Monoclonal antibodies (mAbs) have become an important class of biopharmaceuticals used for the treatment of various diseases. Their quantification during the manufacturing process is important. In this work, a capillary zone electrophoresis (CZE) method was developed for the monitoring of the mAb concentration during cell-culture processes. CZE method development rules are outlined, particularly discussing various capillary coatings, such as a neutral covalent polyvinyl alcohol coating, a dynamic successive multiple ionic-polymer coating, and dynamic coatings using background electrolyte additives such as triethanolamine (T-EthA) and triethylamine. The dynamic T-EthA coating resulted in most stable electro-osmotic flows and most efficient peak shapes. The method is validated over the range 0.1–10 mg/ml, with a linear range of 0.08–1.3 mg/ml and an extended range of 1–10 mg/ml by diluting samples in the latter concentration range 10-fold in water. The intraday precision and accuracy were 2%–12% and 88%–107%, respectively, and inter-day precision and accuracy were 4%–9% and 93%–104%, respectively. The precision and accuracy of the lowest concentration level (0.08 mg/ml) were slightly worse and still well in scope for monitoring purposes. The presented method proved applicable for analysing in-process cell-culture samples from different cell-culture processes and is possibly well suited as platform method.

中文翻译：

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有限样品制备的上游细胞培养过程样品中单克隆抗体定量监测的方法开发——各种毛细管涂层的评估

单克隆抗体 (mAb) 已成为一类重要的生物药物，用于治疗各种疾病。它们在制造过程中的量化很重要。在这项工作中，开发了一种毛细管区带电泳 (CZE) 方法，用于监测细胞培养过程中的 mAb 浓度。概述了 CZE 方法开发规则，特别讨论了各种毛细管涂层，例如中性共价聚乙烯醇涂层、动态连续多离子聚合物涂层，以及使用背景电解质添加剂（如三乙醇胺 (T-EthA) 和三乙胺）的动态涂层。动态 T-EthA 涂层产生最稳定的电渗流和最有效的峰形。该方法在 0.1–10 mg/ml 的范围内得到验证，线性范围为 0.08–1。3 mg/ml 和 1–10 mg/ml 的扩展范围，方法是将后者浓度范围内的样品在水中稀释 10 倍。日内精密度和准确度分别为 2%–12% 和 88%–107%，日间精密度和准确度分别为 4%–9% 和 93%–104%。最低浓度水平 (0.08 mg/ml) 的精密度和准确度稍差，但仍处于监测范围内。所提出的方法证明适用于分析来自不同细胞培养过程的过程中细胞培养样品，并且可能非常适合作为平台方法。最低浓度水平 (0.08 mg/ml) 的精密度和准确度稍差，但仍处于监测范围内。所提出的方法证明适用于分析来自不同细胞培养过程的过程中细胞培养样品，并且可能非常适合作为平台方法。最低浓度水平 (0.08 mg/ml) 的精密度和准确度稍差，但仍处于监测范围内。所提出的方法证明适用于分析来自不同细胞培养过程的过程中细胞培养样品，并且可能非常适合作为平台方法。