title: Techniques for quantitative LC-MS/MS analysis of protein therapeutics: advances in enzyme digestion and immunocapture.
abstract: LC-MS/MS has been investigated to quantify protein therapeutics in biological matrices. The protein therapeutics is digested by an enzyme to generate surrogate peptide(s) before LC-MS/MS analysis. One challenge is isolating protein therapeutics in the presence of large number of endogenous proteins in biological matrices. Immunocapture, in which a capture agent is used to preferentially bind the protein therapeutics over other proteins, is gaining traction. The protein therapeutics is eluted for digestion and LC-MS/MS analysis. One area of tremendous potential for immunocapture-LC-MS/MS is to obtain quantitative data where ligand-binding assay alone is not sufficient, for example, quantitation of antidrug antibody complexes. Herein, we present an overview of recent advance in enzyme digestion and immunocapture applicable to protein quantitation.
authors.affiliation: Research and Development, Bristol-Myers Squibb Company, 1 Squibb Drive, New Brunswick, NJ 08943, USA.
authors.forename: Eliza N
authors.initials: EN
authors.lastname: Fung
authors.affiliation: B2S Consulting, Mendham, NJ 07945, USA.
authors.forename: Peter
authors.initials: P
authors.lastname: Bryan
authors.affiliation: Research and Development, Bristol-Myers Squibb Company, Route 206/Province Line Road, Princeton, NJ 08543, USA.
authors.forename: Alexander
authors.initials: A
authors.lastname: Kozhich
mesh_terms: D002846:Chromatography, Affinity; D002851:Chromatography, High Pressure Liquid; D006801:Humans; D007074:Immunoglobulin G / Q000276:immunology; D010447:Peptide Hydrolases / Q000378:metabolism*; D011506:Proteins / Q000032:analysis* / Q000302:isolation & purification / Q000378:metabolism; D053719:Tandem Mass Spectrometry
chemical_list: D007074:Immunoglobulin G; D011506:Proteins; D010447:Peptide Hydrolases
keywords: LC–MS/MS; acid hydrolysis; enzyme digestion; immunocapture; protein quantitation; regulated bioanalysis