Epitope screening using Hydrogen/Deuterium Exchange Mass Spectrometry (HDX-MS): An accelerated workflow for evaluation of lead monoclonal antibodies
Epitope screening using Hydrogen/Deuterium Exchange Mass Spectrometry (HDX-MS): An accelerated workflow for evaluation of lead monoclonal antibodies
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DOI：
10.1002/biot.202100358
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发表时间：
2021-11-21
影响因子：
4.7
通讯作者：
James, D. Andrew
中科院分区：
文献类型：
--
作者：
Zhu, Shaolong;Liuni, Peter;James, D. Andrew
背景表位作图是生物制品和疫苗开发的一个越来越重要的方面。治疗性抗体设计和生产的最新进展使得能够以快速增加的速率鉴定候选mAb，导致表征“结构”表位的显著瓶颈，这对于使用现有的高通量表位作图工具进行确定是具有挑战性的。在这里，引入了氢/氘交换质谱（HDX-MS）表位筛选工作流程，其非常适合于加速表征具有共同抗原的表位。主要方法和主要结果该方法在一组六种靶向Perglyn（PRN）的候选mAb上进行了演示。使用这种方法，在24小时总运行时间内使用两个HDX混合时间点明确确定了六个表位中的五个，这相当于使用常规工作流程映射单个表位所需的仪器时间。结论建立了一种快速的HDX-MS表位筛选流程。“筛选”工作流程成功地表征了PRN抗原上的五个（六个尝试中的）新表位;可用于支持疫苗抗原性测定的信息。
Background Epitope mapping is an increasingly important aspect of biotherapeutic and vaccine development. Recent advances in therapeutic antibody design and production have enabled candidate mAbs to be identified at a rapidly increasing rate, resulting in a significant bottleneck in the characterization of "structural" epitopes, that are challenging to determine using existing high throughput epitope mapping tools. Here, a Hydrogen/Deuterium Exchange Mass Spectrometry (HDX-MS) epitope screening workflow was introduced that is well suited for accelerated characterization of epitopes with a common antigen. Main methods and major results The method is demonstrated on set of six candidate mAbs targeting Pertactin (PRN). Using this approach, five of the six epitopes were unambiguously determined using two HDX mixing timepoints in 24 h total run time, which is equivalent to the instrument time required to map a single epitope using the conventional workflow. Conclusion An accelerated HDX-MS epitope screening workflow was developed. The "screening" workflow successfully characterized five (out of six attempted) novel epitopes on the PRN antigen; information that can be used to support vaccine antigenicity assays.