Development of MS Binding Assays targeting the binding site of MB327 at the nicotinic acetylcholine receptor.

•MS Binding Assays for MB327, a potential nAChR-resensitizer, were developed.For quantification of MB327 a fast and sensitive LC-ESI-MS/MS method was established and validated.•MB327 binding at the nAChR was characterized for the first time.•MB327 binding at the nAChR was characterized for the first time.•These MS Binding Assays are the first tool for development of structure affinity relationships at the MB327 binding site.