@article{811223ace4454699aa953fc42638a2c4,
title = "Interlaboratory Study for Characterizing Monoclonal Antibodies by Top-Down and Middle-Down Mass Spectrometry",
abstract = "The Consortium for Top-Down Proteomics (www.topdownproteomics.org) launched the present study to assess the current state of top-down mass spectrometry (TD MS) and middle-down mass spectrometry (MD MS) for characterizing monoclonal antibody (mAb) primary structures, including their modifications. To meet the needs of the rapidly growing therapeutic antibody market, it is important to develop analytical strategies to characterize the heterogeneity of a therapeutic product's primary structure accurately and reproducibly. The major objective of the present study is to determine whether current TD/MD MS technologies and protocols can add value to the more commonly employed bottom-up (BU) approaches with regard to confirming protein integrity, sequencing variable domains, avoiding artifacts, and revealing modifications and their locations. We also aim to gather information on the common TD/MD MS methods and practices in the field. A panel of three mAbs was selected and centrally provided to 20 laboratories worldwide for the analysis: Sigma mAb standard (SiLuLite), NIST mAb standard, and the therapeutic mAb Herceptin (trastuzumab). Various MS instrument platforms and ion dissociation techniques were employed. The present study confirms that TD/MD MS tools are available in laboratories worldwide and provide complementary information to the BU approach that can be crucial for comprehensive mAb characterization. The current limitations, as well as possible solutions to overcome them, are also outlined. A primary limitation revealed by the results of the present study is that the expert knowledge in both experiment and data analysis is indispensable to practice TD/MD MS.",
keywords = "FTMS, Fourier transform mass spectrometry, MS/MS, Therapeutic protein, glycoform, intact mass measurement, tandem mass spectrometry",
author = "Kristina Srzenti{\'c} and Luca Fornelli and Tsybin, {Yury O.} and Loo, {Joseph A.} and Henrique Seckler and Agar, {Jeffrey N.} and Anderson, {Lissa C.} and Bai, {Dina L.} and Alain Beck and Brodbelt, {Jennifer S.} and {van der Burgt}, {Yuri E.M.} and Julia Chamot-Rooke and Sneha Chatterjee and Yunqiu Chen and Clarke, {David J.} and Danis, {Paul O.} and Diedrich, {Jolene K.} and D'Ippolito, {Robert A.} and Mathieu Dupr{\'e} and Natalia Gasilova and Ying Ge and Goo, {Young Ah} and Goodlett, {David R.} and Sylvester Greer and Haselmann, {Kim F.} and Lidong He and Hendrickson, {Christopher L.} and Hinkle, {Joshua D.} and Holt, {Matthew V.} and Sam Hughes and Hunt, {Donald F.} and Kelleher, {Neil L.} and Kozhinov, {Anton N.} and Ziqing Lin and Christian Malosse and Marshall, {Alan G.} and Laure Menin and Millikin, {Robert J.} and Nagornov, {Konstantin O.} and Simone Nicolardi and Ljiljana Pa{\v s}a-Toli{\'c} and Stuart Pengelley and Quebbemann, {Neil R.} and Anja Resemann and Wendy Sandoval and Richa Sarin and Schmitt, {Nicholas D.} and Jeffrey Shabanowitz and Shaw, {Jared B.} and Shortreed, {Michael R.} and Smith, {Lloyd M.} and Frank Sobott and Detlev Suckau and Timothy Toby and Weisbrod, {Chad R.} and Wildburger, {Norelle C.} and Yates, {John R.} and Yoon, {Sung Hwan} and Young, {Nicolas L.} and Mowei Zhou",
note = "Funding Information: We greatly appreciate the support of the Pilot Project by the Consortium for Top-Down Proteomics and its supporting organizations, Thermo Fisher Scientific, Inc., Bruker Corp., and Genovis AB. We also acknowledge Genovis for providing enzymes (IdeS and KGP) and Millipore Sigma for donation of their mAb standard (SiLuLite mAb). J.A.L. acknowledges support from the US National Institutes of Health (R01GM103479, S10RR028893, S10 OD018504); Y.O.T. and J.C.R. from European Horizon 2020 research and innovation program under Grant Agreement No. 829157 (TopSpec); J.S.B. from the Welch Foundation (F-1155) and the National Science Foundation (CHE1402753); YG from NIH GM125085 and S10OD018475; and D.F.H. from the NIH Grant GM037537. L.P.T., J.B.S., and M.Z. acknowledge the support from the Intramural program at Environmental Molecular Sciences Laboratory (EMSL), a DOE Office of Science User Facility sponsored by the Office of Biological and Environmental Research and operated under Contract No. DE-AC05-76RL01830. C.L.H., L.C.A., L.H., and A.G.M. acknowledge support for work performed at the National High Magnetic Field Laboratory, which is supported by National Science Foundation Cooperative Agreement No. DMR-1644779 and the State of Florida. This research was carried out in collaboration with the National Resource for Translational and Developmental Proteomics under Grant P41 GM108569 from the National Institute of General Medical Sciences, National Institutes of Health. Funding Information: We greatly appreciate the support of the Pilot Project by the Consortium for Top-Down Proteomics and its supporting organizations Thermo Fisher Scientific, Inc., Bruker Corp., and Genovis AB. We also acknowledge Genovis for providing enzymes (IdeS and KGP) and Millipore Sigma for donation of their mAb standard (SiLuLite mAb). J.A.L. acknowledges support from the US National Institutes of Health (R01GM103479, S10RR028893 S10 OD018504); Y.O.T. and J.C.R. from European Horizon 2020 research and innovation program under Grant Agreement No. 829157 (TopSpec); J.S.B. from the Welch Foundation (F-1155) and the National Science Foundation (CHE1402753); YG from NIH GM125085 and S10OD018475; and D.F.H. from the NIH Grant GM037537. L.P.T., J.B.S., and M.Z. acknowledge the support from the Intramural program at Environmental Molecular Sciences Laboratory (EMSL), a DOE Office of Science User Facility sponsored by the Office of Biological and Environmental Research and operated under Contract No. DE-AC05-76RL01830. C.L.H., L.C.A., L.H. and A.G.M. acknowledge support for work performed at the National High Magnetic Field Laboratory, which is supported by National Science Foundation Cooperative Agreement No. DMR-1644779 and the State of Florida. This research was carried out in collaboration with the National Resource for Translational and Developmental Proteomics under Grant P41 GM108569 from the National Institute of General Medical Sciences National Institutes of Health. Publisher Copyright: {\textcopyright} 2020 Springer New York LLC. All rights reserved.",
year = "2020",
month = sep,
day = "2",
doi = "10.1021/jasms.0c00036",
language = "English (US)",
volume = "31",
pages = "1783--1802",
journal = "Journal of the American Society for Mass Spectrometry",
issn = "1044-0305",
publisher = "Springer New York",
number = "9",
}