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Energetic Dissection of Mab-Specific Reversible Self-Association Reveals Unique Thermodynamic Signatures

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Abstract

Purpose

Reversible self-association (RSA) remains a challenge in the development of therapeutic monoclonal antibodies (mAbs). We recently analyzed the energetics of RSA for five IgG mAbs (designated as A-E) under matched conditions and using orthogonal methods. Here we examine the thermodynamics of RSA for two of the mAbs that showed the strongest evidence of RSA (mAbs C and E) to identify underlying mechanisms.

Methods

Concentration-dependent dynamic light scattering and sedimentation velocity (SV) studies were carried out for each mAb over a range of temperatures. Because self-association was weak, the SV data were globally analyzed via direct boundary fitting to identify best-fit models, accurately determine interaction energetics, and account for the confounding effects of thermodynamic and hydrodynamic nonideality.

Results

mAb C undergoes isodesmic self-association at all temperatures examined, with the energetics indicative of an enthalpically-driven reaction offset by a significant entropic penalty. By contrast, mAb E undergoes monomer-dimer self-association, with the reaction being entropically-driven and comprised of only a small enthalpic contribution.

Conclusions

Classical interpretations implicate van der Waals interactions and H-bond formation for mAb C RSA, and electrostatic interactions for mAb E. However, noting that RSA is likely coupled to additional equilibria, we also discuss the limitations of such interpretations.

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Abbreviations

AUC:

analytical ultracentrifugation

DLS:

dynamic light scattering

mAb:

monoclonal antibody;

PBS:

phosphate buffered saline

RMSD:

root-mean-square deviation

RSA:

reversible self-association

SV:

sedimentation velocity

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Funding

This work was supported by MedImmune, LLC, now a member of AstraZeneca.

Author information

Author notes

  1. Mandi M. Hopkins

    Present address: Formulation Development, Regeneron Pharmaceuticals, Tarrytown, New York, 10591, USA

  2. Jared S. Bee

    Present address: Formulation and Drug Product Development, REGENXBIO Inc, Rockville, Maryland, 20850, USA

Authors and Affiliations

  1. Department of Pharmaceutical Sciences, University of Colorado Anschutz Medical Campus, Aurora, Colorado, 80045, USA

    Mandi M. Hopkins & David L. Bain

  2. Department of Dosage Form Design and Development, Biopharmaceuticals R&D, AstraZeneca, Gaithersburg, Maryland, 20878, USA

    Arun Parupudi & Jared S. Bee

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  1. Mandi M. Hopkins
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Correspondence to David L. Bain.

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Hopkins, M.M., Parupudi, A., Bee, J.S. et al. Energetic Dissection of Mab-Specific Reversible Self-Association Reveals Unique Thermodynamic Signatures. Pharm Res 38, 243–255 (2021). https://doi.org/10.1007/s11095-021-02987-0

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