Novel Flow-Based Multiplex Assay Panel for Quantifying Human Neuroinflammation Biomarkers

Abstract

Abstract The inflammation of neuronal tissues is triggered by various stimuli such as traumatic brain injury, stroke, protein aggregation and pathogens. Neuroinflammation cues such as brain injury lead to protein release and microglia activation, which in turn activate glial cells. Activated glial cells produce both pro- and anti-inflammatory factors to regulate cell migration, growth, tissue repair or cell death. Neuroinflammation is involved in acute brain injury, stroke, and many neurodegenerative diseases, such as Alzheimers’s and Parkinson’s diseases, as well as disruption and permeability of the blood-brain barrier. Measurement of the cytokines, chemokines, growth factors and other neuronal disease biomarkers is essential in understanding the mechanisms of neuronal disease progression, but this is limited by the small sample size of CSF. It is therefore desirable to be able to measure a panel of important neuroinflammation biomarkers simultaneously. We have developed an assay panel targeting neuroinflammation biomarkers using fluorescence-encoded beads that are suitable for use on common lab flow cytometers. It allows for simultaneous quantification of the 13 key targets involved in neuroinflammation including VILIP-1, BDNF, free active TGF-β1, VEGF, IL-6, sTREM-1, β-NGF, IL-18, TNF-α, soluble RAGE, and CX3CL1 (Fractialkine). This panel has been developed with superior assay performance and specificity and has been validated by detecting expected values in biological samples including CSF samples from healthy and disease individuals. This high quality, low cost and easy to use panel will be a useful tool to the neuroinflammation research community.