Quantifying IL-27 and CRP Biomarkers

Blood samples were collected into tubes with EDTA and centrifuged at 2000× g for 15 min at 4 °C. Plasma was frozen and stored at −80 °C until use. For the measurement of IL-27 levels, the enzyme-linked immunosorbent assay (ELISA) (LEGEND MAX™ Human IL-27 ELISA Kit; BioLegend, San Diego, CA, USA) was used. As previously reported [21 (link)], we assessed IL-27 levels at Ochanomizu University according to the manufacturer’s instructions. According to data from the manufacturer, the lowest detection limit of this kit was 0.01 ng/mL. The intra- and interassay coefficients of variation were found to be <6.0% and <5.5%. To measure high-sensitivity C-reactive protein (CRP) levels, a BNII nephelometer (Siemens Healthineers, Tokyo, Japan) was used.