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Updated: Oct 8, 2025

Measuring G-protein-coupled Receptor Signaling via Radio-labeled GTP Binding
Published on: June 9, 2017
Activation of a G protein-coupled receptor through indirect antibody-mediated tethering of ligands
Ross W Cheloha1, Fabian A Fischer1, Thomas J Gardella2
1Program in Cellular and Molecular Medicine, Boston Children's Hospital, 1 Blackfan Circle Boston MA 02115 USA ross.cheloha@nih.gov hidde.ploegh@childrens.harvard.edu.
Abstract:
Antibodies raised against many cell surface proteins, including G protein-coupled receptors, remain important tools for their functional characterization. By linking antibodies to ligands for cell surface proteins, such adducts can be targeted to the surface of a cell type of choice. Site-specific functionalization of full-size antibodies with synthetic moieties remains challenging. Here we present new approaches in which single domain antibodies (known as VHHs or nanobodies) that target either cell surface proteins or conventional antibodies are used to indirectly deliver ligands for GPCRs to their sites of action. The combination of high yield production of nanobodies, facile site-specific functionalization, and compatibility with commercially available mouse and rabbit antibodies should enable wide application of this approach.
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