A NanoBiT assay to monitor membrane proteins trafficking for drug discovery and drug development

Abstract

Internalization of membrane proteins plays a key role in many physiological functions; however, highly sensitive and versatile technologies are lacking to study such processes in real-time living systems. Here we describe an assay based on bioluminescence able to quantify membrane receptor trafficking for a wide variety of internalization mechanisms such as GPCR internalization/recycling, antibody-mediated internalization, and SARS-CoV2 viral infection. This study represents an alternative drug discovery tool to accelerate the drug development for a wide range of physiological processes, such as cancer, neurological, cardiopulmonary, metabolic, and infectious diseases including COVID-19. Membrane protein trafficking is monitored using split nanoluciferase. Receptor internalization leads to complementation on the early endosome and a bioluminescent response, and is applied to receptor internalization/recycling, antibody-mediated internalization and SARS-CoV2 entry.