Structural analysis of monoclonal antibodies by ultrahigh resolution MALDI in-source decay FT-ICR mass spectrometry

Van der Burgt, YEM, Kilgour, DPA ORCID: https://orcid.org/0000-0002-3860-7532, Tsybin, YO, Srzentić, K, Fornelli, L, Beck, A, Wuhrer, M and Nicolardi, S, 2019. Structural analysis of monoclonal antibodies by ultrahigh resolution MALDI in-source decay FT-ICR mass spectrometry. Analytical Chemistry, 91 (3), pp. 2079-2085. ISSN 0003-2700

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Abstract

The emergence of complex protein therapeutics in general and monoclonal antibodies (mAbs) in particular have stimulated analytical chemists to develop new methods and strategies for their structural characterization. Mass spectrometry plays a key role in providing information on the primary amino acid sequence, post-translational modifications, and other structure characteristics that must be monitored during the manufacturing process and subsequent quality control assessment. In this study, we present a novel method that allows structural characterization of mAbs based on MALDI in-source decay (ISD) fragmentation, coupled with Fourier transform ion cyclotron resonance (FT-ICR) MS. The method benefits from higher resolution of absorption mode FT mass spectra, compared to magnitude mode, which enables simultaneous identification of ISD fragments from both the heavy and light chains with a higher confidence in a wide mass range up to m/z 13 500. This method was applied to two standard mAbs, namely NIST mAb and trastuzumab, in preparation for method application in an interlaboratory study on mAbs structural analysis coordinated by the Consortium for Top-Down Proteomics. Extensive sequence coverage was obtained from the middle-down analysis (IdeS- and GingisKHAN-digested mAbs) that complemented the top-down analysis of intact mAbs. In addition, MALDI FT-ICR MS of IdeS-digested mAbs allowed isotopic-level profiling of proteoforms with regard to heavy chain N-glycosylation.

Item Type:	Journal article
Publication Title:	Analytical Chemistry
Creators:	Van der Burgt, Y.E.M., Kilgour, D.P.A., Tsybin, Y.O., Srzentić, K., Fornelli, L., Beck, A., Wuhrer, M. and Nicolardi, S.
Publisher:	American Chemical Society
Date:	2019
Volume:	91
Number:	3
ISSN:	0003-2700
Identifiers:	Number Type 10.1021/acs.analchem.8b04515 DOI 689623 Other
Rights:	This is an open access article published under a Creative Commons Non-Commercial No Derivative Works (CC-BY-NC-ND) Attribution License, which permits copying and redistribution of the article, and creation of adaptations, all for non-commercial purposes.
Divisions:	Schools > School of Science and Technology
Record created by:	Jonathan Gallacher
Date Added:	10 Jan 2019 09:33
Last Modified:	10 Oct 2019 11:02
URI:	https://irep.ntu.ac.uk/id/eprint/35520

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