An ultra-sensitive electrochemical biosensor using the Spike protein for capturing antibodies against SARS-CoV-2 in point-of-care. (December 2022)

• Record Type: Journal Article
• Authors: Cardoso, Ana R.; Alves, João Frederico; Frasco, Manuela F.; Piloto, Ana Margarida; Serrano, Verónica; Mateus, Daniela; Sebastião, Ana Isabel; Matos, Ana Miguel; Carmo, Anália; Cruz, Teresa; Fortunato, Elvira; Sales, M. Goreti F.
• Is Part Of: Materials today bio. Volume 16(2022)
• Publication Date: 2022-12
• Subjects: Electrochemical biosensor -- Antibodies for SARS-CoV-2 -- SARS-CoV-2 -- Spike protein -- Protective immunity -- Point-of-care; Materials science -- Periodicals; Biomedical engineering -- Periodicals; Biomedical materials -- Periodicals; 620.1
• DOI: 10.1016/j.mtbio.2022.100354
• Languages: English
• ISSNs: 2590-0064

Abstract

Abstract: This work presents an innovative ultra-sensitive biosensor having the Spike protein on carbon-based screen-printed electrodes (SPEs), for monitoring in point-of-care antibodies against SARS-CoV-2, a very important tool for epidemiological monitoring of COVID-19 infection and establishing vaccination schemes. In an innovative and simple approach, a highly conductive support is combined with the direct adsorption of Spike protein to enable an extensive antibody capture. The high conductivity was ensured by using carboxylated carbon nanotubes on the carbon electrode, by means of a simple and quick approach, which also increased the surface area. These were then modified with EDC/NHS chemistry to produce an amine layer and undergo Spike protein adsorption, to generate a stable layer capable of capturing the antibodies against SARS-CoV-2 in serum with great sensitivity. Electrochemical impedance spectroscopy was used to evaluate the analytical performance of this biosensor in serum. It displayed a linear response between 1.0 ​pg/mL and 10 ​ng/mL, with a detection limit of ∼0.7 ​pg/mL. The analysis of human positive sera containing antibody in a wide range of concentrations yielded accurate data, correlating well with the reference method. It also offered the unique ability of discriminating antibody concentrations in sera below 2.3 ​μg/mL, the lowest value detected by the commercial method. In addition, a proof-of-concept study was performed by labelling anti-IgG antibodies with quantum dots to explore a new electrochemical readout based on the signal generated upon binding to the anti-S protein antibodies recognised on the surface of the biosensor. Overall, the alternative serologic assay presented is a promising tool for assessing protective immunity to SARS-CoV-2 and a potential guide for revaccination. Graphical abstract: Image 1 Highlights: An ultra-sensitive biosensor for detection of low levels of antibodies against SARS-CoV-2. Highly conductive substrate with adsorbed protein S and point-of-care capability. Application to human sera samples and good correlation with commercial method. Electrochemical impedance readings with an iron-based redox probe. Alternative electrochemical impedance readings with anti-IgG labelled with quantum dots.