Application of frontal affinity chromatography with mass spectrometry (FAC-MS) for stereospecific ligand-macromolecule interaction, detection and screening.
Using frontal affinity chromatography coupled to mass spectrometry (FAC-MS) we have established a general stereoselective detection and screening method of intact racemates which can generate binding affinity information about the individual enantiomers that is also applicable to other ligand isomeric mixtures. FAC-MS has been shown to be a versatile technology utilizing direct binding in screening assays and extending its application toward chiral drug development, especially in the early discovery stages as well as its utility in secondary Structure-activity relationship (SAR) studies allow this platform to make a significant step toward facilitating the demand for pure enantiomeric drugs. Using renin, which is as an important drug target, we show that for detection and screening purposes there is no need to first use timely and costly methods of separating racemates in order to get precise information about the binding affinities of the composite enantiomers.
Slon-Usakiewicz
Jacek J
JJ
jusakiewicz@cogeco.ca
Redden
Peter
P
D002846:Chromatography, Affinity / Q000379:methods*; D004353:Drug Evaluation, Preclinical / Q000379:methods*; D006801:Humans; D055767:Immobilized Proteins / Q000737:chemistry / Q000378:metabolism; D008024:Ligands; D046911:Macromolecular Substances / Q000737:chemistry / Q000378:metabolism*; D013058:Mass Spectrometry / Q000379:methods*; D012083:Renin / Q000737:chemistry / Q000378:metabolism; D013237:Stereoisomerism; D013379:Substrate Specificity
D055767:Immobilized Proteins; D008024:Ligands; D046911:Macromolecular Substances; D012083:Renin