An ELISA-Based Screening Platform for Ligand–Receptor Discovery

Sinem Ozgul, Sventja von Daake, Sumie Kakehi, Davide Sereni, Natalia Denissova, Carlie Hanlon, Yuanpeng Janet Huang, John K. Everett, Cuifeng Yin, Gaetano T. Montelione, Davide Comoletti

Research output: Chapter in Book/Report/Conference proceedingChapter

4 Scopus citations

Abstract

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.

Original languageEnglish (US)
Title of host publicationMethods in Enzymology
EditorsA. Joshua Wand
PublisherAcademic Press Inc.
Pages453-475
Number of pages23
ISBN (Print)9780128167625
DOIs
StatePublished - Jan 1 2019

Publication series

NameMethods in Enzymology
Volume615
ISSN (Print)0076-6879
ISSN (Electronic)1557-7988

All Science Journal Classification (ASJC) codes

  • Biochemistry
  • Molecular Biology

Keywords

  • Gene synthesis
  • Glycosylation
  • HEK cells
  • Mammalian cell surface proteins
  • NMR spectroscopy
  • Protein–protein interaction screen
  • Small-angle X-ray scattering
  • X-ray crystallography

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    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. In A. J. Wand (Ed.), Methods in Enzymology (pp. 453-475). (Methods in Enzymology; Vol. 615). Academic Press Inc.. https://doi.org/10.1016/bs.mie.2018.10.001