title: Complex mixtures of antibodies generated from a single production qualitatively and quantitatively evaluated by native Orbitrap mass spectrometry.
abstract: Composite antibody mixtures designed to combat diseases present a new, rapidly emerging technology in the field of biopharmaceuticals. The combination of multiple antibodies can lead to increased effector response and limit the effect of escape variants that can propagate the disease. However, parallel development of analytical technologies is required to provide fast, thorough, accurate, and robust characterization of these mixtures. Here, we evaluate the utility of native mass spectrometry on an Orbitrap platform with high mass resolving power to characterize composite mixtures of up to 15 separate antibodies. With this technique, unambiguous identification of each antibody in the mixtures was achieved. Mass measurements of the intact antibodies varied 7 ppm on average, allowing highly reproducible identification and quantitation of each compound in these complex mixtures. We show that with the high mass-resolving power and robustness of this technology, high-resolution native mass spectrometry can be used efficiently even for batch-to batch characterization.
authors.forename: Natalie J
authors.initials: NJ
authors.lastname: Thompson
authors.forename: Linda J A
authors.initials: LJ
authors.lastname: Hendriks
authors.forename: John
authors.initials: J
authors.lastname: de Kruif
authors.forename: Mark
authors.initials: M
authors.lastname: Throsby
authors.forename: Albert J R
authors.initials: AJ
authors.lastname: Heck
mesh_terms: D057809:HEK293 Cells; D006801:Humans; D013058:Mass Spectrometry / Q000379:methods*; D057127:Single-Chain Antibodies / Q000737:chemistry*
chemical_list: D057127:Single-Chain Antibodies