10.1021/acsomega.1c01959

Escherichia coli is a harmful Gram-negative bacterium commonly found in the gut of warm-blooded organisms and affects millions of people annually worldwide. In this study, we have synthesized a ZnO-CuO nanocomposite (NC) by a co-precipitation method and characterized the as-synthesized NC using FTIR spectroscopy, XRD, Raman spectroscopy, and FESEM techniques. To fabricate the immunosensor, the ZnO-CuO NC composite was screen-printed on gold-plated electrodes followed by physisorption of the anti-LPS E. coli antibody. The biosensor was optimized for higher specificity and sensitivity. The immunosensor exhibited a high sensitivity (11.04 μA CFU mL-1) with a low detection limit of 2 CFU mL-1 with a redox couple. The improved performance of the immunosensor is attributed to the synergistic effect of the NC and the antilipopolysaccharide antibody against E. coli. The selectivity studies were also carried out with Staphylococcus aureus to assess the specificity of the immunosensor. Testing in milk samples was done by spiking the milk samples with different concentrations of E. coli to check the potential of this immunosensor. We further checked the affinity between ZnO-CuO NC with E. coli LPS and the anti-LPS antibody using molecular docking studies. Atomic charge computation and interaction analyses were performed to support our hypothesis. Our results discern that there is a strong correlation between molecular docking studies and electrochemical characterization. The interaction analysis further displays the strong affinity between the antibody-LPS complex when immobilized with a nanoparticle composite (ZnO-CuO).