title: Multiple heart-cutting and comprehensive two-dimensional liquid chromatography hyphenated to mass spectrometry for the characterization of the antibody-drug conjugate ado-trastuzumab emtansine.
abstract: Antibody-drug conjugates might be the magic bullets referred to by Paul Ehrlich over 100 years ago. Together with a huge therapeutic potential, these molecules come with a structural complexity that drives state-of-the-art chromatography and mass spectrometry to its limits. The use of multiple heart-cutting (mLC-LC) and comprehensive (LC×LC) multidimensional LC in combination with high resolution mass spectrometry for the characterization of the lysine conjugated antibody-drug conjugate ado-trastuzumab emtansine, commercialized as Kadcyla, is presented. By combining protein and peptide measurements, attributes such as drug loading, drug distribution and drug conjugation sites can be assessed in an elegant manner.
authors.affiliation: Research Institute for Chromatography, President Kennedypak 26, B-8500 Kortrijk, Belgium. Electronic address: koen.sandra@richrom.com.
authors.forename: Koen
authors.initials: K
authors.lastname: Sandra
authors.affiliation: Research Institute for Chromatography, President Kennedypak 26, B-8500 Kortrijk, Belgium.
authors.forename: Gerd
authors.initials: G
authors.lastname: Vanhoenacker
authors.affiliation: Research Institute for Chromatography, President Kennedypak 26, B-8500 Kortrijk, Belgium.
authors.forename: Isabel
authors.initials: I
authors.lastname: Vandenheede
authors.affiliation: Research Institute for Chromatography, President Kennedypak 26, B-8500 Kortrijk, Belgium.
authors.forename: Mieke
authors.initials: M
authors.lastname: Steenbeke
authors.affiliation: Agilent Technologies, 2850 Centerville Rd, Wilmington, DE 19808, USA.
authors.forename: Maureen
authors.initials: M
authors.lastname: Joseph
authors.affiliation: Research Institute for Chromatography, President Kennedypak 26, B-8500 Kortrijk, Belgium.
authors.forename: Pat
authors.initials: P
authors.lastname: Sandra
mesh_terms: D000080044:Ado-Trastuzumab Emtansine; D000595:Amino Acid Sequence; D061067:Antibodies, Monoclonal, Humanized / Q000737:chemistry*; D000970:Antineoplastic Agents / Q000737:chemistry*; D002853:Chromatography, Liquid / Q000295:instrumentation / Q000379:methods*; D004867:Equipment Design; D006801:Humans; D018796:Immunoconjugates / Q000737:chemistry*; D008239:Lysine / Q000737:chemistry; D008453:Maytansine / Q000031:analogs & derivatives* / Q000737:chemistry; D053719:Tandem Mass Spectrometry / Q000379:methods*; D000068878:Trastuzumab
chemical_list: D061067:Antibodies, Monoclonal, Humanized; D000970:Antineoplastic Agents; D018796:Immunoconjugates; D008453:Maytansine; D008239:Lysine; D000068878:Trastuzumab; D000080044:Ado-Trastuzumab Emtansine
keywords: Ado-trastuzumab emtansine; Antibody-drug conjugate; Comprehensive; Multiple heart-cutting; Two-dimensional liquid chromatography