Tu166 - Comparative Analysis of Multiplexed Immunoassay Platforms for Inflammatory Markers in a COVID-19 Cohort

Joanne Beer – Alamar Biosciences, Inc.; Tran Nguyen – Stanford University; Ishara Ariyapala – Alamar Biosciences, Inc.; Tyson Holmes – Stanford University; Wei Feng – Alamar Biosciences, Inc.; Bingqing Zhang – Alamar Biosciences, Inc.; Dwight Kuo – Alamar Biosciences, Inc.; Yuling Luo – Alamar Biosciences, Inc.; Xiao-Jun Ma – Alamar Biosciences, Inc.; Holden Maecker – Stanford University

Abstract Text: In this study, we evaluated three distinct immunoassay platforms designed for the simultaneous detection of multiple inflammatory markers. Our assessment focused on three performance metrics - detectability, correlation, and differential expression - using serum samples from the NIH IMPACC study. The scrutinized platforms included the fluorescent bead-based Luminex assay, the proximity extension-based Olink assay, and the novel proximity ligation assay platform, Alamar NULISAseq. Our findings provide valuable insights, revealing that the Alamar platform exhibited the highest overall detectability, followed by Olink and Luminex. Notably, protein measurement correlations between Alamar and Olink were generally stronger than those observed between either of these platforms and Luminex. Furthermore, we conducted a freeze-thaw experiment using the Alamar assay, demonstrating minimal differences between 1 and 2 freeze-thaw cycles for serum samples. This additional experiment adds a layer of robustness to our findings. Through our study, we offer insights into the comparative performance of these assays, contributing to a refined understanding of their strengths and limitations. This is particularly pertinent when applied to complex biological samples, as exemplified by the sera from the COVID-19 cohort. Our research contributes to the selection of protein assays and facilitates improved data interpretation for future studies involving intricate disease cohorts.