skip to main content
DOE PAGES title logo U.S. Department of Energy
Office of Scientific and Technical Information

Title: Enabling adoption of 2D-NMR for the higher order structure assessment of monoclonal antibody therapeutics

Abstract

The increased interest in using monoclonal antibodies (mAbs) as a platform for biopharmaceuticals has led to the need for new analytical techniques that can precisely assess physicochemical properties of these large and very complex drugs for the purpose of correctly identifying quality attributes (QA). One QA, higher order structure (HOS), is unique to biopharmaceuticals and essential for establishing consistency in biopharmaceutical manufacturing, detecting process-related variations from manufacturing changes and establishing comparability between biologic products. To address this measurement challenge, two-dimensional nuclear magnetic resonance spectroscopy (2D-NMR) methods were introduced that allow for the precise atomic-level comparison of the HOS between two proteins, including mAbs. Here, an inter-laboratory comparison involving 26 industrial, government and academic laboratories worldwide was performed as a benchmark using the NISTmAb, from the National Institute of Standards and Technology (NIST), to facilitate the translation of the 2D-NMR method into routine use for biopharmaceutical product development. Two-dimensional 1H,15N and 1H,13C NMR spectra were acquired with harmonized experimental protocols on the unlabeled Fab domain and a uniformly enriched-15N, 20%-13C-enriched system suitability sample derived from the NISTmAb. Chemometric analyses from over 400 spectral maps acquired on 39 different NMR spectrometers ranging from 500 MHz to 900 MHz demonstrate spectral fingerprints thatmore » are fit-for-purpose for the assessment of HOS. The 2D-NMR method is shown to provide the measurement reliability needed to move the technique from an emerging technology to a harmonized, routine measurement that can be generally applied with great confidence to high precision assessments of the HOS of mAb-based biotherapeutics.« less

Authors:
 [1];  [1];  [1]; ORCiD logo [1];  [2];  [2];  [3]; ORCiD logo [3]; ORCiD logo [3];  [4]; ORCiD logo [4]; ORCiD logo [4];  [5];  [5];  [6]; ORCiD logo [7];  [7];  [8];  [9]; ORCiD logo [9] more »; ORCiD logo [9];  [10];  [10];  [11];  [11]; ORCiD logo [12];  [13]; ORCiD logo [14]; ORCiD logo [14]; ORCiD logo [15];  [15];  [16];  [17]; ORCiD logo [18]; ORCiD logo [18]; ORCiD logo [18];  [19];  [19];  [20]; ORCiD logo [20];  [21];  [22];  [23];  [24];  [25]; ORCiD logo [26]; ORCiD logo [26];  [27];  [27] « less
+ Show Author Affiliations
  1. National Institute of Standards and Technology and the University of Maryland, Rockville, MD (United States)
  2. National Inst. of Standards and Technology (NIST), Gaithersburg, MD (United States)
  3. Centre for Biologics Evaluation, Ottawa, ON (Canada)
  4. Univ. of Queensland, St. Lucia, QLD (Australia)
  5. Univ. of Melbourne (Australia)
  6. Univ. of Bayreuth (Germany)
  7. Stockholm Univ. (Sweden)
  8. Analytical R&D, Pfizer Essential Health, Lake Forest, IL (United States)
  9. Pacific Northwest National Lab. (PNNL), Richland, WA (United States)
  10. National Research Council of Canada, Montreal, Quebec (Canada)
  11. National Institutes of Natural Sciences, Myodaiji, Okazaki (Japan)
  12. Univ. of South Carolina, Charleston, SC (United States)
  13. Momenta Pharmaceuticals, Cambridge, MA (United States)
  14. Swedish Medical Products Agency, Laboratory, Uppsala (Sweden)
  15. Leibniz-Forschungsinstitut für Molekulare Pharmakologie (FMP), Berlin (Germany)
  16. Roche Innovation Center Basel (Switzerland)
  17. Genentech, South San Francisco, CA (United States)
  18. Center for Drug Evaluation and Research, Food and Drug Administration, MD (United States)
  19. Institute of Molecular Biology and Biophysics, ETH Zurich (Switzerland)
  20. NMR Centre, Ljubljana (Slovenia)
  21. Eli Lilly and Company, Indianapolis, IN (United States)
  22. Bruker BioSpin Corporation, Billerica, MA (United States)
  23. Brazilian Biosciences National Laboratory (LNBio), Brazilian Center for Research in Energy and Materials (CNPEM), Campinas (Brazil)
  24. Brazilian Synchrotron Light Laboratory (LNLS), Brazilian Center for Research in Energy and Materials (CNPEM), Campinas (Brazil)
  25. Biogen Inc., Cambridge, MA (United States)
  26. Baxter Pharmaceuticals R&D, Baxter Healthcare, Round Lake, IL (United States)
  27. Amgen Inc., Thousand Oaks, CA (United States)
Publication Date:
Research Org.:
Pacific Northwest National Lab. (PNNL), Richland, WA (United States)
Sponsoring Org.:
USDOE
OSTI Identifier:
1545065
Report Number(s):
PNNL-SA-131540
Journal ID: ISSN 1942-0862
Grant/Contract Number:  
AC05-76RL01830
Resource Type:
Accepted Manuscript
Journal Name:
mAbs
Additional Journal Information:
Journal Volume: 11; Journal Issue: 1; Journal ID: ISSN 1942-0862
Publisher:
Taylor & Francis
Country of Publication:
United States
Language:
English
Subject:
59 BASIC BIOLOGICAL SCIENCES; 37 INORGANIC, ORGANIC, PHYSICAL, AND ANALYTICAL CHEMISTRY; monoclonal antibody (mAb) therapeutics; NISTmAb; higher order structure; nuclear magnetic resonance spectroscopy (NMR); comparability; chemometrics

Citation Formats

Brinson, Robert G., Marino, John P., Delaglio, Frank, Arbogast, Luke W., Evans, Ryan M., Kearsley, Anthony, Gingras, Geneviève, Ghasriani, Houman, Aubin, Yves, Pierens, Gregory K., Jia, Xinying, Mobli, Mehdi, Grant, Hamish G., Keizer, David W., Schweimer, Kristian, Ståhle, Jonas, Widmalm, Göran, Zartler, Edward R., Lawrence, Chad W., Reardon, Patrick N., Cort, John R., Xu, Ping, Ni, Feng, Yanaka, Saeko, Kato, Koichi, Parnham, Stuart R., Tsao, Desiree, Blomgren, Andreas, Rundlöf, Torgny, Trieloff, Nils, Schmieder, Peter, Ross, Alfred, Skidmore, Ken, Chen, Kang, Keire, David, Freedberg, Darón I., Suter-Stahel, Thea, Wider, Gerhard, Ilc, Gregor, Plavec, Janez, Bradley, Scott A., Baldisseri, Donna M., Sforça, Mauricio Luis, Zeri, Ana Carolina de Mattos, Wei, Julie Yu, Szabo, Christina M., Amezcua, Carlos A., Jordan, John B., and Wikström, Mats. Enabling adoption of 2D-NMR for the higher order structure assessment of monoclonal antibody therapeutics. United States: N. p., 2018. Web. doi:10.1080/19420862.2018.1544454.
Copy to clipboard
Brinson, Robert G., Marino, John P., Delaglio, Frank, Arbogast, Luke W., Evans, Ryan M., Kearsley, Anthony, Gingras, Geneviève, Ghasriani, Houman, Aubin, Yves, Pierens, Gregory K., Jia, Xinying, Mobli, Mehdi, Grant, Hamish G., Keizer, David W., Schweimer, Kristian, Ståhle, Jonas, Widmalm, Göran, Zartler, Edward R., Lawrence, Chad W., Reardon, Patrick N., Cort, John R., Xu, Ping, Ni, Feng, Yanaka, Saeko, Kato, Koichi, Parnham, Stuart R., Tsao, Desiree, Blomgren, Andreas, Rundlöf, Torgny, Trieloff, Nils, Schmieder, Peter, Ross, Alfred, Skidmore, Ken, Chen, Kang, Keire, David, Freedberg, Darón I., Suter-Stahel, Thea, Wider, Gerhard, Ilc, Gregor, Plavec, Janez, Bradley, Scott A., Baldisseri, Donna M., Sforça, Mauricio Luis, Zeri, Ana Carolina de Mattos, Wei, Julie Yu, Szabo, Christina M., Amezcua, Carlos A., Jordan, John B., & Wikström, Mats. Enabling adoption of 2D-NMR for the higher order structure assessment of monoclonal antibody therapeutics. United States. doi:10.1080/19420862.2018.1544454.
Copy to clipboard
Brinson, Robert G., Marino, John P., Delaglio, Frank, Arbogast, Luke W., Evans, Ryan M., Kearsley, Anthony, Gingras, Geneviève, Ghasriani, Houman, Aubin, Yves, Pierens, Gregory K., Jia, Xinying, Mobli, Mehdi, Grant, Hamish G., Keizer, David W., Schweimer, Kristian, Ståhle, Jonas, Widmalm, Göran, Zartler, Edward R., Lawrence, Chad W., Reardon, Patrick N., Cort, John R., Xu, Ping, Ni, Feng, Yanaka, Saeko, Kato, Koichi, Parnham, Stuart R., Tsao, Desiree, Blomgren, Andreas, Rundlöf, Torgny, Trieloff, Nils, Schmieder, Peter, Ross, Alfred, Skidmore, Ken, Chen, Kang, Keire, David, Freedberg, Darón I., Suter-Stahel, Thea, Wider, Gerhard, Ilc, Gregor, Plavec, Janez, Bradley, Scott A., Baldisseri, Donna M., Sforça, Mauricio Luis, Zeri, Ana Carolina de Mattos, Wei, Julie Yu, Szabo, Christina M., Amezcua, Carlos A., Jordan, John B., and Wikström, Mats. Thu . "Enabling adoption of 2D-NMR for the higher order structure assessment of monoclonal antibody therapeutics". United States. doi:10.1080/19420862.2018.1544454. https://www.osti.gov/servlets/purl/1545065.
Copy to clipboard
@article{osti_1545065,
title = {Enabling adoption of 2D-NMR for the higher order structure assessment of monoclonal antibody therapeutics},
author = {Brinson, Robert G. and Marino, John P. and Delaglio, Frank and Arbogast, Luke W. and Evans, Ryan M. and Kearsley, Anthony and Gingras, Geneviève and Ghasriani, Houman and Aubin, Yves and Pierens, Gregory K. and Jia, Xinying and Mobli, Mehdi and Grant, Hamish G. and Keizer, David W. and Schweimer, Kristian and Ståhle, Jonas and Widmalm, Göran and Zartler, Edward R. and Lawrence, Chad W. and Reardon, Patrick N. and Cort, John R. and Xu, Ping and Ni, Feng and Yanaka, Saeko and Kato, Koichi and Parnham, Stuart R. and Tsao, Desiree and Blomgren, Andreas and Rundlöf, Torgny and Trieloff, Nils and Schmieder, Peter and Ross, Alfred and Skidmore, Ken and Chen, Kang and Keire, David and Freedberg, Darón I. and Suter-Stahel, Thea and Wider, Gerhard and Ilc, Gregor and Plavec, Janez and Bradley, Scott A. and Baldisseri, Donna M. and Sforça, Mauricio Luis and Zeri, Ana Carolina de Mattos and Wei, Julie Yu and Szabo, Christina M. and Amezcua, Carlos A. and Jordan, John B. and Wikström, Mats},
abstractNote = {The increased interest in using monoclonal antibodies (mAbs) as a platform for biopharmaceuticals has led to the need for new analytical techniques that can precisely assess physicochemical properties of these large and very complex drugs for the purpose of correctly identifying quality attributes (QA). One QA, higher order structure (HOS), is unique to biopharmaceuticals and essential for establishing consistency in biopharmaceutical manufacturing, detecting process-related variations from manufacturing changes and establishing comparability between biologic products. To address this measurement challenge, two-dimensional nuclear magnetic resonance spectroscopy (2D-NMR) methods were introduced that allow for the precise atomic-level comparison of the HOS between two proteins, including mAbs. Here, an inter-laboratory comparison involving 26 industrial, government and academic laboratories worldwide was performed as a benchmark using the NISTmAb, from the National Institute of Standards and Technology (NIST), to facilitate the translation of the 2D-NMR method into routine use for biopharmaceutical product development. Two-dimensional 1H,15N and 1H,13C NMR spectra were acquired with harmonized experimental protocols on the unlabeled Fab domain and a uniformly enriched-15N, 20%-13C-enriched system suitability sample derived from the NISTmAb. Chemometric analyses from over 400 spectral maps acquired on 39 different NMR spectrometers ranging from 500 MHz to 900 MHz demonstrate spectral fingerprints that are fit-for-purpose for the assessment of HOS. The 2D-NMR method is shown to provide the measurement reliability needed to move the technique from an emerging technology to a harmonized, routine measurement that can be generally applied with great confidence to high precision assessments of the HOS of mAb-based biotherapeutics.},
doi = {10.1080/19420862.2018.1544454},
journal = {mAbs},
number = 1,
volume = 11,
place = {United States},
year = {2018},
month = {12}
}
Copy to clipboard
Journal Article:
Free Publicly Available Full Text
Publisher's Version of Record
DOI:  10.1080/19420862.2018.1544454
Copyright Statement
Other availability
Search WorldCat Search WorldCat to find libraries that may hold this journal
Citation Metrics:
Cited by: 5 works
Citation information provided by
Web of Science
Save / Share:
Export Metadata
Save to My Library
You must Sign In or Create an Account in order to save documents to your library.

Works referenced in this record:

Technical Decision Making With Higher Order Structure Data: Perspectives on Higher Order Structure Characterization From the Biopharmaceutical Industry
journal, December 2016

  • Weiss, William F.; Gabrielson, John P.; Al-Azzam, Wasfi
  • Journal of Pharmaceutical Sciences, Vol. 105, Issue 12
  • DOI: 10.1016/j.xphs.2016.09.003

Application of iterative soft thresholding for fast reconstruction of NMR data non-uniformly sampled with multidimensional Poisson Gap scheduling
journal, February 2012

  • Hyberts, Sven G.; Milbradt, Alexander G.; Wagner, Andreas B.
  • Journal of Biomolecular NMR, Vol. 52, Issue 4
  • DOI: 10.1007/s10858-012-9611-z

What NMR can do in the biopharmaceutical industry
journal, January 2018

  • Kiss, Róbert; Fizil, Ádám; Szántay, Csaba
  • Journal of Pharmaceutical and Biomedical Analysis, Vol. 147
  • DOI: 10.1016/j.jpba.2017.07.004

NMRFAM-SPARKY: enhanced software for biomolecular NMR spectroscopy
journal, December 2014

Precision and robustness of 2D-NMR for structure assessment of filgrastim biosimilars
journal, February 2016

  • Ghasriani, Houman; Hodgson, Derek J.; Brinson, Robert G.
  • Nature Biotechnology, Vol. 34, Issue 2
  • DOI: 10.1038/nbt.3474

The importance of not saturating water in protein NMR. Application to sensitivity enhancement and NOE measurements
journal, December 1993

  • Grzesiek, Stephan; Bax, Ad
  • Journal of the American Chemical Society, Vol. 115, Issue 26
  • DOI: 10.1021/ja00079a052

Gradient-Tailored Water Suppression for 1H-15N HSQC Experiments Optimized to Retain Full Sensitivity
journal, April 1993

  • Sklenar, V.; Piotto, M.; Leppik, R.
  • Journal of Magnetic Resonance, Series A, Vol. 102, Issue 2
  • DOI: 10.1006/jmra.1993.1098

Multivariate Analysis of Two-Dimensional 1 H, 13 C Methyl NMR Spectra of Monoclonal Antibody Therapeutics To Facilitate Assessment of Higher Order Structure
journal, October 2017

Biosimilar structural comparability assessment by NMR: from small proteins to monoclonal antibodies
journal, August 2016

  • Japelj, Boštjan; Ilc, Gregor; Marušič, Jaka
  • Scientific Reports, Vol. 6, Issue 1
  • DOI: 10.1038/srep32201

Antibodies to watch in 2018
journal, January 2018

Nonuniform sampling and non-Fourier signal processing methods in multidimensional NMR
journal, November 2014

Enhanced biosynthetically directed fractional carbon-13 enrichment of proteins for backbone NMR assignments
journal, November 2015

  • Wenrich, Broc R.; Sonstrom, Reilly E.; Gupta, Riju A.
  • Protein Expression and Purification, Vol. 115
  • DOI: 10.1016/j.pep.2015.08.005

Deterministic multidimensional nonuniform gap sampling
journal, December 2015

Analytical tools for characterizing biopharmaceuticals and the implications for biosimilars
journal, June 2012

  • Berkowitz, Steven A.; Engen, John R.; Mazzeo, Jeffrey R.
  • Nature Reviews Drug Discovery, Vol. 11, Issue 7
  • DOI: 10.1038/nrd3746

Advancing pharmaceutical quality: An overview of science and research in the U.S. FDA’s Office of Pharmaceutical Quality
journal, December 2016

Technical Decision-Making with Higher Order Structure Data: Starting a New Dialogue
journal, April 2015

  • Gabrielson, John P.; Weiss, William F.
  • Journal of Pharmaceutical Sciences, Vol. 104, Issue 4
  • DOI: 10.1002/jps.24393

Poisson-Gap Sampling and Forward Maximum Entropy Reconstruction for Enhancing the Resolution and Sensitivity of Protein NMR Data
journal, February 2010

  • Hyberts, Sven G.; Takeuchi, Koh; Wagner, Gerhard
  • Journal of the American Chemical Society, Vol. 132, Issue 7
  • DOI: 10.1021/ja908004w

Effect of Polysorbate 20 and Polysorbate 80 on the Higher-Order Structure of a Monoclonal Antibody and Its Fab and Fc Fragments Probed Using 2D Nuclear Magnetic Resonance Spectroscopy
journal, December 2017

  • Singh, Surinder M.; Bandi, Swati; Jones, David N. M.
  • Journal of Pharmaceutical Sciences, Vol. 106, Issue 12
  • DOI: 10.1016/j.xphs.2017.08.011

SOFAST-HMQC Experiments for Recording Two-dimensional Deteronuclear Correlation Spectra of Proteins within a Few Seconds
journal, December 2005

  • Schanda, Paul; Kupče, Ēriks; Brutscher, Bernhard
  • Journal of Biomolecular NMR, Vol. 33, Issue 4
  • DOI: 10.1007/s10858-005-4425-x

NMRPipe: A multidimensional spectral processing system based on UNIX pipes
journal, November 1995

  • Delaglio, Frank; Grzesiek, Stephan; Vuister, GeertenW.
  • Journal of Biomolecular NMR, Vol. 6, Issue 3
  • DOI: 10.1007/BF00197809

Monitoring Effects of Excipients, Formulation Parameters and Mutations on the High Order Structure of Filgrastim by NMR
journal, June 2015

  • Aubin, Yves; Hodgson, Derek J.; Thach, William B.
  • Pharmaceutical Research, Vol. 32, Issue 10
  • DOI: 10.1007/s11095-015-1713-3

Mapping Monoclonal Antibody Structure by 2D 13 C NMR at Natural Abundance
journal, March 2015

  • Arbogast, Luke W.; Brinson, Robert G.; Marino, John P.
  • Analytical Chemistry, Vol. 87, Issue 7
  • DOI: 10.1021/ac504804m

The NISTmAb Reference Material 8671 value assignment, homogeneity, and stability
journal, February 2018

  • Schiel, John E.; Turner, Abby; Mouchahoir, Trina
  • Analytical and Bioanalytical Chemistry, Vol. 410, Issue 8
  • DOI: 10.1007/s00216-017-0800-1

The NISTmAb Reference Material 8671 lifecycle management and quality plan
journal, February 2018

Assessment of the higher order structure of Humira®, Remicade®, Avastin®, Rituxan®, Herceptin®, and Enbrel® by 2D-NMR fingerprinting
journal, January 2019

  • Hodgson, Derek J.; Ghasriani, Houman; Aubin, Yves
  • Journal of Pharmaceutical and Biomedical Analysis, Vol. 163
  • DOI: 10.1016/j.jpba.2018.09.056

2D 1HN, 15N Correlated NMR Methods at Natural Abundance for Obtaining Structural Maps and Statistical Comparability of Monoclonal Antibodies
journal, October 2015

  • Arbogast, Luke W.; Brinson, Robert G.; Formolo, Trina
  • Pharmaceutical Research, Vol. 33, Issue 2
  • DOI: 10.1007/s11095-015-1802-3

Using chemical shift perturbation to characterise ligand binding
journal, August 2013

    [ × clear filter / sort ]

    Works referencing / citing this record:

      [ × clear filter / sort ]

      Similar records in OSTI.GOV collections: