A biomarker panel has been designed based on previous investigations by our group in older populations, including older adults with neurodegenerative conditions [10 (link),38 (link),39 (link),40 (link),41 (link),42 (link),43 (link)]. Markers of systemic inflammation will be assayed as described elsewhere [40 (link)]. Briefly, a set of 27 pro- or anti-inflammatory mediators (e.g., cytokines, chemokines, and growth factors) will be measured in serum samples using the Bio-Plex Pro™ Human Cytokine 27-plex Assay kit (#M500KCAF0Y, Bio-Rad, Hercules, CA, USA) (Table 2). Experiments will be run on a Bio-Plex® System with Luminex xMap Technology (Bio-Rad) and data will be acquired on the Bio-Plex Manager Software 6.1 (Bio-Rad) with instrument default settings. Outliers will be automatically removed by optimization of standard curves across all analytes and results will be obtained as concentration (pg/mL).
Traditional and automated enzyme-linked immunosorbent assays will be run to quantify serum levels of amyloid beta (aa1-42) (#DAB142, R&D Systems, Minneapolis, MN, USA), neurofilament light polypeptide (#SPCKB-PS-002448-000190, R&D Systems), Tau (#NBP2-62749, Novus Biologicals, Littleton, CO, USA), and Tau [p Ser739] (#NBP2-66711, Novus Biologicals) proteins according to the manufacturer’s instructions.
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