US HUPO 2024: ARGO™ HT System: An Automated, Ultra-Sensitive and High-Throughput Proteomics Platform for the Blood Proteome

Abstract

Purpose: Existing proteomic platforms often make tradeoffs between sensitivity and multiplexing capabilities and rely on labor-intensive processes. Here we introduce ARGO HT, an automated high-throughput proteomics platform specifically designed for the NUcleic acid Linked Immuno-Sandwich Assay (NULISA™) technology, a novel immunoassay with attomolar sensitivity and high multiplexing capability. We developed a 250-plex inflammation panel and a 120-plex central nervous system (CNS) disease panel and evaluated their performance on the ARGO HT system.

Methods: The 250-plex inflammation panel contained a wide range of cytokines, chemokines, and other immune-related proteins. It was used to profile the blood of patients with autoimmune diseases (n=21) and COVID-19 at multiple time points (n=46). The 120-plex CNS panel included well-established biomarkers like neurofilament light (NfL) and phosphorylated Tau (p-Tau181, p-Tau217 and p-Tau231) as well as other targets of high relevance to neurodegenerative diseases (NDDs). It was used to analyze plasma (n=38) and cerebrospinal fluid (CSF) (n=29) samples from NDD patients and age-matched controls. Linear model analysis was performed to identify differentially abundant targets between disease and control groups. Coefficient of variation (CV) was calculated for the sample control, and target detectability above the limit of detection (LOD) was calculated for each sample set.

Results: Using 10uL samples, the inflammation and CNS disease panels demonstrated a median CV <10%. Both panels detected over 95% of the targets in plasma, and the CNS panel showed an ~80% target detectability in CSF. Linear regression analysis revealed significant differences in the abundance of both known and novel proteins between disease and control groups.

Conclusion: The benchtop ARGO system fully automates the NULISA technology and demonstrates ultrahigh sensitivity and precision. With the integration of reagents, instrumentation and software ARGO HT democratizes high-throughput proteomic analysis, opening the door to the low-abundance proteome for the discovery of novel biomarkers for early disease detection and therapeutic targets.