Multiplexed femtomolar quantitation of XXXX cytokines in a fluoropolymer microcapillary film.

@entity1 @entity32 @entity650 @entity1356 @entity310 @entity281
UNASSIGNED: Sensitive quantitation of multiple cytokines can provide important diagnostic information during @entity281 , @entity32 and immunopathology. In this study sensitive immunoassay detection of @entity1 cytokines @entity1356 , @entity650 , IL-12p70 and @entity310 is shown for singleplex and multiplex formats using a novel miniaturized ELISA platform. The platform uses a disposable plastic multi-syringe aspirator (MSA) integrating 8 disposable fluoropolymer microfluidic test strips, each containing an array of ten 200  m mean i.d. microcapillaries coated with a set of monoclonal antibodies. Each MSA device thus performs 10 tests on 8 samples, delivering 80 measurements. Unprecedented levels of sensitivity were obtained with the novel fluoropolymer microfluidic material and simple colorimetric detection in a flatbed scanner. The limit of detection for singleplex detection ranged from 2.0 to 15.0 pg ml(-1), i.e. 35 and 713 femtomolar for singleplex cytokine detection, and the intra- and inter-assay coefficient of variation (CV) remained within 10%. In addition, a triplex immunoassay was developed for measuring @entity1356 , IL-12p70 and @entity310 simultaneously from a given sample in the pg ml(-1) range. These assays permit high sensitivity measurement with rapid <15 min assay or detection from undiluted blood serum. The portability, speed and low-cost of this system are highly suited to point-of-care testing and field diagnostics applications.
