Sodium Dodecyl Sulfate Removal Interface to Enable Characterization of Fragment Impurities in Monoclonal Antibodies by Capillary Electrophoresis Sodium Dodecyl Sulfate Coupled to Mass Spectrometry

节目概要 单克隆抗体（mAb）已经成为免疫治疗中的主流治疗蛋白质。 生物制药行业（生物制药），以及对高效分析工具的需求， 特性和质量控制继续增加。在生产和储存过程中， mAb的一级结构可以由于不同的片段化诱导机制而发生，并且所产生的 片段种类可能对产品的安全性和有效性具有负面影响。因此，我们认为， 片段化是常规监测的关键质量属性，用于评估mAb的纯度和完整性 以及mAb衍生生物制品从生产到商业化。 十二烷基硫酸钠毛细管电泳（CE-SDS），这是毛细管 聚丙烯酰胺凝胶电泳（SDS-PAGE）的类似物，广泛用于监测mAb 在生产、配制、稳定性和商业放行过程中的片段。CE-SDS使用SDS来 使蛋白质变性并使非共价缔合的亚基可分离。分离是基于 电泳迁移由SDS结合诱导的表面电荷驱动，其与 蛋白质的分子量在筛分凝胶基质中，实现了基于尺寸的分离，因为所有的SDS- 蛋白质复合物具有相似的电荷尺寸比。CE-SDS提供了优异的片段分离度， 但是缺乏通过直接耦合到解析碎片的质谱的质量鉴定 目前在生物制药领域造成了很大的差距。 GMJ技术公司（GMJ）正在申请SBIR第一阶段资金，以开发微型SDS 在一些实施方案中，使用耗尽电泳（SDS）接口，以允许CE-SDS与质谱（MS）直接偶联。 所提出的接口将使用自由流动区带电泳机制，以去除SDS在线从 电喷雾电离质谱法（ESI-MS）前的目标CE-SDS峰。通过这项创新， GMJ旨在解决从研究到临床应用的mAb表征和质量控制的关键需求。 商业化通过消除几个费力的步骤，所提出的装置将显著改善 提高了mAb片段表征工作流程的效率，提高了分析精度， 灵敏度，并为生物制药研究人员提供节省成本的好处。

PROGRAM SUMMARY Monoclonal antibodies (mAbs) have become mainstream therapeutic proteins in the biopharmaceutical industry (biopharma), and the demand for efficient analytical tools for their characterization and quality control continues to increase. During manufacturing and storage, cleavage of mAb’s primary structure can occur due to different fragmentation induced mechanisms, and the resulting fragment species can have negative implication for the safety and efficacy of the product. Therefore, fragmentation is a critical quality attribute routinely monitored to assess the purity and integrity of mAbs and mAb-derived biologics from production to commercialization. Capillary electrophoresis with sodium dodecyl sulfate (CE-SDS), which is the capillary analogue of polyacrylamide gel electrophoresis (SDS-PAGE), is widely employed for monitoring mAb fragments during production, formulation, stability, and commercial release. CE-SDS employs SDS to denature proteins and render non-covalently associated subunits separable. The separation is based on electrophoretic migration driven by the surface charge induced by SDS binding, which is proportional to the protein’s molecular weight. In a sieving gel matrix, a size-based separation is achieved since all SDS- protein complexes have similar charge-to-size ratio. CE-SDS provides excellent resolution of fragments, but lack of mass identification through direct coupling to mass spectrometry of the resolved fragments currently creates a major gap in the biopharma. GMJ Technologies (GMJ) is requesting SBIR Phase I funding to develop a micro SDS depletion electrophoresis (SDE) interface to allow direct coupling of CE-SDS to mass spectrometry (MS). The proposed interface will use free-flow zone electrophoresis mechanism to remove SDS in-line from the CE-SDS peaks of interest prior to electrospray ionization mass spectrometry (ESI-MS). With this innovation, GMJ aims to address a critical need in the characterization and quality control of mAbs from research to commercialization. By eliminating several laborious steps, the proposed device will significantly improve the efficiency in the characterization workflow for mAbs’ fragments, improve the analysis precision and sensitivity, and provide cost-saving benefits for biopharma researchers.