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An ELISA-Based Screening Platform for Ligand–Receptor Discovery

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posted on 2024-02-22, 03:26 authored by S Ozgul, S von Daake, S Kakehi, D Sereni, N Denissova, C Hanlon, YJ Huang, JK Everett, C Yin, GT Montelione, Davide ComolettiDavide Comoletti
Cell surface molecules are important for development and function of multicellular organisms. Although several methods are available to identify ligand–receptor pairs, ELISA-based methods are particularly amenable to high-throughput screens. ELISA-based methods have high sensitivity and low false-positive rates for detecting protein–protein interaction (PPI) complexes. Here, we provide a detailed protocol for a 384-well ELISA-based PPI screening protocol for the identification of novel cell surface ligand–receptor interactions, together with considerations for validation of PPIs by biophysical methods. This PPI screen has been developed and tested for discovery of novel ligand–receptor pairs between human synaptic adhesion proteins, believed to play crucial roles in many steps of neurodevelopment, from neuronal maturation, to axon guidance, synapse connectivity, and pruning.

History

Preferred citation

Ozgul, S., von Daake, S., Kakehi, S., Sereni, D., Denissova, N., Hanlon, C., Huang, Y. J., Everett, J. K., Yin, C., Montelione, G. T. & Comoletti, D. (2019). An ELISA-Based Screening Platform for Ligand–Receptor Discovery. Methods in Enzymology (615, pp. 453-475). Elsevier. https://doi.org/10.1016/bs.mie.2018.10.001

Book title

Methods in Enzymology

Publisher

Elsevier

Pagination

453-475

Series

Methods in Enzymology

Volume

615

ISBN

9780128167625

ISSN

0076-6879