A SARS-CoV-2 impedimetric biosensor based on the immobilization of ACE-2 receptor-containing entire cell membranes as the biorecognition element

Abstract

A SARS-CoV-2 biosensor based on the biorecognition of the spike protein to the angiotensin-converting enzyme 2 (ACE-2) transmembrane receptor was developed using entire cell membranes as the biorecognition layer. In this new SARS-CoV-2 detection platform, cellular membranes from VeroCCL81 (mVero) and Calu-3 (mCalu) cells (which overexpress the ACE-2 transmembrane receptors) were extracted and immobilized as vesicles on an indium tin oxide electrode (ITO). Electrochemical impedance spectroscopy was used to optimize the performance of the developed devices for SARS-CoV-2 detection. This novel biosensor comprises a low-cost system (less than one US$ dollar) that uses the unique properties of cell membranes combined with the catalytic properties of electrochemical platforms to allow spike proteins recognition. A linear response from 10 to 100 ng/mL was obtained from the optimized biosensors, a limit of detection of 10.0 pg/mL and 7.25 pg/mL and limit of quantification of 30.4 pg/mL and 21.9 pg/mL were achieved with satisfactory accuracy for ITO-APTES-mVero and ITO-APTES-mCalu, respectively. Selectivity studies revealed that this platform was able to differentiate the target spike proteins from NS1 proteins from dengue and Zika viruses. In addition, sensors comprising cell membranes devoid of the ACE-2 transmembrane receptor exhibited no biorecognition signal. The developed devices are suitable for SARS-CoV-2 detection based on spike protein recognition, and capable of providing a low-cost, accurate, and accessible tool for use in a pandemic and post-pandemic scenario.