NanoB² to monitor interactions of ligands with membrane proteins by combining nanobodies and NanoBRET.

ABSTRACT: The therapeutic potential of ligands targeting disease-associated membrane proteins is predicted by ligand-receptor binding constants, which can be determined using NanoLuciferase (NanoLuc)-based bioluminescence resonance energy transfer (NanoBRET) methods. However, the broad applicability of these methods is hampered by the restricted availability of fluorescent probes. We describe the use of antibody fragments, like nanobodies, as universal building blocks for fluorescent probes for use in NanoBRET. Our nanobody-NanoBRET (NanoB²) workflow starts with the generation of NanoLuc-tagged receptors and fluorescent nanobodies, enabling homogeneous, real-time monitoring of nanobody-receptor binding. Moreover, NanoB² facilitates the assessment of receptor binding of unlabele

SUBMITTER: van den Bor J 

PROVIDER: S-EPMC10088090 | biostudies-literature | 2023 Mar

REPOSITORIES: biostudies-literature