Comparison of 2 Luminex-based Multiplexed Protein Assays for Quantifying Microglia Activation and Inflammatory Proteins

Report No. ARL-TR-7645
Authors: Lee Campbell; Emily Wires; Brandon Harvey; Keith Whitaker
Date/Pages: April 2016; 30 pages
Abstract: The complex injury cascades associated with mild traumatic-brain injury partly involve inflammation in the brain that is triggered by the release of chemokines and cytokines from the brainâ??s resident immune cells, microglia. The technical ability to quantify these small molecules from biological samples is critical toward new technology that limits the damage following a concussive event. This projectâ??s objective was to test the feasibility of using multiplexed protein assays based on Luminex microspheres to fulfil the requirement to quantify inflammatory proteins from biological samples derived from rats. Two vendors, R&D Systems and EMD Millipore, market antibodies conjugated with Luminex fluorescently labeled microspheres that can be used to quantify inflammatory biomarkers directly from samples of plasma, cell cultures, tissue homogenates, cerebral spinal fluid, and other tissues. The single kit from R&D Systems removed many sources of human error and provides a diluent to generate a single standard curve for a variety of sample types. EMD Millipore has many more products available to detect a wide range of analytes in rat samples, but these products are separated into different kits. Without optimization, the products from both vendors were only capable of relative quantification of fluorescence instead of true quantification of concentration.
Distribution: Approved for public release
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Last Update / Reviewed: April 1, 2016