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

Robert G Brinson; John P Marino; Frank Delaglio; Luke W Arbogast; Ryan M Evans; Anthony Kearsley; Geneviève Gingras; Houman Ghasriani; Yves Aubin; Gregory K Pierens; Xinying Jia; Mehdi Mobli; Hamish G Grant; David W Keizer; Kristian Schweimer; Jonas Ståhle; Göran Widmalm; Edward R Zartler; Chad W Lawrence; Patrick N Reardon; John R Cort; Ping Xu; Feng Ni; Saeko Yanaka; Koichi Kato; Stuart R Parnham; Desiree Tsao; Andreas Blomgren; Torgny Rundlöf; Nils Trieloff; Peter Schmieder; Alfred Ross; Ken Skidmore; Kang Chen; David Keire; Darón I Freedberg; Thea Suter-Stahel; Gerhard Wider; Gregor Ilc; Janez Plavec; Scott A Bradley; Donna M Baldisseri; Mauricio Luis Sforça; Ana Carolina de Mattos Zeri; Julie Yu Wei; Christina M Szabo; Carlos A Amezcua; John B Jordan; Mats Wikström

doi:10.1080/19420862.2018.1544454

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

MAbs. 2019 Jan;11(1):94-105. doi: 10.1080/19420862.2018.1544454. Epub 2018 Dec 22.

Authors

Robert G Brinson¹, John P Marino¹, Frank Delaglio¹, Luke W Arbogast¹, Ryan M Evans², Anthony Kearsley², Geneviève Gingras³, Houman Ghasriani³, Yves Aubin³, Gregory K Pierens⁴, Xinying Jia⁴, Mehdi Mobli⁴, Hamish G Grant⁵, David W Keizer⁵, Kristian Schweimer⁶, Jonas Ståhle⁷, Göran Widmalm⁷, Edward R Zartler⁸, Chad W Lawrence⁹, Patrick N Reardon⁹, John R Cort⁹, Ping Xu¹⁰, Feng Ni¹⁰, Saeko Yanaka¹¹, Koichi Kato¹¹, Stuart R Parnham¹², Desiree Tsao¹³, Andreas Blomgren¹⁴, Torgny Rundlöf¹⁴, Nils Trieloff¹⁵, Peter Schmieder¹⁵, Alfred Ross¹⁶, Ken Skidmore¹⁷, Kang Chen¹⁸, David Keire¹⁸, Darón I Freedberg¹⁹, Thea Suter-Stahel²⁰, Gerhard Wider²⁰, Gregor Ilc^{21

22}, Janez Plavec^{21

22}, Scott A Bradley²³, Donna M Baldisseri²⁴, Mauricio Luis Sforça²⁵, Ana Carolina de Mattos Zeri²⁶, Julie Yu Wei²⁷, Christina M Szabo²⁸, Carlos A Amezcua²⁸, John B Jordan²⁹, Mats Wikström³⁰

Affiliations

¹ a Institute of Bioscience and Biotechnology Research , National Institute of Standards and Technology and the University of Maryland , Rockville , MD , USA.
² b Applied and Computational Mathematics Division , National Institute of Standards and Technology , Gaithersburg , MD , USA.
³ c Centre for Biologics Evaluation, Biologics and Genetic Therapies Directorate , Health Canada , Ottawa , ON , Canada.
⁴ d The Centre for Advanced Imaging , The University of Queensland , St Lucia , QLD , Australia.
⁵ e Bio21 Molecular Science & Biotechnology Institute , The University of Melbourne , Victoria , Australia.
⁶ f Lehrstuhl Biopolymere , Universitaet Bayreuth , Bayreuth , Germany.
⁷ g Department of Organic Chemistry , Arrhenius Laboratory, Stockholm University , Stockholm , Sweden.
⁸ h Analytical R&D , Pfizer Essential Health , Lake Forest , IL , USA.
⁹ i Pacific Northwest National Laboratory , Earth and Biological Sciences Directorate , Richland , DC , USA.
¹⁰ j Department of Downstream Processing and Analytics , Human Health Therapeutics Research Centre, National Research Council of Canada , Montreal , Quebec , Canada.
¹¹ k Institute for Molecular Science and Exploratory Research Center on Life and Living Systems , National Institutes of Natural Sciences , Myodaiji, Okazaki , Japan.
¹² l Department of Biochemistry and Molecular Biology , Medical University of South Carolina , Charleston , SC , USA.
¹³ m Analytical Development , Momenta Pharmaceuticals , Cambridge , MA , USA.
¹⁴ n Laboratory Unit , Swedish Medical Products Agency, Laboratory , Uppsala , Sweden.
¹⁵ o NMR-supported Structural Biology , Leibniz-Forschungsinstitut für Molekulare Pharmakologie (FMP) , Berlin , Germany.
¹⁶ p Roche Pharmaceutical Research & Early Development , Pre-Clinical CMC, Roche Innovation Center Basel , Basel , Switzerland.
¹⁷ q Analytical Operations , Genentech , South San Francisco , CA , USA.
¹⁸ r Center for Drug Evaluation and Research , Food and Drug Administration , Maryland , USA.
¹⁹ s Center for Biologics Evaluation and Research , Food and Drug Administration , Maryland , USA.
²⁰ t Department of Biology , Institute of Molecular Biology and Biophysics , ETH Zurich, Zurich , Switzerland.
²¹ u NMR Centre , EN-FIST Centre of Excellence , Ljubljana , Slovenia.
²² v NMR Centre , National Institute of Chemistry , Ljubljana , Slovenia.
²³ w Eli Lilly and Company , Lilly Corporate Center , Indianapolis , IN , USA.
²⁴ x MRS - Application Science , Bruker BioSpin Corporation , Billerica , MA , USA.
²⁵ y Brazilian Biosciences National Laboratory (LNBio), Brazilian Center for Research in Energy and Materials (CNPEM) , Campinas, Brazil.
²⁶ z Brazilian Synchrotron Light Laboratory (LNLS), Brazilian Center for Research in Energy and Materials (CNPEM) , Campinas, Brazil , CEP.
²⁷ aa Protein Product Development , Biogen Inc ., Cambridge , MA , USA.
²⁸ ab Baxter Pharmaceuticals R&D , Baxter Healthcare , Round Lake, IL , USA.
²⁹ ac Global Regulatory and R&D Policy , Amgen Inc ., Thousand Oaks , CA , USA.
³⁰ ad Higher Order Structure, Attribute Sciences , Amgen Inc ., Thousand Oaks , CA , USA.

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 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.

Keywords: NISTmAb; chemometrics; comparability; higher order structure; monoclonal antibody (mAb) therapeutics; nuclear magnetic resonance spectroscopy (NMR).

Publication types

Research Support, Non-U.S. Gov't
Research Support, U.S. Gov't, Non-P.H.S.

MeSH terms

Antibodies, Monoclonal / chemistry*
Biopharmaceutics / standards*
Humans
Laboratories / standards*
Magnetic Resonance Spectroscopy / methods*
Reproducibility of Results

Substances

Antibodies, Monoclonal

Grants and funding

NIST acknowledges the support by NIST Biomanufacturing Program and NIST and W.M. Keck Foundation for support of biomolecular NMR instrumentation. The work at Stockholm University was supported by a grant from the Knut and Alice Wallenberg Foundation. The Brazilian Center for Research in Energy and Materials acknowledge the support by the Brazilian Ministry of Science, Technology, Innovation and Communication (MCTIC). A portion of the research was performed using NMR instrumentation at EMSL, a DOE Office of Science User Facility sponsored by the Office of Biological and Environmental Research and located at Pacific Northwest National Laboratory. This work at Health Canada was supported by Government of Canada. The work at NRC Canada was supported by the NRC Biologics and Biomanufacturing Program and by the Department of Downstream Processing and Analytics (for Protein NMR instrumentation), for which this paper is assigned the NRCC Publication No. NRC_HHT_53378. The work at the University of Queensland was supported by the Australian Research Council and The National Health and Medical Research Council as well as the University of Queensland. BNSP (Biomolecular NMR Spectroscopy Platform) ETH Zürich, is acknowledged for providing equipment and infrastructure. The NMR facility used for this work is supported by the Office of the Vice President for Research at the Medical University of South Carolina. The NMR maintenance was supported in part by the Nanotechnology Platform Program (Molecule and Material Synthesis) and by the grants (JP25102001, JP25102008, JP15K21708 and JP15H02491 to K.K.; JP17H05893 and 18K14892 to S.Y.) from the Ministry of Education, Culture, Sports, Science and Technology (MEXT) of Japan.