Nuclear Magnetic Resonance Study of Monoclonal Antibodies Near an Oil-Water Interface

Fuente: arXiv
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Main Authors: Bhagu, Jamini, Anderson, Lissa C., Grant, Samuel C., Mohammadigoushki, Hadi
Format: Preprint
Published: 2025
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author Bhagu, Jamini
Anderson, Lissa C.
Grant, Samuel C.
Mohammadigoushki, Hadi
author_facet Bhagu, Jamini
Anderson, Lissa C.
Grant, Samuel C.
Mohammadigoushki, Hadi
contents Monoclonal antibodies (mAb) represent an important class of biologic therapeutics that can treat a variety of diseases including cancer, autoimmune disorders or respiratory conditions (e.g. COVID-19). However, throughout their development, mAb are exposed to air-water or oil-water interfaces that may trigger mAb partial unfolding that can lead to the formation of proteinaceous aggregates. Using a combination of dynamic surface tensiometry and spatially resolved 1D 1H NMR spectroscopy, this study investigates if adsorption of a model IgG2a-\k{appa} mAb to the oil-water interface affects its structure. Localized NMR spectroscopy was performed using voxels of 375 um, incrementally approaching the oil-water interface. Dynamic interfacial tension progressively decreases at the oil-water interface over time, confirming mAb adsorption to the interface. Localized NMR spectroscopy results indicate that, while the number of mAb-related chemical resonances and chemical shift frequencies remain unaffected, spectral line broadening is observed as voxels incrementally approach the oil-water interface. Moreover, the spin-spin (T2) relaxation of the mAb molecule was measured for a voxel centered at the interface and shown to be affected differentially across the mAb resonances, indicating a rotational restriction for mAb molecules due to presence of the interface. Finally, the apparent diffusion coefficient of the mAb for the voxel centered at the interface is lower than the bulk mAb. These results suggest that this specific mAb interacts with and may be in exchange with bulk mAb phase in the vicinity of the interface. As such, these localized NMR techniques offer the potential to probe and quantify alterations of mAb properties near interfacial layers.
format Preprint
id arxiv_https___arxiv_org_abs_2502_05563
institution arXiv
publishDate 2025
record_format arxiv
spellingShingle Nuclear Magnetic Resonance Study of Monoclonal Antibodies Near an Oil-Water Interface
Bhagu, Jamini
Anderson, Lissa C.
Grant, Samuel C.
Mohammadigoushki, Hadi
Biological Physics
Monoclonal antibodies (mAb) represent an important class of biologic therapeutics that can treat a variety of diseases including cancer, autoimmune disorders or respiratory conditions (e.g. COVID-19). However, throughout their development, mAb are exposed to air-water or oil-water interfaces that may trigger mAb partial unfolding that can lead to the formation of proteinaceous aggregates. Using a combination of dynamic surface tensiometry and spatially resolved 1D 1H NMR spectroscopy, this study investigates if adsorption of a model IgG2a-\k{appa} mAb to the oil-water interface affects its structure. Localized NMR spectroscopy was performed using voxels of 375 um, incrementally approaching the oil-water interface. Dynamic interfacial tension progressively decreases at the oil-water interface over time, confirming mAb adsorption to the interface. Localized NMR spectroscopy results indicate that, while the number of mAb-related chemical resonances and chemical shift frequencies remain unaffected, spectral line broadening is observed as voxels incrementally approach the oil-water interface. Moreover, the spin-spin (T2) relaxation of the mAb molecule was measured for a voxel centered at the interface and shown to be affected differentially across the mAb resonances, indicating a rotational restriction for mAb molecules due to presence of the interface. Finally, the apparent diffusion coefficient of the mAb for the voxel centered at the interface is lower than the bulk mAb. These results suggest that this specific mAb interacts with and may be in exchange with bulk mAb phase in the vicinity of the interface. As such, these localized NMR techniques offer the potential to probe and quantify alterations of mAb properties near interfacial layers.
title Nuclear Magnetic Resonance Study of Monoclonal Antibodies Near an Oil-Water Interface
topic Biological Physics
url https://arxiv.org/abs/2502.05563