A Customizable Nanobody-Based Biosensor For Real-Time Pesticide Detection

ACustomizableNanobody-BasedBiosensorForReal-TimePesticideDetection Project Summary PD: Csicsery Nicholas Pesticides are among the most ubiquitous and persistent contaminants and human epidemio- logical studies have found associations between pesticide exposure and a number of adverse health conditions. We will develop a nanobody-based biosensor strain along with a on-line biosensor plat- form that uses a microfluidic device to detect and quantify the pesticide chlorpyrifos in real time. Objective1: Toidentifyandcharacterizethebindingkineticsofnanobodiesforchlorpyrifos. We will identify nanobodies that bind chlorpyrifos molecules and characterize their capture potential when surface displayed in an E. coli strain. We will work with a set of nanobodies that are enriched for specificity to the target and we will clone them into our E. coli nanobody display vector and screen and characterize them within our multiplexed microfluidic platform. Objective 2: To develop a prototype for continuous and batch chlorpyrifos sensing. We will develop an assay for measuring agglutination on a microfluidic-scale from many individual strain banks. We will optimize an agglutination assay using the surface-displayed nanobody strains and then transduce the agglutination signal using a customized computational algorithm. Finally we will optimize a sensor device to facilitate this assay and maximize the cellular signal so that we can quantify the amount of contaminant present in the water. Importantly this project serves as a seed project as once we have demonstrated proof of princi- ple with the development of a sensor for chlorpyrifos detection we will have a pipeline to rapidly develop sensors for many other emerging contaminants of concern. 1