Motabar Dana, Kim Eunkyoung, Li Jinyang, Zhao Zhiling, Mouchahoir Trina, Gallagher D Travis, Schiel John E, Garige Mamatha, Sourbier Carole, Payne Gregory F, Bentley William E
Fischell Department of Bioengineering, University of Maryland, College Park, MD, USA.
Institute for Bioscience and Biotechnology Research, Rockville, MD, USA.
Nat Chem Biol. 2025 Feb;21(2):291-299. doi: 10.1038/s41589-024-01778-z. Epub 2024 Dec 2.
Protein function relies on sequence, folding and post-translational modification and molecular measurements are commonly used to reveal these structural features. Here, we report an alternative approach that represents these molecular features as readily measurable electronic patterns and validate this experimental approach by detecting structural perturbations commonly encountered during protein biomanufacturing. We studied a monoclonal antibody standard (from the National Institute of Standards and Technology) and focused on the electronic detection of variants that have undergone interchain disulfide bond reduction and methionine oxidation. Electronic detection of these structural perturbations is based on mediated electrochemical probing (MEP) that discerns patterns associated with the antibody's mediator-accessible redox activity. We demonstrate that MEP can rapidly (within minutes) and quantitatively detect alterations in the antibody's structural features and produce robust electronic signals that could enable monitoring of biomanufacturing processes. The ability to transduce information regarding a protein's structural perturbations into a more convenient electronic domain offers opportunities to apply the power of microelectronics and real-time data analytics to chemical and biological analysis.
蛋白质功能依赖于序列、折叠和翻译后修饰，分子测量通常用于揭示这些结构特征。在此，我们报告了一种替代方法，即将这些分子特征表示为易于测量的电子模式，并通过检测蛋白质生物制造过程中常见的结构扰动来验证这一实验方法。我们研究了一种单克隆抗体标准品（来自美国国家标准与技术研究院），并专注于对经历链间二硫键还原和甲硫氨酸氧化的变体进行电子检测。这些结构扰动的电子检测基于介导电化学探测（MEP），它能够识别与抗体可被介质接近的氧化还原活性相关的模式。我们证明，MEP能够快速（在数分钟内）并定量地检测抗体结构特征的变化，并产生强大的电子信号，从而实现对生物制造过程的监测。将有关蛋白质结构扰动的信息转换到更便捷的电子领域的能力，为将微电子技术和实时数据分析的力量应用于化学和生物分析提供了机会。
