Multiplexed femtomolar quantitation of human cytokines in a fluoropolymer microcapillary film

Ana P. Castanheira, Ana I. Barbosa, Alexander D. Edwards, Nuno M. Reis

Research output: Contribution to journalArticle

15 Citations (Scopus)

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−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 IL-1β, IL-12p70 and TNFα 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.
Original languageEnglish
Pages (from-to)5609-5618
JournalAnalyst
Volume140
Issue number16
DOIs
Publication statusPublished - 21 Aug 2015

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Fluorine containing polymers
Assays
Microfluidics
Syringes
Cytokines
Immunoassay
assay
immunoassay
Plastics
Monoclonal antibodies
Limit of Detection
Interleukin-6
Enzyme-Linked Immunosorbent Assay
Monoclonal Antibodies
Inflammation
Costs and Cost Analysis
Equipment and Supplies
Blood
Infection
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Cite this

Multiplexed femtomolar quantitation of human cytokines in a fluoropolymer microcapillary film. / Castanheira, Ana P.; Barbosa, Ana I.; Edwards, Alexander D.; Reis, Nuno M.

In: Analyst, Vol. 140, No. 16, 21.08.2015, p. 5609-5618.

Research output: Contribution to journalArticle

Castanheira, Ana P. ; Barbosa, Ana I. ; Edwards, Alexander D. ; Reis, Nuno M. / Multiplexed femtomolar quantitation of human cytokines in a fluoropolymer microcapillary film. In: Analyst. 2015 ; Vol. 140, No. 16. pp. 5609-5618.
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