Characterization of the NISTmAb Reference Material using Small-Angle Scattering and Molecular Simulation Part I: Dilute Protein Structures

Published

March 1, 2018

Author(s)

Maria Monica Castellanos Mantilla, Steven C Howell, David T. Gallagher, Joseph E. Curtis

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. In these series of papers on small-angle scattering, we characterize the IgG1 NISTmAb reference material in its histidine formulation buffer and study the higher order structure and interactions of proteins under a range of conditions. Part I presents the analysis of experimental scattering data with atomistic molecular simulations to characterize the monodisperse dilute solution of NISTmAb. 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 study shows that the scattering profile of the NISTmAb can be described using ensembles of flexible structures that explore a wide configuration space. The NISTmAb is flexible in solution with no preferred configuration or orientation of Fc and Fab domains with an average Rg of 49 angstrom} representing an ensemble of structures.

Citation

Analytical and Bioanalytical Chemistry

Volume

410

Issue

8

Pub Type

Journals

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Keywords

protein, small-angle scattering, SANS, molecular dynamics, Monte Carlo, antibody

Citation

Castellanos Mantilla, M. , Howell, S. , Gallagher, D. and Curtis, J. (2018), Characterization of the NISTmAb Reference Material using Small-Angle Scattering and Molecular Simulation Part I: Dilute Protein Structures, Analytical and Bioanalytical Chemistry, [online], https://tsapps.nist.gov/publication/get_pdf.cfm?pub_id=923805 (Accessed September 10, 2024)