ABSTRACT
After the COVID-19 pandemic started all over the world, great importance was placed on the development of sensitive and selective bioanalytical assays for the rapid detection of the highly pathogenic SARS-CoV-2 virus causing COVID-19 disease. In this present work, an impedimetric immunosensor was developed and applied for rapid, reliable, sensitive and selective detection of the SARS-CoV-2 S1 protein. To detect the SARS-CoV-2 virus, targeting of the spike S1 protein was achieved herein by using S1 protein-specific capture antibody (Cab-S1) immobilized screen-printed electrode (SPE) in combination with the electrochemical impedance spectroscopy (EIS) technique. With the impedimetric immunosensor, the detection limit for S1 protein in buffer medium was found to be 0.23 ng/mL (equal to 23.92 amol in 8 µL sample) in the linear concentration range of S1 protein from 0.5 to 10 ng/mL. In the artificial saliva medium, it was found to be 0.09 ng/mL (equals to 9.36 amol in 8 µL sample) in the linear concentration range of S1 protein between 0.1 and 1 ng/mL. The selectivity of the impedimetric immunosensor toward S1 protein was tested against influenza hemagglutinin antigen (HA) in the buffer medium as well as in artificial saliva.
ABSTRACT
In this present study, an amperometric immunosensor was developed based on disposable screen-printed carbon electrode (SPCE) for specific and sensitive detection of SARS-CoV-2 S1 protein. Anti-SARS-CoV-2 S1 monoclonal antibody was firstly immobilized onto the electrode surface. Then, the sandwich complex was formed by addition of S1 protein, secondary antibody and HRP-IgG, respectively. Chronoamperometry measurements were done in the presence of TMB mediator and the detection of SARS-CoV-2 S1 protein was performed by using 10 µL sample. The limit of detection (LOD) was found to be 0.19 ng/mL (equals to 24.7 amol in 10 µL sample) in the linear range of 0.5-10 ng/mL obtained in buffer medium. The applicability of this assay was investigated in the linear range of 0.5-3 ng/mL S1 protein in artificial saliva medium with the LOD as 0.13 ng/mL (equals to 16.9 amol in 10 µL sample). The selectivity study was examined in the presence of Hemagglutinin antigen (HA) in both mediums; buffer and artificial saliva while resulting with the successful discrimination between S1 protein and HA. The one of ultimate goals of our study is to present the possible implementation of this assay to point of care (POC) analysis. Under this aim, this assay was performed in combination with a portable device that is the commercial electrochemical analyzer. Amperometric detection of S1 protein in the range of 0.5-5 ng/mL was also successfully performed in artificial saliva medium with a resulting LOD as 0.15 ng/mL (equals to 19.5 amol in 10 µL sample). In addition, a selectivity study was similarly carried out by portable device.