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We report a novel method to analyze multiple cytokines/chemokines in 5 μL of BALF in mice. The methodology relies on micro-volume wall-less immunoassays combined with Luminex magnetic bead multiplex technology. This new method has been demonstrated with good validity, veracity, and sensitivity by comparing it with the existing conventional Luminex method, as well comparing the values of BALF cytokines/chemokines from functional inflammatory stimulated mice with that of healthy mice. Compared with conventional Luminex method, it also showed obvious advantages in some other aspects such as the sample volume was minimized to 5 μL, the consumption of reagents and beads was reduced by a factor of 5 and the total cost of experiments was significantly reduced by 80%, which would have wide applications in the field of multiple cytokines/chemokines analysis. [ABSTRACT FROM AUTHOR]

Rapid and high-throughput analytical techniques for IgE that requires a small serum amount are very important, especially for pediatric patients. In these patients, blood is collected from veins, which is painful compared to fingertip blood collection. Herein, a novel microfluidic system capable of high-throughput parallel analyses of allergen-specific IgE from small amounts of patient serum was successfully developed. A six-plex immunoassay was constructed within a microfluidic chip, and the entire system was validated using samples from clinical patients. Major antigens from house dust mite ( Dermatophagoides farinae and Blomia tropicalis ), cat ( Felis domesticus ), fungus ( Cladosporium herbarum ), ragweed ( Humulus japonicas ), and tree pollen ( Platanus acerifolia ) were used as analysis targets. Sample consumption decreased to <0.05 µL compared with the 480 µL serum consumption by fluoroenzyme immunoassay (UniCAP system Pharmacia Diagnostics AB, Uppsala, Sweden), the 50 µL serum consumption by enzyme-linked immune sorbent assay (ELISA), or the 1.5 µL serum consumption by conventional protein chip analysis. Analysis duration, reagent cost, and total cost for each measurement were also considerably decreased. The assay showed good accuracy and sensitivity toward the clinical samples. A significant correlation of allergen-specific IgE levels was found among the microfluidic assay, UniCAP system, and ELISA. [ABSTRACT FROM AUTHOR]