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 modernise 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 unlabelled 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 were well received by students, with most students able to follow the protocol and successfully determine ligand affinities. Furthermore, students recognised the benefit of the practical class for their education, confirming they felt it improved their understanding on how ligand affinity is calculated. We also demonstrated that this protocol could be expanded to large class sizes (class of 367 students studying medicine and veterinary sciences).