Burgess Hill, United Kingdom
Burgess Hill, United Kingdom

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Pivetal J.,University of Reading | Pereira F.M.,Capillary Film Technology Ltd | Barbosa A.I.,Capillary Film Technology Ltd | Barbosa A.I.,Loughborough University | And 6 more authors.
Analyst | Year: 2017

This study reports for the first time the sensitive colorimetric and fluorescence detection of clinically relevant protein biomarkers by sandwich immunoassays using the covalent immobilisation of antibodies onto the fluoropolymer surface inside Teflon®-FEP microfluidic devices. Teflon®-FEP has outstanding optical transparency ideal for high-sensitivity colorimetric and fluorescence bioassays, however this thermoplastic is regarded as chemically inert and very hydrophobic. Covalent immobilisation can offer benefits over passive adsorption to plastic surfaces by allowing better control over antibody density, orientation and analyte binding capacity, and so we tested a range of different and novel covalent immobilisation strategies. We first functionalised the inner surface of a 10-bore, 200 μm internal diameter FEP microcapillary film with high-molecular weight polyvinyl alcohol (PVOH) without changing the outstanding optical transparency of the device delivered by the matched refractive index of FEP and water. Glutaraldehyde immobilisation was compared with the use of photoactivated linkers and NHS-ester crosslinkers for covalently immobilising capture antibodies onto PVOH. Three clinically relevant sandwich ELISAs were tested against the cytokine IL-1β, the myocardial infarct marker cardiac troponin I (cTnI), and the chronic heart failure marker brain natriuretic peptide (BNP). Overall, glutaraldehyde immobilisation was effective for BNP assays, but yielded unacceptable background for IL-1β and cTnI assays caused by direct binding of the biotinylated detection antibody to the modified PVOH surface. We found NHS-ester groups reacted with APTES-treated PVOH coated fluoropolymers. This facilitated a novel method for capture antibody immobilisation onto fluoropolymer devices using a bifunctional NHS-maleimide crosslinker. The density of covalently immobilised capture antibodies achieved using PVOH/APTES/NHS/maleimide approached levels seen with passive adsorption, and sensitive and quantitative assay performance was achieved using this method. Overall, the PVOH coating provided an excellent surface for controlled covalent antibody immobilisation onto Teflon®-FEP for performing high-sensitivity immunoassays. This journal is © The Royal Society of Chemistry.


Reis N.,Capillary Film Technology Ltd | Reis N.,Loughborough University | Pereira F.,Capillary Film Technology Ltd | Barbosa A.,Capillary Film Technology Ltd | And 5 more authors.
20th International Conference on Miniaturized Systems for Chemistry and Life Sciences, MicroTAS 2016 | Year: 2016

Our research team recently pioneered the development of ultra-sensitive sandwich ELISA in a novel melt-extruded microfluidic material called Microcapillary Film (MCF) [1-4]. Quantitative biomarker assays can be performed on whole blood in fluoropolymer MCF strips without the need of sample preparation [2], with simple optical detection with low-cost optoelectronic components including smartphones [3] that deliver limit-of-detection comparable to state-of-the-art clinical equipment [4]. To date, these assays were performed manually, and no automation of assays in MCF strips has yet been reported. Here, we report for the first time prototypes of two fully automated devices for performing ELISA in fluoropolymer MCF, using integrated low cost optoelectronic components for affordable signal quantitation.


Barbosa A.I.,Loughborough University | Gehlot P.,Loughborough University | Sidapra K.,Loughborough University | Edwards A.D.,Capillary Film Technology Ltd | And 2 more authors.
Biosensors and Bioelectronics | Year: 2015

We present a new, power-free and flexible detection system named MCFphone for portable colorimetric and fluorescence quantitative sandwich immunoassay detection of prostate specific antigen (PSA). The MCFphone is composed by a smartphone integrated with a magnifying lens, a simple light source and a miniaturised immunoassay platform, the Microcapillary Film (MCF). The excellent transparency and flat geometry of fluoropolymer MCF allowed quantitation of PSA in the range 0.9 to 60. ng/ml with<7% precision in 13. min using enzymatic amplification and a chromogenic substrate. The lower limit of detection was further improved from 0.4 to 0.08. ng/ml in whole blood samples with the use of a fluorescence substrate. The MCFphone has shown capable of performing rapid (13 to 22. min total assay time) colorimetric quantitative and highly sensitive fluorescence tests with good %Recovery, which represents a major step in the integration of a new generation of inexpensive and portable microfluidic devices with commercial immunoassay reagents and off-the-shelf smartphone technology. © 2015 The Authors.


Ferreira A.I.,Loughborough University | Castanheira A.P.,Capillary Film Technology Ltd | Chahin R.G.,University of Cambridge | Mackley M.R.,University of Cambridge | And 2 more authors.
17th International Conference on Miniaturized Systems for Chemistry and Life Sciences, MicroTAS 2013 | Year: 2013

This paper reports the miniaturisation of sandwich enzyme-linked immunosorbent assays (ELISA) using a meltextruded fluoropolymer MicroCapillary Film (MCF). A prototype 8-channel*10-microcapillaries device was used for quantitative detection a range of human biomarkers in 30 mm long fluoropolymer MCF strips. This represents a novel combination of ultra-low cost plastic microfluidic strips, low-cost optical detection, short incubation times, pre-loaded reagents and sequential fluid aspiration based on disposable plastic syringes. The observed limit of detection (LOD) was ∼lng/ml for PSA, and 10-60pg/ml for IL-lbeta, IL-6 and IL-12 inflammatory cytokines, typically with CVs ≤ 10% which is comparable with performance of optimised colorimetric microwell-based ELISA.


Barbosa A.I.,Loughborough University | Castanheira A.P.,Capillary Film Technology Ltd | Edwards A.D.,Capillary Film Technology Ltd | Reis N.M.,Loughborough University | Reis N.M.,Capillary Film Technology Ltd
Lab on a Chip - Miniaturisation for Chemistry and Biology | Year: 2014

We present a new concept for rapid and fully portable prostate specific antigen (PSA) measurements, termed "lab-in-a-briefcase", which integrates an affordable microfluidic ELISA platform utilising a melt-extruded fluoropolymer microcapillary film (MCF) containing an array of 10 200 μm internal diameter capillaries, a disposable multi-syringe aspirator (MSA), a sample tray pre-loaded with all of the required immunoassay reagents, and a portable film scanner for colorimetric signal digital quantification. Each MSA can perform 10 replicate microfluidic immunoassays on 8 samples, allowing 80 measurements to be made in less than 15 minutes based on semi-automated operation, without the need of additional fluid handling equipment. The assay was optimised for the measurement of a clinically relevant range of PSA of 0.9 to 60.0 ng ml-1 in 15 minutes with CVs on the order of 5% based on intra-assay variability when read using a consumer flatbed film scanner. The PSA assay performance in the MSA remained robust in undiluted or 12 diluted human serum or whole blood, and the matrix effect could simply be overcome by extending sample incubation times. The PSA "lab-in-a-briefcase" is particularly suited to a low-resource health setting, where diagnostic labs and automated immunoassay systems are not accessible, by allowing PSA measurement outside the laboratory using affordable equipment. © 2014 the Partner Organisations.


Castanheira A.P.,Capillary Film Technology Ltd. | Barbosa A.I.,Loughborough University | Edwards A.D.,Capillary Film Technology Ltd. | Edwards A.D.,University of Reading | And 2 more authors.
Analyst | Year: 2015

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. © The Royal Society of Chemistry 2015.

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