Using a NanoBRET-Based Ligand-Binding Assay at the β₂-Adrenoceptor for Undergraduate Pharmacology Education

The teaching of pharmacology includes a fundamental understanding of ligand binding; how it is measured and how it is calculated. We sought to modernize the techniques by which ligand affinity is determined by undergraduate students and introduce them to parameter estimation through curve fitting. We taught a NanoBRET ligand binding assay to student cohorts completing their second year of undergraduate study in Natural Sciences (71 and 61 students in 2024 and 2025 respectively). The aim was to measure the affinity of fluorescent and unlabeled ligands for the β₂-adrenoceptor in live cells. Affinities were then calculated from their data using a custom Microsoft Excel spreadsheet. This series of practical classes was well received by students, with most students able to follow the protocol and successfully determine ligand affinities. Furthermore, students recognized the benefit of the practical class for their education, confirming they felt it improved their understanding of how ligand affinity is calculated. We also demonstrated that this protocol could be scaled up to accommodate larger class sizes (class of 367 students studying medicine and veterinary sciences).