Development of an ultra-sensitive multiplex cytokine assay utilizing a novel digital multi-omics detection technology

Abstract

Abstract Demand for rapid, highly sensitive measurement of proteins and nucleic acids has increased dramatically over the past several years to support biomarker research, drug development and diagnostics. In addition, the ability to interrogate both protein and nucleic acids (multi-omics) in a single sample allows critical information to be obtained relating the transcriptome to expressed protein levels. Despite this, current platforms for multiplex measurement of circulating biomarkers can lack sensitivity and specificity required for analytical and clinical validation of low abundance proteins and nucleic acids. We report the development of an automated microfluidic-based technology capable of multiplexed measurement of protein and nucleic acid biomarkers utilizing a novel approach combining digital and analog detection modalities. Significant improvements in sensitivity and specificity are achieved through use of a bead-based microfluidic array resulting in a spatially-resolved series of single-plex signal amplification reactions. Critically, the isolation of reactions allows multiplex panels to be constructed in a modular manner while eliminating interference that often limits specificity of traditional multiplex biomarker detection technologies. A specific application of this technology we report here is a multiplexed assay measuring low abundance cytokine biomarkers from serum and plasma samples with utility in a broad range of research and clinical studies.