Application of Highly Sensitive Immunosensor Based on Optical Waveguide Light-Mode Spectroscopy (OWLS) Technique for...

The herbicide active ingredient glyphosate is the most widely applied herbicidal substance worldwide. Currently it is the market-leading pesticide, and its use is projected to further grow 4.5-fold between 2022 and 2029. Today, glyphosate use exceeds one megaton per year worldwide, which represents a serious environmental burden. A factor in the overall boost in the global use of glyphosate has been the spread of glyphosate-tolerant genetically modified (GM) crops that allow post-emergence applications of the herbicide on these transgenic crops. In turn, cultivation of glyphosate-tolerant GM crops represented 56% of the glyphosate use in 2019. Due to its extremely high application rate, xenobiotic behaviour and a water solubility (11.6 mg/mL at 25 °C) unusually high among pesticide active ingredients, glyphosate has become a ubiquitous water pollutant and a primary drinking water contaminant worldwide, presenting a threat to water quality. The goal of our research was to develop a rapid and sensitive method for detecting this herbicide active ingredient. For this purpose, we applied the novel analytical biosensor technique optical waveguide light-mode spectroscopy (OWLS) to the label-free detection of glyphosate in a competitive immunoassay format using glyphosate-specific polyclonal antibodies. After immobilising the antigen conjugate in the form of a glyphosate conjugated to human serum albumin for indirect measurement, the sensor chip was used in a flow-injection analyser system. For the measurements, an antibody stock solution was diluted to 2.5 µg/mL. During the measurement, standard solutions were mixed with the appropriate concentration of antibodies and incubated for 1 min before injection. The linear detection range and the EC₅₀ value of the competitive detection method were between 0.01 and 100 ng/mL and 0.60 ng/mL, respectively. After investigating the indirect method, we tested the cross-reactivity of the antibody with glyphosate and structurally related compounds.