高压、低温诱导的单克隆抗体展开和聚集：Fc 和 Fab 片段的作用,The Journal of Physical Chemistry B

当前位置： X-MOL 学术 › J. Phys. Chem. B › 论文详情

Our official English website, www.x-mol.net, welcomes your feedback! (Note: you will need to create a separate account there.)

高压、低温诱导的单克隆抗体展开和聚集：Fc 和 Fab 片段的作用
The Journal of Physical Chemistry B ( IF 2.8 ) Pub Date : 2022-06-08 , DOI: 10.1021/acs.jpcb.1c10528
Jordan E Berger ₁ , Susana C M Teixeira _{1,

2} , Kaelan Reed ₃ , Vladimir I Razinkov ₄ , Christopher J Sloey ₄ , Wei Qi ₄ , Christopher J Roberts ₁

Affiliation

在这项工作中检查了高压和低温对两种不同单克隆抗体 (MAb) 稳定性的影响。荧光和小角中子散射用于监测压力的原位效应以推断三级结构的变化并表征易于聚集的中间体。在一系列压力/温度条件下，两种 MAb 都观察到不同程度的部分去折叠。傅里叶变换红外光谱也用于异位监测二级结构的变化。在升高的压力和零摄氏度以下的温度下孵育后，天然二级结构的保存与三级展开的程度和可逆性无关。压力和温度的几种组合也用于辨别分离的 Ab 片段（Fab 和 Fc）对展开和聚集的各自贡献。每种抗体的片段显示出显着不同的部分展开特征和可逆性。在所有情况下，完整 MAb 的稳定性与 Fc 或 Fab 片段稳定性之间没有简单的相关性，这表明与完整 MAb 展开和聚集行为之间存在复杂的关系。尽管如此，

"点击查看英文标题和摘要"

High-Pressure, Low-Temperature Induced Unfolding and Aggregation of Monoclonal Antibodies: Role of the Fc and Fab Fragments

The effects of high pressure and low temperature on the stability of two different monoclonal antibodies (MAbs) were examined in this work. Fluorescence and small-angle neutron scattering were used to monitor the in situ effects of pressure to infer shifts in tertiary structure and characterize aggregation prone intermediates. Partial unfolding was observed for both MAbs, to different extents, under a range of pressure/temperature conditions. Fourier transform infrared spectroscopy was also used to monitor ex situ changes in secondary structure. Preservation of native secondary structure after incubation at elevated pressures and subzero ° C temperatures was independent of the extent of tertiary unfolding and reversibility. Several combinations of pressure and temperature were also used to discern the respective contributions of the isolated Ab fragments (Fab and Fc) to unfolding and aggregation. The fragments for each antibody showed significantly different partial unfolding profiles and reversibility. There was not a simple correlation between stability of the full MAb and either the Fc or Fab fragment stabilities across all cases, demonstrating a complex relationship to full MAb unfolding and aggregation behavior. That notwithstanding, the combined use of spectroscopic and scattering techniques provides insights into MAb conformational stability and hysteresis in high-pressure, low-temperature environments.

更新日期：2022-06-08

点击分享查看原文

点击收藏

阅读更多本刊新发论文