Molecular Insights into the Thermal Stability of mAbs with Variable-Temperature Ion-Mobility Mass Spectrometry
Molecular Insights into the Thermal Stability of mAbs with Variable-Temperature Ion-Mobility Mass Spectrometry
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DOI：
10.1002/cbic.201500574
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发表时间：
2016-01-01
期刊：
影响因子：
3.2
通讯作者：
Barran, Perdita E.
中科院分区：
文献类型：
--
作者：
Pacholarz, Kamila J.;Peters, Shirley J.;Barran, Perdita E.
以蛋白质为基础的治疗药物如单克隆抗体（mab）的聚集会影响治疗的效果，甚至会诱发对患者不利的效应。蛋白质工程是通过增加天然折叠的热力学稳定性来改变单抗容易聚集的状态，这可能反过来改变构象的灵活性。利用变温离子迁移率质谱技术（VT-IM-MS）对三种完整IgG分子和两种Fc-hinge片段的热稳定性进行了研究。我们观察到分离蛋白的构象随温度（300-550K）的变化。观察到的IgG亚类之间热稳定性的差异可以通过铰链区域减轻高阶结构组织的变化来合理化。VT-IM-MS提供了对单克隆抗体结构热力学的见解，并作为一种有前途的工具，用于治疗性蛋白质的热稳定性研究。
The aggregation of protein-based therapeutics such as monoclonal antibodies (mAbs) can affect the efficacy of the treatment and can even induce effects that are adverse to the patient. Protein engineering is used to shift the mAb away from an aggregation-prone state by increasing the thermodynamic stability of the native fold, which might in turn alter conformational flexibility. We have probed the thermal stability of three types of intact IgG molecules and two Fc-hinge fragments by using variable-temperature ion-mobility mass spectrometry (VT-IM-MS). We observed changes in the conformations of isolated proteins as a function of temperature (300-550K). The observed differences in thermal stability between IgG subclasses can be rationalized in terms of changes to higher-order structural organization mitigated by the hinge region. VT-IM-MS provides insights into mAbs structural thermodynamics and is presented as a promising tool for thermal-stability studies for proteins of therapeutic interest.