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Selection of Transporter-Targeted Inhibitory Nanobodies by Solid-Supported-Membrane (SSM)-Based Electrophysiology
JournalArticle (Originalarbeit in einer wissenschaftlichen Zeitschrift)
 
ID 4619131
Author(s) Bärland, Natalie; Perez, Camilo
Author(s) at UniBasel Perez, Camilo
Bärland, Natalie
Year 2021
Title Selection of Transporter-Targeted Inhibitory Nanobodies by Solid-Supported-Membrane (SSM)-Based Electrophysiology
Journal Journal of visualized experiments
Volume 171
Number 171
Pages / Article-Number e62578
Mesh terms Cardiac Electrophysiology; Humans; Kinetics; Membrane Proteins, metabolism; Membrane Transport Proteins, metabolism; Single-Domain Antibodies
Abstract Single domain antibodies (nanobodies) have been extensively used in mechanistic and structural studies of proteins and they pose an enormous potential as tools for developing clinical therapies, many of which depend on the inhibition of membrane proteins such as transporters. However, most of the methods used to determine the inhibition of transport activity are difficult to perform in high-throughput routines and depend on labeled substrates availability thereby complicating the screening of large nanobody libraries. Solid-supported membrane (SSM) electrophysiology is a high-throughput method, used for characterizing electrogenic transporters and measuring their transport kinetics and inhibition. Here we show the implementation of SSM-based electrophysiology to select inhibitory and non-inhibitory nanobodies targeting an electrogenic secondary transporter and to calculate nanobodies inhibitory constants. This technique may be especially useful for selecting inhibitory nanobodies targeting transporters for which labeled substrates are not available.
Publisher JoVE
ISSN/ISBN 1940-087X
edoc-URL https://edoc.unibas.ch/82959/
Full Text on edoc No
Digital Object Identifier DOI 10.3791/62578
PubMed ID http://www.ncbi.nlm.nih.gov/pubmed/33999018
ISI-Number WOS:000647027000010
Document type (ISI) Journal Article
 
   

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