A Novel Quantitative, Multianalyte Immunoassay to Detect Neuroinflammation following Traumatic Brain Injury

M. Anderson, G. Hickey, I. O’Brien, P. Younge, J. David, L. Leong



Public awareness of Traumatic Brain Injury (TBI) has been increasing due to its appearance in vast array of groups including children, military personnel, and the elderly. This has created a need for the early detection of circulating biomarkers that reflect a traumatized state. Neuronal injury typically stimulates the release of proinflammatory mediators that induce a neuroinflammatory response. It is thought that this response is beneficial as it lays the groundwork for subsequent tissue repair and restoration. A prolonged, unregulated release of proinflammatory cytokines and chemokines, though, has the potential to damage uninvolved neurons and compromise brain function. The ability to distinguish between short-term and prolonged, or long-term, inflammation is thus desirable, particularly when using a marginally-invasive method that utilizes serum or plasma. The study presented here evaluated several neuroinflammatory markers including; Chitinase 3-like 1 (CHI3L1), CXCL8/IL-8, CXCL10/IP-10, ICAM-1/CD54, IL-1 beta/IL-1F2, IL-6, IL-10, and TNF-alpha. The Simple Plex™ (ProteinSimple) assay employed is a novel, quantitative, multianalyte immunoassay platform that delivers high precision and accuracy while using < 25 μL of sample. This platform measures up to four analytes simultaneously from a single, small sample with very high sensitivity. The microfluidic-based system allows parallel single analyte detection and reduces the non-specific antibody interactions often observed in other traditional multiplex platforms. The assay is also a closed system and the whole process is automated within a single cartridge, thus removing potential user variability. Furthermore, results are generated in just over one hour. Our studies suggest this platform may represent a highly efficient method for the detection of pro- and anti-inflammatory cytokines following TBI.