Electrochemiluminescence platform for the detection of C-reactive proteins: application of recombinant antibody technology to cardiac biomarker detection

O’Reilly, Emmet J.; Conroy, Paul J.; Hearty, Stephen; Keyes, Tia E.; O’Kennedy, Richard; Forster, Robert J.; Dennany, Lynn

doi:10.1039/c5ra08450d

Cited by 35 publications

(27 citation statements)

References 18 publications

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“…Advances in nanostructured materials such as quantum and carbon nano-dots have played a significant role in establishing ECL as a robust and efficient tool for the detection of both chemical and biological species. However, [Ru(bpy)3] 2+ or closely related analogues continue to dominate the field [12,13]. This is primarily due to the high sensitivity, relatively high ECL efficiency and wide dynamic range afforded by transition metal based luminophores [1,4,14,15].…”

Section: Introductionmentioning

confidence: 99%

Deactivation of the ruthenium excited state by enhanced homogeneous charge transport: Implications for electrochemiluminescent thin film sensors

O’Reilly

Keyes

Forster

et al. 2018

Electrochemistry Communications

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The Electrochemiluminescent (ECL) performance of three ruthenium based metallopolymer platforms with different homogeneous charge transfer diffusion coefficients (DCT) is reported. Significantly, simultaneous detection of light and current in tandem with steady state photoluminescence studies demonstrate that increasing the rate of Ru 3+ production via enhanced charge transport results in a decrease in ECL intensity of up to 82% when the concentration of the co-reactant, sodium oxalate, is low, i.e., sub-mM. Spectroelectrochemical studies demonstrate that for maximum sensitivity to be obtained, the electroactive properties of the polymeric support matrix need to be considered in tandem with luminophore, analyte and co-reactant concentrations.

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Section: Introductionmentioning

confidence: 99%

Deactivation of the ruthenium excited state by enhanced homogeneous charge transport: Implications for electrochemiluminescent thin film sensors

O’Reilly

Keyes

Forster

et al. 2018

Electrochemistry Communications

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“…In all cases, the rate-determining steps for the overall process include (i) the kinetics underlying the heterogeneous electron transfer reactions of the coreactant, (ii) the stability of coreactant radicals, and (iii) the distribution of ECL luminophores. These factors critically affect the ECL mechanism and the final signal efficiency 16,17 . Here, we identify a mechanism for boosting ECL emission by optimizing the distribution of radicals and luminophores; moreover, we demonstrate the possibility of enhancing signals for quantifying important markers.…”

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confidence: 99%

Insights into the mechanism of coreactant electrochemiluminescence facilitating enhanced bioanalytical performance

et al. 2020

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Electrochemiluminescence (ECL) is a powerful transduction technique with a leading role in the biosensing field due to its high sensitivity and low background signal. Although the intrinsic analytical strength of ECL depends critically on the overall efficiency of the mechanisms of its generation, studies aimed at enhancing the ECL signal have mostly focused on the investigation of materials, either luminophores or coreactants, while fundamental mechanistic studies are relatively scarce. Here, we discover an unexpected but highly efficient mechanistic path for ECL generation close to the electrode surface (signal enhancement, 128%) using an innovative combination of ECL imaging techniques and electrochemical mapping of radical generation. Our findings, which are also supported by quantum chemical calculations and spin trapping methods, led to the identification of a family of alternative branched amine coreactants, which raises the analytical strength of ECL well beyond that of present state-of-the-art immunoassays, thus creating potential ECL applications in ultrasensitive bioanalysis.

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“…Dependence of ECL signals from the QD composite films on the concentration of 20 µM to 4 mM Hcy in 1 mM K2S2O8 in 0.1 M PBS (black represents blank control) at 100 mV s -1 over the potential range -1.20 ≤ ν ≤ 0.00 V vs. Ag/AgCl. Inset shows the trend of maximum ECL signal at ~-1.00 V vs. Ag/AgCl for this data.ECL response mechanism of homocysteineHcy inhibits the ECL signal through quenching of the radical precursor species, SO4 •following equations 1 and 2 21,22. …”

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confidence: 99%

Cathodic Quantum Dot Facilitated Electrochemiluminescent Detection in Blood

2018

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The expansion of electrochemical sensors to biomedical applications at point of care requires these sensors to undergo analysis without any pre-treatment of extraction. This poses a major challenge for all electrochemical sensors including electrochemiluminescent (ECL) based sensors. ECL offers many advantages for biomedical applications however; obtaining results from complex matrices has proven to be a large hurdle for the application of ECL sensors within this field. This work demonstrates the potential of cathodic ECL to detect and quantify homocysteine with 0.1 nM limit of detection, which is associated with hyperhomocysteinemia, in blood. This near infrared quantum dot (NIR QD) based ECL sensor displays good linearity allowing for rapid detection and providing a basis for exploitation of ECL based sensors for biomedical diagnostics utilising homocysteine as a model cathodic co-reactant. This work will lay the foundations for future developments in biosensing and imaging fields and stands as an initial proof of concept for the utilization of cathodic ECL technologies for biomedical applications once the limits of detection within clinically relevant levels has been achieved. This work illustrates the potential of cathodic ECL sensors, using Hcy as a model complex, for the detection of biomolecules. EXPERIMENTAL Materials Qdot® 800 ITK™ organic quantum dots, (1 μM in decane) were obtained from Invitrogen. Lumidot™ 560 and 640 nm QDs, (5 mg/mL in toluene), chitosan (medium molecular weight, 75-85 % de-acetylated), and all other chemicals were purchased from Sigma-Aldrich. All solutions were prepared in milli-Q water (18 mΩ cm). Bovine whole blood samples utilised within this study were obtained from Wishaw Abattoir Ltd following University of Strathclyde ethical approval. These were stored in aliquots at-20 ºC. Aliquots were defrosted at room temperature on the day of analysis and used immediately. Instrumentation A CH instrument model 760D electrochemical analyser using a standard 3 electrode setup including a 3 mm diameter GC working electrode, Pt wire counter electrode and Ag/AgCl 3 M KCl reference electrode purchased from IJ Cambria Scientific Ltd (UK) was utilised to record all electrochemical measurements. GC electrodes were cleaned following the pro-ASSOCIATED CONTENT Supporting Information Supporting material includes ECL responses for the interactions with reactive oxygen species (Figure S1), ECL responses for 1 mM K2S2O8 in blood with the Stern-Volmer and modified Stern-Volmer plots for this data (Figure S2). The ECL dependence for the interferents given in Figure 6 (Figure S3). Chromatographic experimental details are also included (Figure S4). This material is available free of charge via the Internet at http://pubs.acs.org.

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Electrochemiluminescence platform for the detection of C-reactive proteins: application of recombinant antibody technology to cardiac biomarker detection

Cited by 35 publications

References 18 publications

Deactivation of the ruthenium excited state by enhanced homogeneous charge transport: Implications for electrochemiluminescent thin film sensors

Deactivation of the ruthenium excited state by enhanced homogeneous charge transport: Implications for electrochemiluminescent thin film sensors

Insights into the mechanism of coreactant electrochemiluminescence facilitating enhanced bioanalytical performance

Cathodic Quantum Dot Facilitated Electrochemiluminescent Detection in Blood

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