title: Antibody Drug-Target Engagement Measurement in Tissue Using Quantitative Affinity Extraction Liquid Chromatography-Mass Spectrometry: Method Development and Qualification.
abstract: We demonstrate a novel strategy using affinity extraction (AE) LC-MS to directly measure drug exposure and target engagement, two critical pharmacological questions, with a single assay. The assay measures total drug and target concentration at the site of therapeutic action, as well as the amount of target bound to drug. The case study presented applies the strategy to measure drug engagement of a membrane bound receptor (CD40) that is critical to immune regulation in colon biopsies collected from monkey dosed with an anti-CD40 antibody. Unlike other techniques that measure receptor occupancy, such as flow cytometry, this technique does not rely on viable cells allowing measurement of frozen samples in a remote setting from the clinic.
authors.affiliation: Bristol-Myers Squibb , Princeton, New Jersey 08543, United States.
authors.forename: Eugene
authors.identifier: 0000-0002-1334-1950
authors.initials: E
authors.lastname: Ciccimaro
authors.affiliation: Bristol-Myers Squibb , Princeton, New Jersey 08543, United States.
authors.forename: Yongxin
authors.initials: Y
authors.lastname: Zhu
authors.affiliation: Bristol-Myers Squibb , Princeton, New Jersey 08543, United States.
authors.forename: Dmitry
authors.initials: D
authors.lastname: Ostanin
authors.affiliation: Bristol-Myers Squibb , Princeton, New Jersey 08543, United States.
authors.forename: Suzanne
authors.initials: S
authors.lastname: Suchard
authors.affiliation: Bristol-Myers Squibb , Princeton, New Jersey 08543, United States.
authors.forename: Jamus
authors.initials: J
authors.lastname: MacGuire
authors.affiliation: Bristol-Myers Squibb , Princeton, New Jersey 08543, United States.
authors.forename: Qing
authors.initials: Q
authors.lastname: Xiao
authors.affiliation: Bristol-Myers Squibb , Princeton, New Jersey 08543, United States.
authors.forename: Ashok
authors.initials: A
authors.lastname: Dongre
authors.affiliation: Bristol-Myers Squibb , Princeton, New Jersey 08543, United States.
authors.forename: Anjaneya
authors.initials: A
authors.lastname: Chimalakonda
authors.affiliation: Bristol-Myers Squibb , Princeton, New Jersey 08543, United States.
authors.forename: Timothy
authors.initials: T
authors.lastname: Olah
authors.affiliation: Bristol-Myers Squibb , Princeton, New Jersey 08543, United States.
authors.forename: Petia
authors.initials: P
authors.lastname: Shipkova
mesh_terms: D000818:Animals; D000906:Antibodies / Q000032:analysis* / Q000276:immunology; D019013:CD40 Antigens / Q000032:analysis* / Q000276:immunology; D002846:Chromatography, Affinity / Q000379:methods; D003106:Colon / Q000737:chemistry*; D006801:Humans; D008252:Macaca fascicularis; D009092:Mucous Membrane / Q000737:chemistry*; D051381:Rats; D053719:Tandem Mass Spectrometry / Q000379:methods
chemical_list: D000906:Antibodies; D019013:CD40 Antigens