title: Generic automated method for liquid chromatography-multiple reaction monitoring mass spectrometry based monoclonal antibody quantitation for preclinical pharmacokinetic studies.
abstract: Quantitation of therapeutic monoclonal antibodies (mAb) using liquid chromatography-tandem mass spectrometry (LC-MS/MS) for pharmacokinetic (PK) studies is becoming an essential complement to traditional antibody-based ligand binding assays (LBA). Here we show an automated method to perform LC-MS/MS-based quantitation, with IgG1 conserved peptides, a heavy isotope labeled mAb internal standard, and anti-human Fc enrichment. All reagents in the method are commercially available with no requirement to develop novel assay-specific reagents. The method met traditional quantitative LC-MS/MS assay analytical characteristics in terms of precision, accuracy, and specificity. The method was applied to the pharmacokinetic study of a mAb dosed in cynomolgus monkey, and the results were compared with the immunoassay data. This methodology has the potential to benefit and accelerate the early biopharmaceutical development process, particularly by enabling PK analysis across species and candidate molecules with minimal method development.
authors.affiliation: Pharmacokinetics, Pharmacodynamics and Drug Metabolism, Merck & Co., Incorporated , West Point, Pennsylvania 19486, United States.
authors.forename: Qian
authors.initials: Q
authors.lastname: Zhang
authors.forename: Daniel S
authors.initials: DS
authors.lastname: Spellman
authors.forename: Yaoli
authors.initials: Y
authors.lastname: Song
authors.forename: Bernard
authors.initials: B
authors.lastname: Choi
authors.forename: Nathan G
authors.initials: NG
authors.lastname: Hatcher
authors.forename: Daniela
authors.initials: D
authors.lastname: Tomazela
authors.forename: Maribel
authors.initials: M
authors.lastname: Beaumont
authors.forename: Mohammad
authors.initials: M
authors.lastname: Tabrizifard
authors.forename: Deepa
authors.initials: D
authors.lastname: Prabhavalkar
authors.forename: Wolfgang
authors.initials: W
authors.lastname: Seghezzi
authors.forename: Jane
authors.initials: J
authors.lastname: Harrelson
authors.forename: Kevin P
authors.initials: KP
authors.lastname: Bateman
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D000911:Antibodies, Monoclonal / Q000097:blood / Q000493:pharmacokinetics*; D001774:Blood Chemical Analysis / Q000295:instrumentation*; D002851:Chromatography, High Pressure Liquid; D004353:Drug Evaluation, Preclinical; D004797:Enzyme-Linked Immunosorbent Assay; D006207:Half-Life; D007074:Immunoglobulin G / Q000378:metabolism; D047468:Immunoprecipitation; D007553:Isotope Labeling; D008252:Macaca fascicularis; D008969:Molecular Sequence Data; D010455:Peptides / Q000032:analysis* / Q000737:chemistry; D053719:Tandem Mass Spectrometry
chemical_list: D000911:Antibodies, Monoclonal; D007074:Immunoglobulin G; D010455:Peptides