title: Strategies for improving sensitivity and selectivity for the quantitation of biotherapeutics in biological matrix using LC-MS/MS.
abstract: In recent years, the applicability of using LC-MS/MS as a complementary technique to traditional ligand binding assays in the absolute quantitation of therapeutic proteins in biologic matrix has been demonstrated. Protein quantitation workflow via LC-MS/MS is primarily based on a enzymatic digestion model and recent works seek to improve selectivity and sensitivity. This review focuses on recent innovations in this field and discusses the following in detail: the applicability of two-dimensional liquid chromatography and its use to improve sensitivity and alleviate matrix ion suppression; the use of derivatization agents after digestion to improve extraction and MS ionization efficiency; techniques to reduce excess protein background and their positive effects on sensitivity, selectivity, and extraction consistency; the application of immunoaffinity extraction of proteins to enrich the analyte(s) of interest while improving selectivity and sensitivity.
authors.affiliation: Bristol-Myers Squibb - Analytical and Bioanalytical Development, Route 206 and Province Line Road, Princeton, NJ 08502, USA.
authors.forename: Jim X
authors.initials: JX
authors.lastname: Shen
authors.forename: Guowen
authors.initials: G
authors.lastname: Liu
authors.forename: Yue
authors.initials: Y
authors.lastname: Zhao
mesh_terms: D000818:Animals; D001691:Biological Therapy / Q000379:methods; D002853:Chromatography, Liquid / Q000379:methods; D006801:Humans; D013058:Mass Spectrometry / Q000379:methods; D011506:Proteins / Q000737:chemistry*; D012680:Sensitivity and Specificity
chemical_list: D011506:Proteins
keywords: LC-MS/MS; bioanalytical; biotherapeutics; enzymatic digestion; quantitation; selectivity; sensitivity