NanoResponder – a bacterial two hybrid selection system to generate custom single-domain antibodies

The prevalence of virulent and multiple antibiotic resistant pathogens in healthcare facilities has resulted in the ongoing reassessment of best practices to prevent their transmission. However, whether pathogen reservoirs exist in emergency medical services (EMS) vehicles remains largely unknown. We characterized the microbiome of EMS vehicles using Nanopore next generation sequencing of isolated DNA samples. Based on the pathogens identified during sequencing, we developed a bacterial two hybrid selection system to generate custom single-domain antibodies (Nanobodies™) that recognize important pathogens. Selected antibodies were then used to create a rapid, low-cost and on site ELISA-based testing kit. Our project provides a framework for the detection of emergent pathogens and a practical solution for monitoring their presence within and outside of the healthcare system.