title: An ELISA Based Binding and Competition Method to Rapidly Determine Ligand-receptor Interactions.
abstract: A comprehensive understanding of signaling pathways requires detailed knowledge regarding ligand-receptor interaction. This article describes two fast and reliable point-by-point protocols of enzyme-linked immunosorbent assays (ELISAs) for the investigation of ligand-receptor interactions: the direct ligand-receptor interaction assay (LRA) and the competition LRA. As a case study, the ELISA based analysis of the interaction between different lambda interferons (IFNLs) and the alpha subunit of their receptor (IL28RA) is presented: the direct LRA is used for the determination of dissociation constants (KD values) between receptor and IFN ligands, and the competition LRA for the determination of the inhibitory capacity of an oligopeptide, which was designed to compete with the IFNLs at their receptor binding site. Analytical steps to estimate KD and half maximal inhibitory concentration (IC50) values are described. Finally, the discussion highlights advantages and disadvantages of the presented method and how the results enable a better molecular understanding of ligand-receptor interactions.
authors.affiliation: Applied Microbiology Research, Department of Biomedicine, University of Basel.
authors.forename: Mohameedyaseen
authors.initials: M
authors.lastname: Syedbasha
authors.affiliation: Applied Microbiology Research, Department of Biomedicine, University of Basel; Department of Biosystems Science and Engineering, ETH Zurich, and Swiss Institute of Bioinformatics; Swiss Institute of Bioinformatics.
authors.forename: Janina
authors.initials: J
authors.lastname: Linnik
authors.affiliation: Li Ka Shing Institute for Virology, University of Alberta.
authors.forename: Deanna
authors.initials: D
authors.lastname: Santer
authors.affiliation: Regional Infectious Diseases Unit, University of Edinburgh.
authors.forename: Daire
authors.initials: D
authors.lastname: O'Shea
authors.affiliation: Li Ka Shing Institute for Virology, University of Alberta; Faculty of Pharmacy and Pharmaceutical Sciences, University of Alberta.
authors.forename: Khaled
authors.initials: K
authors.lastname: Barakat
authors.affiliation: Li Ka Shing Institute for Virology, University of Alberta.
authors.forename: Michael
authors.initials: M
authors.lastname: Joyce
authors.affiliation: Infection Biology, Department of Biomedicine, University of Basel.
authors.forename: Nina
authors.initials: N
authors.lastname: Khanna
authors.affiliation: Li Ka Shing Institute for Virology, University of Alberta.
authors.forename: D Lorne
authors.initials: DL
authors.lastname: Tyrrell
authors.affiliation: Li Ka Shing Institute for Virology, University of Alberta.
authors.forename: Michael
authors.initials: M
authors.lastname: Houghton
authors.affiliation: Applied Microbiology Research, Department of Biomedicine, University of Basel; Clinical Microbiology, University Hospital Basel; adrian.egli@usb.ch.
authors.forename: Adrian
authors.initials: A
authors.lastname: Egli
mesh_terms: D001665:Binding Sites; D001667:Binding, Competitive / Q000502:physiology*; D055592:Biophysical Phenomena; D002352:Carrier Proteins; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods*; D007372:Interferons; D007378:Interleukins / Q000378:metabolism*; D008024:Ligands; D011485:Protein Binding / Q000502:physiology*; D018121:Receptors, Cytokine / Q000378:metabolism*; D017471:Receptors, Interferon; D000095022:Interferon Lambda
chemical_list: D002352:Carrier Proteins; D007372:Interferons; D007378:Interleukins; D008024:Ligands; D018121:Receptors, Cytokine; D017471:Receptors, Interferon; C000624353:interferon-lambda, human; D000095022:Interferon Lambda; C579640:IFNLR1 protein, human