Electrochemical detection of protein glycosylation using lectin and protein–gold affinity interactions

Yadav, Sharda; Carrascosa, Laura G.; Sina, Abu Ali Ibn; Shiddiky, Muhammad J. A.; Hill, Michelle M.; Trau, Matt

doi:10.1039/c6an00528d

Cited by 16 publications

(18 citation statements)

References 28 publications

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“…, in this case the DPV current) generated by the [Fe(CN) 6 ] 3−/4− system present in the electrolyte solution. Stepwise addition of the assay components on the sensor surface, blocks the [Fe(CN) 6 ] 3−/4− redox system from accessing the electrode surface, which in turn decreased the DPV current signal 13, 21, 22, 25 . For example, compared to the bare electrode (Fig.…”

Section: Resultsmentioning

confidence: 99%

“…The electrodes were then incubated with the protein of interest, FAM134B. Differential pulse voltammetry (DPV), in presence of the [Fe(CN) 6 ] 4−/3− redox system, was used to monitor the Faradaic currents generated in each step of this sensing layers 20–22 . The addition of subsequent antibody and protein layers on the electrode surface blocks the [Fe(CN) 6 ] 4−/3− redox system from accessing the surface quite effectively, which results in a decrease in DPV current response.…”

Section: Introductionmentioning

confidence: 99%

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An electrochemical method for sensitive and rapid detection of FAM134B protein in colon cancer samples

Islam

Haque

Yadav

et al. 2017

Sci Rep

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Despite the excellent diagnostic applications of the current conventional immunoassay methods such as ELISA, immunostaining and Western blot for FAM134B detection, they are laborious, expensive and required a long turnaround time. Here, we report an electrochemical approach for rapid, sensitive, and specific detection of FAM134B protein in biological (colon cancer cell extracts) and clinical (serum) samples. The approach utilises a differential pulse voltammetry (DPV) in the presence of the [Fe(CN)6]3−/4− redox system to quantify the FAM134B protein in a two-step strategy that involves (i) initial attachment of FAM134B antibody on the surface of extravidin-modified screen-printed carbon electrode, and (ii) subsequent detection of FAM134B protein present in the biological/clinical samples. The assay system was able to detect FAM134B protein at a concentration down to 10 pg μL−1 in phosphate buffered saline (pH 7.4) with a good inter-assay reproducibility (% RSD = <8.64, n = 3). We found excellent sensitivity and specificity for the analysis of FAM134B protein in a panel of colon cancer cell lines and serum samples. Finally, the assay was further validated with ELISA method. We believe that our assay could potentially lead a low-cost alternative to conventional immunological assays for target antigens analysis in point-of-care applications.

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Section: Resultsmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

An electrochemical method for sensitive and rapid detection of FAM134B protein in colon cancer samples

Islam

Haque

Yadav

et al. 2017

Sci Rep

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“…NPFe 2 O 3 NC with α-human IgG with nanocube to form IgG/Au-NPFe 2 O 3 NC nanocatalysts 50,51. These nanocatalysts were then added to the electrode surface to form immunocomplex with the target p53 autoantibody.…”

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confidence: 99%

Gold-Loaded Nanoporous Ferric Oxide Nanocubes with Peroxidase-Mimicking Activity for Electrocatalytic and Colorimetric Detection of Autoantibody

Yadav²,

et al. 2017

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The enzyme-mimicking activity of iron oxide based nanostructures has provided a significant advantage in developing advanced molecular sensors for biomedical and environmental applications. Herein, we introduce the horseradish peroxidase (HRP)-like activity of gold-loaded nanoporous ferric oxide nanocubes (Au-NPFeONC) for the development of a molecular sensor with enhanced electrocatalytic and colorimetric (naked eye) detection of autoantibodies. The results showed that Au-NPFeONC exhibits enhanced peroxidase-like activity toward the catalytic oxidation of 3,3',5,5'-tertamethylbenzidine (TMB) in the presence of HO at room temperature (25 °C) and follows the typical Michaelis-Menten kinetics. The autoantibody sensor based on this intrinsic property of Au-NPFeONC resulted in excellent detection sensitivity [limit of detection (LOD) = 0.08 U/mL] and reproducibility [percent relative standard deviation (% RSD) = <5% for n = 3] for analyzing p53-specific autoantibodies using electrochemical and colorimetric (naked eye) readouts. The clinical applicability of the sensor has been tested in detecting p53-specific autoantibody in plasma obtained from patients with epithelial ovarian cancer high-grade serous subtype (EOCHGS, number of samples = 2) and controls (benign, number of samples = 2). As Au-NPFeONC possess high peroxidase-like activity for the oxidation of TMB in the presence of HO [TMB is a common chromogenic substrate for HRP in enzyme-linked immunosorbent assays (ELISAs)], we envisage that our assay could find a wide range of application in developing ELISA-based sensing approaches in the fields of medicine (i.e., detection of other biomarkers the same as p53 autoantibody), biotechnology, and environmental sciences.

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“…The first layer formed the profile of the biorecognition platform after adding a redox probe, [37,38]. As the concentration of biomolecule moieties increased, electrons access to the surface were partially hindered, leading to a drop in the current response.…”

Section: Resultsmentioning

confidence: 99%

Cancer biomarker profiling using nanozyme containing iron oxide loaded with gold particles

Akpe

Shiddiky

Brown

et al. 2020

J. R. Soc. Interface.

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Nanozymes are nanomaterials with intrinsic magnetism and superparamagnetic properties. In the presence of an external magnet, nanozyme particles aggregate and redisperse without a foreign attraction. We evaluated the performances of nanozyme by changing the biosensing platforms and substituting other biological variants for a complete cancer assay detection. We investigated the expression of morphological variants in the transmission of signals using an electrochemical method. The signal responses, including signal enhancement with the nanozyme (Au-Fe 2 O 3 ), showed a wide capturing range (greater than 80%, from 10 2 to 10 5 cells ml −1 in phosphate-buffered saline buffer, pH 7.4). The platform showed a fast response time within a dynamic range of 10–10 5 cells ml −1 for the investigated T47D cancer cell line. We also obtained higher responses for anti-HER2 (human epidermal receptor 2)/streptavidin interface as the biosensing electrode in the presence of T47D cancer cells. The positive assay produced a sixfold increase in current output compared to the negative target or negative biological variant. We calculated the limit of detection at 0.4 U ml −1 , and of quantitation at 4 U ml −1 (units per millilitre). However, blood volume amounts in clinical settings may constrain diagnosis and increase detection limit value significantly.

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Electrochemical detection of protein glycosylation using lectin and protein–gold affinity interactions

Cited by 16 publications

References 28 publications

An electrochemical method for sensitive and rapid detection of FAM134B protein in colon cancer samples

An electrochemical method for sensitive and rapid detection of FAM134B protein in colon cancer samples

Gold-Loaded Nanoporous Ferric Oxide Nanocubes with Peroxidase-Mimicking Activity for Electrocatalytic and Colorimetric Detection of Autoantibody

Cancer biomarker profiling using nanozyme containing iron oxide loaded with gold particles

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