ABSTRACT
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.
Subject(s)
Biosensing Techniques , COVID-19 , Graphite , Humans , SARS-CoV-2 , Carbon , COVID-19/diagnosis , Biosensing Techniques/methods , Pandemics , Immunoassay/methods , Nucleocapsid , Electrodes , Antibodies, ViralABSTRACT
TiO2 time-dependent electrodeposited thin films were synthesized using an electrophoretic apparatus. The XRD analysis revealed that the films could exhibit a crystalline structure composed of ~81% anatase and ~6% rutile after 10 s of deposition, with crystallite size of 15 nm. AFM 3D maps showed that the surfaces obtained between 2 and 10 s of deposition exhibit strong topographical irregularities with long-range and short-range correlations being observed in different surface regions, a trend also observed by the Minkowski functionals. The height-based ISO, as well as specific surface microtexture parameters, showed an overall decrease from 2 to 10 s of deposition, showing a subtle decrease in the vertical growth of the films. The surfaces were also mapped to have low spatial dominant frequencies, which is associated with the similar roughness profile of the films, despite the overall difference in vertical growth observed. The electrical conductivity measurements showed that despite the decrease in topographical roughness, the films acquired a thickness capable of making them increasingly insulating from 2 to 10 s of deposition. Thus, our results prove that the deposition time used during the electrophoretic experiment consistently affects the films' structure, morphology, and electrical conductivity.
ABSTRACT
Molecularly imprinted polymers provide an excellent platform for the modification of selective electrodes for sensing applications. Herein, we present a novel modified carbon paste electrode (CPE) with a selective molecularly imprinted polymer (MIP) for recognition of sesquiterpene ß-caryophyllene, constituted of important plants oil-resins and extracts. The non-covalent MIP was synthesized using AA, EGDMA, and AIBN as a functional monomer, cross-linker and initiator agent, respectively. Structural and chemical characterization of the synthesized MIP was conducted through scanning electron microscopy (SEM), Fourier-transform infrared (FT-IR) spectroscopy, Raman spectroscopy and X-ray photoelectron spectroscopy (XPS). It was possible to verify the functional features of the synthesized MIP related to the extraction process of the template molecule. The CPE modified with MIP for sesquiterpene ß-caryophyllene recognition was characterized by electrochemical techniques as cyclic voltammetry (CV) and square wave voltammetry (SWV). The highest selective recognition electrode enables to detect concentrations in the range between 1.5â¯×â¯10-7 and 7.5â¯×â¯10-7â¯M, showing great potential for applications in monitoring content of sesquiterpene ß-caryophyllene in technological processes and for predicting the quality of extracts, oils, and resins of plants.