Multiplexed femtomolar quantitation of human cytokines in a fluoropolymer microcapillary film

Castanheira, A. P., Barbosa, A. I., Edwards, A. D. ORCID: https://orcid.org/0000-0003-2369-989X and Reis, N. M. (2015) Multiplexed femtomolar quantitation of human cytokines in a fluoropolymer microcapillary film. Analyst, 140 (16). pp. 5609-5618. ISSN 0003-2654 doi: 10.1039/C5AN00238A

Abstract/Summary

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 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.

Item Type	Article
URI	https://reading-clone.eprints-hosting.org/id/eprint/41185
Identification Number/DOI	10.1039/C5AN00238A
Refereed	Yes
Divisions	Life Sciences > School of Chemistry, Food and Pharmacy > School of Pharmacy > Pharmaceutics Research Group
Publisher	Royal Society of Chemistry
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