Bhagu Jamini, Anderson Lissa C, Grant Samuel C, Mohammadigoushki Hadi
Department of Chemical and Biomedical Engineering, FAMU-FSU College of Engineering, Tallahassee, FL 32310, USA; Center for Interdisciplinary Magnetic Resonance, National High Magnetic Field Laboratory, Florida State University, Tallahassee, FL 32310, USA.
Center for Interdisciplinary Magnetic Resonance, National High Magnetic Field Laboratory, Florida State University, Tallahassee, FL 32310, USA; Department of Chemistry and Biochemistry, Florida State University, Tallahassee FL 32306, USA.
J Pharm Sci. 2025 Apr;114(4):103685. doi: 10.1016/j.xphs.2025.01.020. Epub 2025 Jan 25.
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 partial unfolding, which can lead to the formation of proteinaceous aggregates. Using a combination of dynamic surface tensiometry and spatially resolved 1D H NMR spectroscopy, this study investigates if adsorption of a model IgG2a-κ mAb to the oil-water interface affects its structure. Localized NMR spectroscopy was performed using voxels of 375 µm, 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 (T) 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.
单克隆抗体（mAb）是一类重要的生物治疗药物，可用于治疗多种疾病，包括癌症、自身免疫性疾病或呼吸道疾病（如COVID-19）。然而，在其整个研发过程中，单克隆抗体都会接触到气-水或油-水界面，这可能会引发部分解折叠，进而导致蛋白质聚集体的形成。本研究结合动态表面张力测定法和空间分辨一维氢核磁共振光谱，探究一种模型IgG2a-κ单克隆抗体吸附至油-水界面是否会影响其结构。使用375µm的体素进行局部核磁共振光谱分析，体素逐渐靠近油-水界面。随着时间的推移，油-水界面处的动态界面张力逐渐降低，证实了单克隆抗体吸附至该界面。局部核磁共振光谱分析结果表明，虽然与单克隆抗体相关的化学共振峰数量和化学位移频率未受影响，但随着体素逐渐靠近油-水界面，会观察到谱线展宽。此外，对位于界面中心的一个体素测定了单克隆抗体分子的自旋-自旋（T）弛豫，结果表明在单克隆抗体的各个共振峰中弛豫受到不同程度的影响，这表明由于界面的存在，单克隆抗体分子的旋转受到限制。最后，位于界面中心的体素中，单克隆抗体的表观扩散系数低于本体单克隆抗体。这些结果表明，这种特定的单克隆抗体与界面附近的本体单克隆抗体相相互作用，并且可能会与之进行交换。因此，这些局部核磁共振技术有潜力探测和量化界面层附近单克隆抗体性质的变化。
