Characterization of the NISTmAb Reference Material using small-angle scattering and molecular simulation. Castellanos, M. M., Howell, S. C., Gallagher, D. T., & Curtis, J. E. Analytical and Bioanalytical Chemistry, 410(8):2141–2159, March, 2018. Place: Heidelberg Publisher: Springer Heidelberg WOS:000426466800009doi abstract bibtex Both conformational and colloidal stability of therapeutic proteins must be closely monitored and thoroughly characterized to assess the long-term viability of drug products. We characterized the IgG1 NISTmAb reference material in its histidine formulation buffer and report our findings on the higher order structure and interactions of NISTmAb under a range of conditions. In this paper we present the analysis of experimental small-angle scattering data with atomistic molecular simulations to characterize the monodisperse dilute solution of NISTmAb. In part II we describe the characterization of the NISTmAb at high protein concentration (Castellanos et al. 2018). The NISTmAb was found to be a flexible protein with a radius of gyration of 49.0 +/- 1.2 in histidine formulation buffer using a variety of neutron and X-ray scattering measurements. Scattering data were then modeled using molecular simulation. After building and validating a starting NISTmAb structure from the Fc and Fab crystallographic coordinates, molecular dynamics and torsion-angle Monte Carlo simulations were performed to explore the configuration space sampled in the NISTmAb and obtain ensembles of structures with atomistic detail that are consistent with the experimental data. Our results indicate that the small-angle scattering profiles of the NISTmAb can be modeled using ensembles of flexible structures that explore a wide configuration space. The NISTmAb is flexible in solution with no single preferred orientation of Fc and Fab domains, but with some regions of configuration space that are more consistent with measured scattering profiles. Analysis of inter-domain atomistic contacts indicated that all ensembles contained configurations where residues between domains are \textless= 4 , although few contacts were observed for variable and C (H) 3 regions.
@article{castellanos_characterization_2018,
title = {Characterization of the {NISTmAb} {Reference} {Material} using small-angle scattering and molecular simulation},
volume = {410},
issn = {1618-2642},
doi = {10.1007/s00216-018-0868-2},
abstract = {Both conformational and colloidal stability of therapeutic proteins must be closely monitored and thoroughly characterized to assess the long-term viability of drug products. We characterized the IgG1 NISTmAb reference material in its histidine formulation buffer and report our findings on the higher order structure and interactions of NISTmAb under a range of conditions. In this paper we present the analysis of experimental small-angle scattering data with atomistic molecular simulations to characterize the monodisperse dilute solution of NISTmAb. In part II we describe the characterization of the NISTmAb at high protein concentration (Castellanos et al. 2018). The NISTmAb was found to be a flexible protein with a radius of gyration of 49.0 +/- 1.2 in histidine formulation buffer using a variety of neutron and X-ray scattering measurements. Scattering data were then modeled using molecular simulation. After building and validating a starting NISTmAb structure from the Fc and Fab crystallographic coordinates, molecular dynamics and torsion-angle Monte Carlo simulations were performed to explore the configuration space sampled in the NISTmAb and obtain ensembles of structures with atomistic detail that are consistent with the experimental data. Our results indicate that the small-angle scattering profiles of the NISTmAb can be modeled using ensembles of flexible structures that explore a wide configuration space. The NISTmAb is flexible in solution with no single preferred orientation of Fc and Fab domains, but with some regions of configuration space that are more consistent with measured scattering profiles. Analysis of inter-domain atomistic contacts indicated that all ensembles contained configurations where residues between domains are {\textless}= 4 , although few contacts were observed for variable and C (H) 3 regions.},
language = {English},
number = {8},
journal = {Analytical and Bioanalytical Chemistry},
author = {Castellanos, Maria Monica and Howell, Steven C. and Gallagher, D. Travis and Curtis, Joseph E.},
month = mar,
year = {2018},
note = {Place: Heidelberg
Publisher: Springer Heidelberg
WOS:000426466800009},
keywords = {Antibody flexibility, Antibody structure, Higher order structure, NISTmAb reference material, Protein conformation, Small-angle scattering, aggregation mechanisms, antibody molecules, crystallography, dynamics, igg1, immunoglobulin, neutron-scattering, saxs, segmental flexibility, x-ray-scattering},
pages = {2141--2159}
}
Downloads: 0
{"_id":"xbpWYnwK6HBk6eEBm","bibbaseid":"castellanos-howell-gallagher-curtis-characterizationofthenistmabreferencematerialusingsmallanglescatteringandmolecularsimulation-2018","authorIDs":[],"author_short":["Castellanos, M. M.","Howell, S. C.","Gallagher, D. T.","Curtis, J. E."],"bibdata":{"bibtype":"article","type":"article","title":"Characterization of the NISTmAb Reference Material using small-angle scattering and molecular simulation","volume":"410","issn":"1618-2642","doi":"10.1007/s00216-018-0868-2","abstract":"Both conformational and colloidal stability of therapeutic proteins must be closely monitored and thoroughly characterized to assess the long-term viability of drug products. We characterized the IgG1 NISTmAb reference material in its histidine formulation buffer and report our findings on the higher order structure and interactions of NISTmAb under a range of conditions. In this paper we present the analysis of experimental small-angle scattering data with atomistic molecular simulations to characterize the monodisperse dilute solution of NISTmAb. In part II we describe the characterization of the NISTmAb at high protein concentration (Castellanos et al. 2018). The NISTmAb was found to be a flexible protein with a radius of gyration of 49.0 +/- 1.2 in histidine formulation buffer using a variety of neutron and X-ray scattering measurements. Scattering data were then modeled using molecular simulation. After building and validating a starting NISTmAb structure from the Fc and Fab crystallographic coordinates, molecular dynamics and torsion-angle Monte Carlo simulations were performed to explore the configuration space sampled in the NISTmAb and obtain ensembles of structures with atomistic detail that are consistent with the experimental data. Our results indicate that the small-angle scattering profiles of the NISTmAb can be modeled using ensembles of flexible structures that explore a wide configuration space. The NISTmAb is flexible in solution with no single preferred orientation of Fc and Fab domains, but with some regions of configuration space that are more consistent with measured scattering profiles. Analysis of inter-domain atomistic contacts indicated that all ensembles contained configurations where residues between domains are \\textless= 4 , although few contacts were observed for variable and C (H) 3 regions.","language":"English","number":"8","journal":"Analytical and Bioanalytical Chemistry","author":[{"propositions":[],"lastnames":["Castellanos"],"firstnames":["Maria","Monica"],"suffixes":[]},{"propositions":[],"lastnames":["Howell"],"firstnames":["Steven","C."],"suffixes":[]},{"propositions":[],"lastnames":["Gallagher"],"firstnames":["D.","Travis"],"suffixes":[]},{"propositions":[],"lastnames":["Curtis"],"firstnames":["Joseph","E."],"suffixes":[]}],"month":"March","year":"2018","note":"Place: Heidelberg Publisher: Springer Heidelberg WOS:000426466800009","keywords":"Antibody flexibility, Antibody structure, Higher order structure, NISTmAb reference material, Protein conformation, Small-angle scattering, aggregation mechanisms, antibody molecules, crystallography, dynamics, igg1, immunoglobulin, neutron-scattering, saxs, segmental flexibility, x-ray-scattering","pages":"2141–2159","bibtex":"@article{castellanos_characterization_2018,\n\ttitle = {Characterization of the {NISTmAb} {Reference} {Material} using small-angle scattering and molecular simulation},\n\tvolume = {410},\n\tissn = {1618-2642},\n\tdoi = {10.1007/s00216-018-0868-2},\n\tabstract = {Both conformational and colloidal stability of therapeutic proteins must be closely monitored and thoroughly characterized to assess the long-term viability of drug products. We characterized the IgG1 NISTmAb reference material in its histidine formulation buffer and report our findings on the higher order structure and interactions of NISTmAb under a range of conditions. In this paper we present the analysis of experimental small-angle scattering data with atomistic molecular simulations to characterize the monodisperse dilute solution of NISTmAb. In part II we describe the characterization of the NISTmAb at high protein concentration (Castellanos et al. 2018). The NISTmAb was found to be a flexible protein with a radius of gyration of 49.0 +/- 1.2 in histidine formulation buffer using a variety of neutron and X-ray scattering measurements. Scattering data were then modeled using molecular simulation. After building and validating a starting NISTmAb structure from the Fc and Fab crystallographic coordinates, molecular dynamics and torsion-angle Monte Carlo simulations were performed to explore the configuration space sampled in the NISTmAb and obtain ensembles of structures with atomistic detail that are consistent with the experimental data. Our results indicate that the small-angle scattering profiles of the NISTmAb can be modeled using ensembles of flexible structures that explore a wide configuration space. The NISTmAb is flexible in solution with no single preferred orientation of Fc and Fab domains, but with some regions of configuration space that are more consistent with measured scattering profiles. Analysis of inter-domain atomistic contacts indicated that all ensembles contained configurations where residues between domains are {\\textless}= 4 , although few contacts were observed for variable and C (H) 3 regions.},\n\tlanguage = {English},\n\tnumber = {8},\n\tjournal = {Analytical and Bioanalytical Chemistry},\n\tauthor = {Castellanos, Maria Monica and Howell, Steven C. and Gallagher, D. Travis and Curtis, Joseph E.},\n\tmonth = mar,\n\tyear = {2018},\n\tnote = {Place: Heidelberg\nPublisher: Springer Heidelberg\nWOS:000426466800009},\n\tkeywords = {Antibody flexibility, Antibody structure, Higher order structure, NISTmAb reference material, Protein conformation, Small-angle scattering, aggregation mechanisms, antibody molecules, crystallography, dynamics, igg1, immunoglobulin, neutron-scattering, saxs, segmental flexibility, x-ray-scattering},\n\tpages = {2141--2159}\n}\n\n","author_short":["Castellanos, M. M.","Howell, S. C.","Gallagher, D. T.","Curtis, J. E."],"key":"castellanos_characterization_2018","id":"castellanos_characterization_2018","bibbaseid":"castellanos-howell-gallagher-curtis-characterizationofthenistmabreferencematerialusingsmallanglescatteringandmolecularsimulation-2018","role":"author","urls":{},"keyword":["Antibody flexibility","Antibody structure","Higher order structure","NISTmAb reference material","Protein conformation","Small-angle scattering","aggregation mechanisms","antibody molecules","crystallography","dynamics","igg1","immunoglobulin","neutron-scattering","saxs","segmental flexibility","x-ray-scattering"],"downloads":0},"bibtype":"article","biburl":"https://api.zotero.org/groups/2482731/items?key=XwkQOKEdyfbIVwc7Z5lXI5wN&format=bibtex&limit=100","creationDate":"2020-05-02T23:23:31.672Z","downloads":0,"keywords":["antibody flexibility","antibody structure","higher order structure","nistmab reference material","protein conformation","small-angle scattering","aggregation mechanisms","antibody molecules","crystallography","dynamics","igg1","immunoglobulin","neutron-scattering","saxs","segmental flexibility","x-ray-scattering"],"search_terms":["characterization","nistmab","reference","material","using","small","angle","scattering","molecular","simulation","castellanos","howell","gallagher","curtis"],"title":"Characterization of the NISTmAb Reference Material using small-angle scattering and molecular simulation","year":2018,"dataSources":["TpEJtrMJLJpcLwjNA"]}