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Multiplexed femtomolar quantitation of human cytokines in a fluoropolymer microcapillary film

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journal contribution
posted on 04.08.2015, 11:47 by Nuno Reis, Ana P. Castanheira, Ana I. Barbosa, Alexander D. Edwards
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.

Funding

Ana P. Castanheira was sponsored by the ERASMUS program.

History

School

  • Science

Department

  • Chemistry

Published in

Analyst

Citation

REIS, N.M. ... et al, 2015. Multiplexed femtomolar quantitation of human cytokines in a fluoropolymer microcapillary film. Analyst, 140, pp.5609-5618

Publisher

The Royal Society of Chemistry

Version

VoR (Version of Record)

Publisher statement

This work is made available according to the conditions of the Creative Commons Attribution 3.0 Unported (CC BY 3.0) licence. Full details of this licence are available at: http://creativecommons.org/licenses/by/3.0/

Publication date

2015

Notes

This is an Open Access article licensed under a Creative Commons Attribution 3.0 Unported Licence.

ISSN

1938-3312

Language

en

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