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Issue 25, 2018
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Monoclonal antibody sequence assessment using a hybrid quadrupole-Orbitrap mass spectrometer

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Abstract

Primary sequence determination is a key component in the development and characterisation of biotherapeutic drug substances. Although frequently performed using peptide mapping experiments, complementary methods are necessary to confirm full sequence identification. The requirement for a multi-faceted approach to sequence verification, ensuring single amino acid variations in the primary sequence are not overlooked, is highlighted in the data presented herein. A combination of liquid chromatography-mass spectrometry (LC-MS) strategies, including bottom-up peptide mapping, middle-up mass analysis of monoclonal antibody (mAb) IdeS digested subunits and intact protein analysis, were used to assess primary sequence for anti-interleukin 8 IgG1, a recombinantly expressed mAb from Chinese hamster ovary cells. This structured approach to primary sequence evaluation provides a framework for the evaluation of mAbs and biosimilar candidates and accentuates the importance of incorporating multiple analytical methods for therapeutic protein characterisation.

Graphical abstract: Monoclonal antibody sequence assessment using a hybrid quadrupole-Orbitrap mass spectrometer

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Publication details

The article was received on 17 Mar 2018, accepted on 31 May 2018 and first published on 05 Jun 2018


Article type: Technical Note
DOI: 10.1039/C8AY00582F
Citation: Anal. Methods, 2018,10, 3100-3109
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    Monoclonal antibody sequence assessment using a hybrid quadrupole-Orbitrap mass spectrometer

    A. Farrell, S. Carillo, K. Scheffler, K. Cook and J. Bones, Anal. Methods, 2018, 10, 3100
    DOI: 10.1039/C8AY00582F

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