Structure and dynamics of flexibly linked multi-domain proteins determined using spins, scattering, and simulations

Published

January 13, 2025

Author(s)

Veronika Szalai, Christina Bergonzo, Rachel Lyon, Zvi Kelman, Thomas Schmidt, Alexander Grishaev

Abstract

Antibody-based pharmaceuticals are the leading biologic drug platform ($50B/year). Despite a wealth of information collected on them, there is still a lack of knowledge on their inter-domain structural distributions, which impedes innovation and development. To address this measurement gap, we have designed a new methodology to derive biomolecular structure ensembles from distance distribution measurements via a library of tagged proteins bound to a non-isotopically labeled or otherwise modified target biologic. We have employed the NIST monoclonal antibody (NISTmAb) reference material as our development platform for use with spin-labeled affinity protein (SLAP) reagents. Using double electron-electron resonance (DEER) spectroscopy, we have determined point-to-point inter-spin distance distributions in spin-labeled protein complexes of both the isolated Fc domain and intact NISTmAb. Our SLAP reagents are a general and extendable technology, compatible with any non-isotopically labeled immunoglobulin G class mAb. Integrating molecular simulations with the DEER and solution X-ray scattering measurements, we enable simultaneous determination of structural distributions and dynamics of mAb-based biologics.

Citation

ChemMedChem

Pub Type

Journals

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https://doi.org/10.1002/cmdc.202400917

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Keywords

monoclonal antibody, double electron electron resonance spectroscopy, X-ray scattering, molecular simulation, molecular dynamics

Citation

Szalai, V. , Bergonzo, C. , Lyon, R. , Kelman, Z. , Schmidt, T. and Grishaev, A. (2025), Structure and dynamics of flexibly linked multi-domain proteins determined using spins, scattering, and simulations, ChemMedChem, [online], https://doi.org/10.1002/cmdc.202400917, https://tsapps.nist.gov/publication/get_pdf.cfm?pub_id=958073 (Accessed October 9, 2025)