Interlaboratory Study for Characterizing Monoclonal Antibodies by Top-Down and Middle-Down Mass Spectrometry
Journal of the American Society for Mass Spectrometry, vol. 31, pp. 1783–1802
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.
Authors 60
-
Affiliation as printed
Northwestern University , , ,
Northwestern University, Evanston, Illinois 60208-0001, United States
-
Affiliation as printed
Northwestern University , , ,
Northwestern University, Evanston, Illinois 60208-0001, United States
-
Yury O. Tsybin corresponding
Spectroswiss (Switzerland) · École Polytechnique Fédérale de Lausanne
Affiliation as printed
Spectroswiss , EPFL Innovation Park, Building I , ,
Spectroswiss, EPFL Innovation Park, Building I, 1015 Lausanne, Switzerland
-
University of California, Los Angeles
Affiliation as printed
University of California-Los Angeles , , ,
University of California-Los Angeles, Los Angeles, California 90095, United States
-
Affiliation as printed
Northwestern University , , ,
Northwestern University, Evanston, Illinois 60208-0001, United States
-
Affiliation as printed
Northeastern University , , ,
Northeastern University, Boston, Massachusetts 02115, United States
-
National High Magnetic Field Laboratory
Affiliation as printed
National High Magnetic Field Laboratory , , ,
National High Magnetic Field Laboratory, Tallahassee, Florida 32310, United States
-
Affiliation as printed
University of Virginia , , ,
University of Virginia, Charlottesville, Virginia 22901, United States
-
Center of Immunology Pierre Fabre
Affiliation as printed
Centre d’immunologie Pierre Fabre , ,
Centre d'immunologie Pierre Fabre, 74160 Saint-Julien-en-Genevois, France
-
The University of Texas at Austin
Affiliation as printed
University of Texas at Austin , , ,
University of Texas at Austin, Austin, Texas 78712-1224, United States
-
Yuri E. M. van der Burgt Aachen
Leiden University · Leiden University Medical Center
Affiliation as printed
Leiden University Medical Centre , ,
Leiden University Medical Centre, 2300 RC Leiden, The Netherlands
-
Affiliation as printed
Institute Pasteur , ,
Institute Pasteur, 75015 Paris, France
-
Affiliation as printed
University of Antwerp , ,
University of Antwerp, 2000 Antwerp, Belgium
-
Affiliation as printed
Biogen, Inc. , , ,
Biogen, Inc., Cambridge, Massachusetts 02142-1031, United States
-
Affiliation as printed
The University of Edinburgh , ,
The University of Edinburgh, EH9 3FJ Edinburgh, United Kingdom
-
Affiliation as printed
Consortium for Top-Down Proteomics , , ,
Consortium for Top-Down Proteomics, Cambridge, Massachusetts 02142, United States
-
Affiliation as printed
The Scripps Research Institute , , ,
The Scripps Research Institute, La Jolla, California 92037, United States
-
Affiliation as printed
University of Virginia , , ,
University of Virginia, Charlottesville, Virginia 22901, United States
-
Affiliation as printed
Institute Pasteur , ,
Institute Pasteur, 75015 Paris, France
-
École Polytechnique Fédérale de Lausanne
Affiliation as printed
Ecole Polytechnique Fédérale de Lausanne , ,
Ecole Polytechnique Fédérale de Lausanne, 1015 Lausanne, Switzerland
-
University of Wisconsin–Madison
Affiliation as printed
University of Wisconsin-Madison , , ,
University of Wisconsin-Madison, Madison, Wisconsin 53706, United States
-
University of Maryland, Baltimore · University of Maryland, College Park
Affiliation as printed
University of Maryland , , ,
University of Maryland, Baltimore, Maryland 21201, United States
-
University of Maryland, Baltimore · University of Maryland, College Park
Affiliation as printed
University of Maryland , , ,
University of Maryland, Baltimore, Maryland 21201, United States
-
The University of Texas at Austin
Affiliation as printed
University of Texas at Austin , , ,
University of Texas at Austin, Austin, Texas 78712-1224, United States
-
Affiliation as printed
Novo Nordisk , ,
Novo Nordisk, DK-2760 Malov, Denmark
-
National High Magnetic Field Laboratory
Affiliation as printed
National High Magnetic Field Laboratory , , ,
National High Magnetic Field Laboratory, Tallahassee, Florida 32310, United States
-
National High Magnetic Field Laboratory
Affiliation as printed
National High Magnetic Field Laboratory , , ,
National High Magnetic Field Laboratory, Tallahassee, Florida 32310, United States
-
Affiliation as printed
University of Virginia , , ,
University of Virginia, Charlottesville, Virginia 22901, United States
-
Affiliation as printed
Baylor College of Medicine , , ,
Baylor College of Medicine, Houston, Texas 77030-3411, United States
-
Affiliation as printed
The University of Edinburgh , ,
The University of Edinburgh, EH9 3FJ Edinburgh, United Kingdom
-
Affiliation as printed
University of Virginia , , ,
University of Virginia, Charlottesville, Virginia 22901, United States
-
Affiliation as printed
Northwestern University , , ,
Northwestern University, Evanston, Illinois 60208-0001, United States
-
Spectroswiss (Switzerland) · École Polytechnique Fédérale de Lausanne
Affiliation as printed
Spectroswiss , EPFL Innovation Park, Building I , ,
Spectroswiss, EPFL Innovation Park, Building I, 1015 Lausanne, Switzerland
-
University of Wisconsin–Madison
Affiliation as printed
University of Wisconsin-Madison , , ,
University of Wisconsin-Madison, Madison, Wisconsin 53706, United States
-
Affiliation as printed
Institute Pasteur , ,
Institute Pasteur, 75015 Paris, France
-
Florida State University · National High Magnetic Field Laboratory
Affiliation as printed
Florida State University , , ,
National High Magnetic Field Laboratory , , ,
Florida State University, Tallahassee, Florida 32310-4005, United States
National High Magnetic Field Laboratory, Tallahassee, Florida 32310, United States
-
École Polytechnique Fédérale de Lausanne
Affiliation as printed
Ecole Polytechnique Fédérale de Lausanne , ,
Ecole Polytechnique Fédérale de Lausanne, 1015 Lausanne, Switzerland
-
University of Wisconsin–Madison
Affiliation as printed
University of Wisconsin-Madison , , ,
University of Wisconsin-Madison, Madison, Wisconsin 53706, United States
-
Spectroswiss (Switzerland) · École Polytechnique Fédérale de Lausanne
Affiliation as printed
Spectroswiss , EPFL Innovation Park, Building I , ,
Spectroswiss, EPFL Innovation Park, Building I, 1015 Lausanne, Switzerland
-
Simone Nicolardi Aachen
Leiden University · Leiden University Medical Center
Affiliation as printed
Leiden University Medical Centre , ,
Leiden University Medical Centre, 2300 RC Leiden, The Netherlands
-
Pacific Northwest National Laboratory
Affiliation as printed
Pacific Northwest National Laboratory , , ,
Pacific Northwest National Laboratory, Richland, Washington 99354, United States
-
Affiliation as printed
Bruker Daltonik GmbH , ,
Bruker Daltonik GmbH, 28359 Bremen, Germany
-
University of California, Los Angeles
Affiliation as printed
University of California-Los Angeles , , ,
University of California-Los Angeles, Los Angeles, California 90095, United States
-
Affiliation as printed
Bruker Daltonik GmbH , ,
Bruker Daltonik GmbH, 28359 Bremen, Germany
-
Affiliation as printed
Genentech, Inc. , , ,
Genentech, Inc., South San Francisco, California 94080-4990, United States
-
Affiliation as printed
Biogen, Inc. , , ,
Biogen, Inc., Cambridge, Massachusetts 02142-1031, United States
-
Affiliation as printed
Northeastern University , , ,
Northeastern University, Boston, Massachusetts 02115, United States
-
Affiliation as printed
University of Virginia , , ,
University of Virginia, Charlottesville, Virginia 22901, United States
-
Pacific Northwest National Laboratory
Affiliation as printed
Pacific Northwest National Laboratory , , ,
Pacific Northwest National Laboratory, Richland, Washington 99354, United States
-
University of Wisconsin–Madison
Affiliation as printed
University of Wisconsin-Madison , , ,
University of Wisconsin-Madison, Madison, Wisconsin 53706, United States
-
University of Wisconsin–Madison
Affiliation as printed
University of Wisconsin-Madison , , ,
University of Wisconsin-Madison, Madison, Wisconsin 53706, United States
-
University of Leeds · University of Antwerp
Affiliation as printed
University of Antwerp , ,
University of Leeds , ,
University of Antwerp, 2000 Antwerp, Belgium
University of Leeds, LS2 9JT Leeds, United Kingdom
-
Affiliation as printed
Bruker Daltonik GmbH , ,
Bruker Daltonik GmbH, 28359 Bremen, Germany
-
Affiliation as printed
Northwestern University , , ,
Northwestern University, Evanston, Illinois 60208-0001, United States
-
National High Magnetic Field Laboratory
Affiliation as printed
National High Magnetic Field Laboratory , , ,
National High Magnetic Field Laboratory, Tallahassee, Florida 32310, United States
-
Washington University in St. Louis
Affiliation as printed
Washington University School of Medicine , , ,
Washington University School of Medicine, St. Louis, Missouri 63110, United States
-
Affiliation as printed
The Scripps Research Institute , , ,
The Scripps Research Institute, La Jolla, California 92037, United States
-
University of Maryland, Baltimore · University of Maryland, College Park
Affiliation as printed
University of Maryland , , ,
University of Maryland, Baltimore, Maryland 21201, United States
-
Affiliation as printed
Baylor College of Medicine , , ,
Baylor College of Medicine, Houston, Texas 77030-3411, United States
-
Pacific Northwest National Laboratory
Affiliation as printed
Pacific Northwest National Laboratory , , ,
Pacific Northwest National Laboratory, Richland, Washington 99354, United States
Cited by 113 stored of 130
No patents citing this paper on Lens.org (checked 2026-10-11).