Product‐related fragments in monoclonal antibodies (mAbs) can have a significant impact on the efficacy and safety of the product. Capillary electrophoresis sodium dodecyl sulfate (CE‐SDS) is a commonly used method for fragment quantification, but it has challenges in peak identification due to the inability to enrich components and the incompatibility of SDS with mass spectrometry (MS). This article presents a workflow for identifying peaks in CE‐SDS analysis. The workflow involves comparing the migration time of peaks with that of standards and utilizing MS analysis to identify fragments. By employing this innovative systematic workflow, we successfully identified the CE‐SDS impurity peaks of seven antibody products. Among them, four products exhibited characteristic fragments associated with disulfide bonds (light chain [LC], heavy–light [HL] chain, heavy–heavy [HH] chain, and HH–LC) and a glycosylation‐related fragment non‐glycosylated heavy chain. Additionally, one product showed a fragment formed by the connection of HC_C130 and HC_C130, which is associated with a thioether bond. Furthermore, two other products displayed amino acid backbone breakage, with one product showing clipping at the HC region of A233–G285 and the other product showing clipping at the HC regions of A97–S158 and N342–T366. This workflow can be applied in early drug research, process development, or during the biologics license application stage to characterize fragments in therapeutic mAbs analyzed by CE‐SDS.

中文翻译：

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治疗性单克隆抗体的毛细管电泳十二烷基硫酸钠分析中检测到的片段的表征工作流程

单克隆抗体 (mAb) 中的产品相关片段会对产品的功效和安全性产生重大影响。毛细管电泳十二烷基硫酸钠（CE-SDS）是一种常用的片段定量方法，但由于无法富集组分且SDS与质谱（MS）不兼容，因此在峰识别方面面临挑战。本文介绍了在 CE-SDS 分析中识别峰的工作流程。该工作流程包括将峰的迁移时间与标准品的迁移时间进行比较，并利用 MS 分析来识别片段。通过采用这种创新的系统工作流程，我们成功鉴定了七种抗体产品的 CE-SDS 杂质峰。其中，四种产物表现出与二硫键相关的特征片段（轻链[LC]、重-轻[HL]链、重-重[HH]链和HH-LC）和糖基化相关片段非糖基化重链。链。此外，一种产品显示出由 HC_C 连接形成的片段130和HC_C130，它与硫醚键相连。此外，另外两种产品显示氨基酸主链断裂，其中一种产品显示 A 的 HC 区域被剪切233-G第285章另一个产品显示 A 的 HC 区域被剪裁97–S158和N第342章–T第366章。该工作流程可应用于早期药物研究、工艺开发或生物制剂许可申请阶段，以表征通过 CE-SDS 分析的治疗性单克隆抗体中的片段。