title: Characterization of monoclonal antibodies by a fast and easy liquid chromatography-mass spectrometry time-of-flight analysis on culture supernatant.
abstract: Rapid and efficient structural analysis is key to the development of new monoclonal antibodies. We have developed a fast and easy process to obtain mass spectrometry profiles of antibodies from culture supernatant. Treatment of the supernatant with IdeS generates three fragments of 25 kDa that can be analyzed by liquid chromatography-mass spectrometry time-of-flight (LC-MS TOF) in one run: LC, Fd, and Fc/2. This process gives rapid access to isoform and glycoform profiles. To specifically measure the fucosylation yield, we included a one-pot treatment with EndoS that removes the distal glycan heterogeneity. Our process was successfully compared with high-performance capillary electrophoresis with laser-induced fluorescence detection (HPCE-LIF), currently considered as the "gold standard" method.
authors: INSERM U1177 "Drugs and Molecules for Living Systems", Université de Lille, and Institut Pasteur de Lille, 59000 Lille, France.
authors: Antoine
authors: A
authors: Henninot
authors: Laboratoire Français du Fractionnement et des Biotechnologies (LFB), 91958 Courtabœuf Cedex, France.
authors: Aurelie
authors: A
authors: Terrier
authors: INSERM U1177 "Drugs and Molecules for Living Systems", Université de Lille, and Institut Pasteur de Lille, 59000 Lille, France.
authors: Julie
authors: J
authors: Charton
authors: Laboratoire Français du Fractionnement et des Biotechnologies (LFB), 91958 Courtabœuf Cedex, France.
authors: Rémi
authors: R
authors: Urbain
authors: Laboratoire Français du Fractionnement et des Biotechnologies (LFB), 91958 Courtabœuf Cedex, France.
authors: Alexandre
authors: A
authors: Fontayne
authors: INSERM U1177 "Drugs and Molecules for Living Systems", Université de Lille, and Institut Pasteur de Lille, 59000 Lille, France. Electronic address: benoit.deprez@univ-lille2.fr.
authors: Benoit
authors: B
authors: Deprez
authors: INSERM U1177 "Drugs and Molecules for Living Systems", Université de Lille, and Institut Pasteur de Lille, 59000 Lille, France.
authors: Terence
authors: T
authors: Beghyn
mesh_terms: D000911:Antibodies, Monoclonal / Q000032:analysis*; D002851:Chromatography, High Pressure Liquid; D019075:Electrophoresis, Capillary; D006031:Glycosylation; D020033:Protein Isoforms / Q000032:analysis; D013050:Spectrometry, Fluorescence; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization
chemical_list: D000911:Antibodies, Monoclonal; D020033:Protein Isoforms
keywords: Fucosylation; Glycoform; Mass spectrometry; Monoclonal antibody