Antibodies 2017, 6(4), 25; https://doi.org/10.3390/antib6040025
Characterization of Monoclonal Antibody–Protein Antigen Complexes Using Small-Angle Scattering and Molecular Modeling
Maria Monica Castellanos 1,2, James A. Snyder 1, Melody Lee 1, Srinivas Chakravarthy 3, Nicholas J. Clark 4, Arnold McAuley 4 and Joseph E. Curtis 1,*
1
NIST Center for Neutron Research, National Institute of Standards and Technology, 100 Bureau Drive, Mail Stop 6102, Gaithersburg, MD 20899, USA
2
Institute for Bioscience and Biotechnology Research, 9600 Gudelsky Drive, Rockville, MD 20850, USA
3
Biophysics Collaborative Access Team-Sector 18ID, Illinois Institute of Technology, Advanced Photon Source, Argonne National Laboratory, Lemont, IL 60439, USA
4
Department of Drug Product Development, Amgen Incorporated, One Amgen Center Drive, Thousand Oaks, CA 91230, USA
*
Author to whom correspondence should be addressed.
Received: 16 November 2017 / Revised: 7 December 2017 / Accepted: 8 December 2017 / Published: 15 December 2017
(This article belongs to the Special Issue Monoclonal Antibodies)
Abstract
The determination of monoclonal antibody interactions with protein antigens in solution can lead to important insights guiding physical characterization and molecular engineering of therapeutic targets. We used small-angle scattering (SAS) combined with size-exclusion multi-angle light scattering high-performance liquid chromatography to obtain monodisperse samples with defined stoichiometry to study an anti-streptavidin monoclonal antibody interacting with tetrameric streptavidin. Ensembles of structures with both monodentate and bidentate antibody–antigen complexes were generated using molecular docking protocols and molecular simulations. By comparing theoretical SAS profiles to the experimental data it was determined that the primary component(s) were compact monodentate and/or bidentate complexes. SAS profiles of extended monodentate complexes were not consistent with the experimental data. These results highlight the capability for determining the shape of monoclonal antibody–antigen complexes in solution using SAS data and physics-based molecular modeling. View Full-TextKeywords:
antibody; antigen; small-angle scattering; docking; modeling; simulation; Monte Carlo
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Castellanos, M.M.; Snyder, J.A.; Lee, M.; Chakravarthy, S.; Clark, N.J.; McAuley, A.; Curtis, J.E. Characterization of Monoclonal Antibody–Protein Antigen Complexes Using Small-Angle Scattering and Molecular Modeling. Antibodies 2017, 6, 25.
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