An Ultra-Rapid Biosensory Point-of-Care (POC) Assay for Prostate-Specific Antigen (PSA) Detection in Human Serum

Mavrikou, Sophie; Moschopoulou, Georgia; Zafeirakis, Athanasios; Kalogeropoulou, Konstantina; Giannakos, Georgios; Skevis, Athanasios; Kintzios, Spyridon

doi:10.3390/s18113834

Cited by 23 publications

(21 citation statements)

References 60 publications

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“…A 7 µL sample of human serum (1:200 v/v prepared in PBS solution) was dropped on the WE and incubated for 30 min. After washing with water, 7 µL of horseradish peroxidase-labeled anti-human IgG (anti-IgG-HRP) (1:1000 v/v (KPL Inc.) prepared in PBS solution) was added, and incubated for 30 min at room temperature [ 24 , 25 , 43 ]. Immediately, 100 µL of TMB was placed to promote the enzymatic reaction, and the chronoamperometric (CA) technique followed the reduction process by applying a constant potential of −0.19 V for 50 s ( Scheme 1 f–h) [ 44 , 45 ].…”

Section: Methodsmentioning

confidence: 99%

An Electrochemical Immunosensor Based on Carboxylated Graphene/SPCE for IgG-SARS-CoV-2 Nucleocapsid Determination

Freire

Ruzo

Salgado³

et al. 2022

Biosensors

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The COVID-19 pandemic has emphasized the importance and urgent need for rapid and accurate diagnostic tests for detecting and screening this infection. Our proposal was to develop a biosensor based on an ELISA immunoassay for monitoring antibodies against SARS-CoV-2 in human serum samples. The nucleocapsid protein (N protein) from SARS-CoV-2 was employed as a specific receptor for the detection of SARS-CoV-2 nucleocapsid immunoglobulin G. N protein was immobilized on the surface of a screen-printed carbon electrode (SPCE) modified with carboxylated graphene (CG). The percentage of IgG-SARS-CoV-2 nucleocapsid present was quantified using a secondary antibody labeled with horseradish peroxidase (HRP) (anti-IgG-HRP) catalyzed using 3,3′,5,5′-tetramethylbenzidine (TMB) mediator by chronoamperometry. A linear response was obtained in the range of 1:1000–1:200 v/v in phosphate buffer solution (PBS), and the detection limit calculated was 1:4947 v/v. The chronoamperometric method showed electrical signals directly proportional to antibody concentrations due to antigen-antibody (Ag-Ab) specific and stable binding reaction.

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Section: Methodsmentioning

confidence: 99%

An Electrochemical Immunosensor Based on Carboxylated Graphene/SPCE for IgG-SARS-CoV-2 Nucleocapsid Determination

Freire

Ruzo

Salgado³

et al. 2022

Biosensors

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show abstract

“…This possibility of inter-assay error is markedly reduced by combining electrochemistry on a microfluidic chip. Mavrikou et al [ 52 ] developed a biosensor whose membrane potential is modified by engineered cells in the presence of PSA, indicating a high (>4 ng mL −1 ) or low (<4 ng mL −1 ) PSA level. However, the use of cells as the sensing element limits the ease of use and the storage and portability of the device.…”

Section: Discussionmentioning

confidence: 99%

A Novel, Quick, and Reliable Smartphone-Based Method for Serum PSA Quantification: Original Design of a Portable Microfluidic Immunosensor-Based System

Ortega

Gómez

González-Martinez

et al. 2022

Cancers

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We describe a versatile, portable, and simple platform that includes a microfluidic electrochemical immunosensor for prostate-specific antigen (PSA) detection. It is based on the covalent immobilization of the anti-PSA monoclonal antibody on magnetic microbeads retained in the central channel of a microfluidic device. Image flow cytometry and scanning electron microscopy were used to characterize the magnetic microbeads. A direct sandwich immunoassay (with horseradish peroxidase-conjugated PSA antibody) served to quantify the cancer biomarker in serum samples. The enzymatic product was detected at −100 mV by amperometry on sputtered thin-film electrodes. Electrochemical reaction produced a current proportional to the PSA level, with a linear range from 10 pg mL−1 to 1500 pg mL−1. The sensitivity was demonstrated by a detection limit of 2 pg mL−1 and the reproducibility by a coefficient of variation of 6.16%. The clinical performance of this platform was tested in serum samples from patients with prostate cancer (PCa), observing high specificity and full correlation with gold standard determinations. In conclusion, this analytical platform is a promising tool for measuring PSA levels in patients with PCa, offering a high sensitivity and reduced variability. The small platform size and low cost of this quantitative methodology support its suitability for the fast and sensitive analysis of PSA and other circulating biomarkers in patients. Further research is warranted to verify these findings and explore its potential application at all healthcare levels.

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“…The detection of antibodies was carried out by enzymatic reaction using TMB as substrate for 1 min in the dark. Firstly, the human serum sample (1:200 v/v prepared in PBS solution) was incubated for 30 min on the WE, followed by the addition of horseradish peroxidase-labeled anti-human IgG (anti-IgG-HRP) (1:1000 v/v prepared in PBS solution) for another 30 min at room temperature [22][23][24]. The enzymatic reaction occurs when TMB is added after the previous step.…”

Section: Electrochemical Detectionmentioning

confidence: 99%

Electrochemical Immunosensor Based on Carboxylated Graphene/SPCE for Igg-SARS-Cov-2 Nucleocapsid Determination

Brito¹,

Freire²,

Ruzo³

et al. 2022

Preprint

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The COVID-19 pandemic has highlighted the importance and urgent need of rapid and accurate diagnostic tests for detection and screening of this infection. In our proposal, a biosensor based on the ELISA immunoassay was developed for monitoring antibodies against SARS-CoV-2 in human serum samples. The SARS-CoV-2 nucleocapsid protein (N-protein) was selected as a specific receptor for the detection of SARS-CoV-2 nucleocapsid immunoglobulin G. Thus, the N-protein was immobilized on surface of screen-printed carbon electrode (SPCE) modified with carboxylated graphene (CG). The IgG-SARS-CoV-2 nucleocapsid concentration was quantified using a secondary antibody labelled with horseradish peroxidase (HRP) (anti-IgG-HRP) catalyzed by 3,3’,5,5’-tetramethylbenzidine (TMB) mediator by chronoamperometry. A linear response was obtained in the range of 1:1000-1:200 v/v in phosphate buffer solution (PBS) and the limit of detection calculated was of 1:4947 v/v. The chronoamperometric method showed electrical signals directly proportional to antibody concentrations due to Ag-Ab specific and stable binding reaction.

show abstract

An Ultra-Rapid Biosensory Point-of-Care (POC) Assay for Prostate-Specific Antigen (PSA) Detection in Human Serum

Cited by 23 publications

References 60 publications

An Electrochemical Immunosensor Based on Carboxylated Graphene/SPCE for IgG-SARS-CoV-2 Nucleocapsid Determination

An Electrochemical Immunosensor Based on Carboxylated Graphene/SPCE for IgG-SARS-CoV-2 Nucleocapsid Determination

A Novel, Quick, and Reliable Smartphone-Based Method for Serum PSA Quantification: Original Design of a Portable Microfluidic Immunosensor-Based System

Electrochemical Immunosensor Based on Carboxylated Graphene/SPCE for Igg-SARS-Cov-2 Nucleocapsid Determination

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