexes into reporter DNA molecules that can be analyzed with real-time PCR (qPCR) or next-generation sequencing (NGS) to achieve attomolar limit of detection (low to sub-fg/mL) and 100s-plex capability. Here we report the development of a 120-plex NULISA panel – CNS Disease Panel 120 – for comprehensive profiling of NDD. This panel includes well-established biomarkers such as neurofilament light, alpha-synuclein and multiple phosphorylated Tau proteoforms (p-Tau181, p-Tau217 and p-Tau231) as well as other proteins implicated in NDD. We evaluated the performance of this assay with blood (n=38) and cerebrospinal fluid (CSF) (n=29) samples from healthy controls and NDD patients. With just 10 mL plasma or CSF, NULISA demonstrated high sensitivity (~95% detectability in plasma and ~80% in CSF) and high precision (median CV ~6.0%). Linear model analysis identified both known and novel proteins with significant differences between disease and age-matched controls. With full automation in a high throughput system (ARGO HT™), NULISA CNS Disease Panel 120 holds great promise to reveal new biological insights into aging and disease mechanisms and enable biomarker discovery through large cohort and population-based studies.