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Title: Multiplexed femtomolar quantitation of human cytokines in a fluoropolymer microcapillary film
Authors: Reis, Nuno M.
Castanheira, Ana P.
Barbosa, Ana I.
Edwards, Alexander D.
Issue Date: 2015
Publisher: The Royal Society of Chemistry
Citation: REIS, N.M. ... et al, 2015. Multiplexed femtomolar quantitation of human cytokines in a fluoropolymer microcapillary film. Analyst, 140, pp.5609-5618
Abstract: Sensitive quantitation of multiple cytokines can provide important diagnostic information during infection, inflammation and immunopathology. In this study sensitive immunoassay detection of human cytokines IL-1β, IL-6, IL-12p70 and TNFα 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 μm 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, 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 IL-1β, IL-12p70 and TNFα simultaneously from a given sample in the pg/ml 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.
Description: This is an Open Access article licensed under a Creative Commons Attribution 3.0 Unported Licence.
Sponsor: Ana P. Castanheira was sponsored by the ERASMUS program.
Version: Published
DOI: 10.1039/C5AN00238A
URI: https://dspace.lboro.ac.uk/2134/18401
Publisher Link: http://dx.doi.org/10.1039/C5AN00238A
ISSN: 1938-3312
Appears in Collections:Published Articles (Chemistry)

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