title: Ultrasensitive quantitative LC-MS/MS of an inhibitor of apoptosis protein's antagonist in plasma using protein target affinity extraction.
abstract: BACKGROUND
  A target protein-based affinity extraction LC-MS/MS method was developed to enable plasma level determination following ultralow dosing (0.1-3 µg/kg) of an inhibitor of apoptosis proteins molecule. Methodology & results: Affinity extraction (AE) utilizing immobilized target protein BIR2/BIR3 was used to selectively capture the inhibitor of apoptosis proteins molecule from dog plasma and enable removal of background matrix components. Pretreatment of plasma samples using protein precipitation was found to provide an additional sensitivity gain. A LLOQ of 7.8 pM was achieved by combining protein precipitation with AE. The method was used to support an ultralow dose dog toxicity study.
  
  
  CONCLUSIONS
  AE-LC-MS/MS, utilizing target protein, is a highly sensitive methodology for small molecule quantification with potential for broader applicability.
authors.affiliation: Bioanalytical Research, Bristol-Myers Squibb Company, Route 206 & Province Line Road, Princeton, NJ 08543, USA.
authors.forename: Lorell N
authors.initials: LN
authors.lastname: Discenza
authors.affiliation: Bioanalytical Research, Bristol-Myers Squibb Company, Route 206 & Province Line Road, Princeton, NJ 08543, USA.
authors.forename: Georgia
authors.initials: G
authors.lastname: Cornelius
authors.affiliation: Biotransformation, Bristol-Myers Squibb Company, Route 206 & Province Line Road, Princeton, NJ 08543, USA.
authors.forename: Jinping
authors.initials: J
authors.lastname: Gan
authors.affiliation: Protein Science, Bristol-Myers Squibb Company, Route 206 & Province Line Road, Princeton, NJ 08543, USA.
authors.forename: Nicolas
authors.initials: N
authors.lastname: Szapiel
authors.affiliation: Oncology Discovery, Bristol-Myers Squibb Company, Route 206 & Province Line Road, Princeton, NJ 08543, USA.
authors.forename: Randy L
authors.initials: RL
authors.lastname: Talbott
authors.affiliation: Mechanistic Biochemistry, Bristol-Myers Squibb Company, Route 206 & Province Line Road, Princeton, NJ 08543, USA.
authors.forename: Charu
authors.initials: C
authors.lastname: Chaudhry
authors.affiliation: Discovery Toxicology, Bristol-Myers Squibb Company, Route 206 & Province Line Road, Princeton, NJ 08543, USA.
authors.forename: Anirban
authors.initials: A
authors.lastname: Roy
authors.affiliation: Oncology Chemistry, Bristol-Myers Squibb Company, Route 206 & Province Line Road, Princeton, NJ 08543, USA.
authors.forename: Robert M
authors.initials: RM
authors.lastname: Borzilleri
authors.affiliation: Metabolism & Pharmacokinetics, Bristol-Myers Squibb Company, Route 206 & Province Line Road, Princeton, NJ 08543, USA.
authors.forename: Ragini
authors.initials: R
authors.lastname: Vuppugalla
authors.affiliation: Pharmaceutics, Bristol-Myers Squibb Company, Route 206 & Province Line Road, Princeton, NJ 08543, USA.
authors.forename: Kevin
authors.initials: K
authors.lastname: Stefanski
authors.affiliation: Metabolism & Pharmacokinetics, Bristol-Myers Squibb Company, Route 206 & Province Line Road, Princeton, NJ 08543, USA.
authors.forename: Robin
authors.initials: R
authors.lastname: Moore
authors.affiliation: Bioanalytical Research, Bristol-Myers Squibb Company, Route 206 & Province Line Road, Princeton, NJ 08543, USA.
authors.forename: Celia J
authors.initials: CJ
authors.lastname: D'Arienzo
authors.affiliation: Bioanalytical Research, Bristol-Myers Squibb Company, Route 206 & Province Line Road, Princeton, NJ 08543, USA.
authors.forename: Timothy V
authors.initials: TV
authors.lastname: Olah
authors.affiliation: Bioanalytical Research, Bristol-Myers Squibb Company, Route 206 & Province Line Road, Princeton, NJ 08543, USA.
authors.forename: John T
authors.initials: JT
authors.lastname: Mehl
mesh_terms: D000818:Animals; D001774:Blood Chemical Analysis / Q000379:methods*; D005591:Chemical Fractionation / Q000379:methods*; D002853:Chromatography, Liquid / Q000379:methods*; D004285:Dogs; D005260:Female; D006801:Humans; D055767:Immobilized Proteins / Q000037:antagonists & inhibitors / Q000737:chemistry; D051034:Inhibitor of Apoptosis Proteins / Q000037:antagonists & inhibitors* / Q000737:chemistry; D007546:Isoquinolines / Q000032:analysis* / Q000737:chemistry / Q000494:pharmacology; D057230:Limit of Detection; D008297:Male; D009842:Oligopeptides / Q000032:analysis* / Q000737:chemistry / Q000494:pharmacology; D054852:Small Molecule Libraries / Q000032:analysis* / Q000737:chemistry / Q000494:pharmacology; D053719:Tandem Mass Spectrometry / Q000379:methods*
chemical_list: C000611973:BMS-986152; D055767:Immobilized Proteins; D051034:Inhibitor of Apoptosis Proteins; D007546:Isoquinolines; D009842:Oligopeptides; D054852:Small Molecule Libraries
keywords: LC–MS/MS; MS; affinity extraction; affinity precipitation; enzyme inhibition constant (Ki); ligand-binding assay