Premium
Quantitative MALDI‐MS Binding Assays: An Alternative to Radiolabeling
Author(s) -
Rossato Maxime,
Miralles Guillaume,
M'Kadmi Céline,
Maingot Mathieu,
Amblard Muriel,
Mouillac Bernard,
Gagne Didier,
Martinez Jean,
Subra Gilles,
Enjalbal Christine,
Cantel Sonia
Publication year - 2016
Publication title -
chemmedchem
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.817
H-Index - 100
eISSN - 1860-7187
pISSN - 1860-7179
DOI - 10.1002/cmdc.201600447
Subject(s) - chemistry , analyte , mass spectrometry , combinatorial chemistry , ligand (biochemistry) , matrix assisted laser desorption/ionization , competitive binding , chromatography , computational biology , receptor , desorption , biochemistry , organic chemistry , adsorption , biology
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.