Generic Automated Method for Liquid Chromatography鈥揗ultiple Reaction Monitoring Mass Spectrometry Based Monoclonal Antibody Quantitation for Preclinical Pharmacokinetic Studies

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• 作者：Qian Zhang ; Daniel S. Spellman ; Yaoli Song ; Bernard Choi ; Nathan G. Hatcher ; Daniela Tomazela ; Maribel Beaumont ; Mohammad Tabrizifard ; Deepa Prabhavalkar ; Wolfgang Seghezzi ; Jane Harrelson ; Kevin P. Bateman
• 刊名：Analytical Chemistry
• 出版年：2014
• 出版时间：September 2, 2014
• 年：2014
• 卷：86
• 期：17
• 页码：8776-8784
• 全文大小：407K
• ISSN：1520-6882

文摘

Quantitation of therapeutic monoclonal antibodies (mAb) using liquid chromatography鈥搕andem mass spectrometry (LC鈥揗S/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鈥揗S/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鈥揗S/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.