title: Quantitative MALDI-MS Binding Assays: An Alternative to Radiolabeling.
abstract: Radiolabeling of ligands is still the gold standard in the study of high-affinity receptor-ligand interactions. In an effort toward safer and simpler alternatives to the use of radioisotopes, we developed a quantitative and highly sensitive matrix-assisted laser desorption ionization mass spectrometry (MALDI-MS) method that relies on the use of chemically tagged ligands designed to be specifically detectable when present as traces in complex biological mixtures such as cellular lysates. This innovative technology allows easy, sensitive detection and accurate quantification of analytes at the sub-nanomolar level. After statistical validation, we were able to perform pharmacological evaluations of G protein-coupled receptor (V1A-R)-ligand interactions. Both saturation and competitive binding assays were successfully processed.
authors.affiliation: Institut des Biomolécules Max Mousseron (IBMM), UMR5247 CNRS, ENSCM, Université de Montpellier, Place E. Bataillon, 34095, Montpellier Cedex 5, France.
authors.forename: Maxime
authors.initials: M
authors.lastname: Rossato
authors.affiliation: Institut des Biomolécules Max Mousseron (IBMM), UMR5247 CNRS, ENSCM, Université de Montpellier, Place E. Bataillon, 34095, Montpellier Cedex 5, France.
authors.forename: Guillaume
authors.initials: G
authors.lastname: Miralles
authors.affiliation: Institut des Biomolécules Max Mousseron (IBMM), UMR5247 CNRS, ENSCM, Université de Montpellier, Place E. Bataillon, 34095, Montpellier Cedex 5, France.
authors.forename: Céline
authors.initials: C
authors.lastname: M'Kadmi
authors.affiliation: Institut des Biomolécules Max Mousseron (IBMM), UMR5247 CNRS, ENSCM, Université de Montpellier, Place E. Bataillon, 34095, Montpellier Cedex 5, France.
authors.forename: Mathieu
authors.initials: M
authors.lastname: Maingot
authors.affiliation: Institut des Biomolécules Max Mousseron (IBMM), UMR5247 CNRS, ENSCM, Université de Montpellier, Place E. Bataillon, 34095, Montpellier Cedex 5, France.
authors.forename: Muriel
authors.initials: M
authors.lastname: Amblard
authors.affiliation: Institut de Génomique Fonctionnelle (IGF), 141 Rue de la Cardonille, 34090, Montpellier, France.
authors.forename: Bernard
authors.initials: B
authors.lastname: Mouillac
authors.affiliation: Institut des Biomolécules Max Mousseron (IBMM), UMR5247 CNRS, ENSCM, Université de Montpellier, Place E. Bataillon, 34095, Montpellier Cedex 5, France.
authors.forename: Didier
authors.initials: D
authors.lastname: Gagne
authors.affiliation: Institut des Biomolécules Max Mousseron (IBMM), UMR5247 CNRS, ENSCM, Université de Montpellier, Place E. Bataillon, 34095, Montpellier Cedex 5, France.
authors.forename: Jean
authors.initials: J
authors.lastname: Martinez
authors.affiliation: Institut des Biomolécules Max Mousseron (IBMM), UMR5247 CNRS, ENSCM, Université de Montpellier, Place E. Bataillon, 34095, Montpellier Cedex 5, France.
authors.forename: Gilles
authors.initials: G
authors.lastname: Subra
authors.affiliation: Institut des Biomolécules Max Mousseron (IBMM), UMR5247 CNRS, ENSCM, Université de Montpellier, Place E. Bataillon, 34095, Montpellier Cedex 5, France.
authors.forename: Christine
authors.initials: C
authors.lastname: Enjalbal
authors.affiliation: Institut des Biomolécules Max Mousseron (IBMM), UMR5247 CNRS, ENSCM, Université de Montpellier, Place E. Bataillon, 34095, Montpellier Cedex 5, France.
authors.forename: Sonia
authors.identifier: http://orcid.org/0000-0002-7583-8591
authors.initials: S
authors.lastname: Cantel
mesh_terms: D000818:Animals; D001667:Binding, Competitive; D016466:CHO Cells; D002623:Chemistry Techniques, Analytical / Q000379:methods*; D003373:Coumaric Acids / Q000737:chemistry / Q000378:metabolism; D006224:Cricetinae; D003412:Cricetulus; D007553:Isotope Labeling; D008024:Ligands; D010455:Peptides / Q000138:chemical synthesis / Q000737:chemistry; D043562:Receptors, G-Protein-Coupled / Q000378:metabolism; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization
chemical_list: D003373:Coumaric Acids; D008024:Ligands; D010455:Peptides; D043562:Receptors, G-Protein-Coupled
keywords: CHCA labeling; MALDI mass spectrometry; MS binding assays; peptide ligands; quantification; stable isotope dilution