Kantisto BV, Baarn, The Netherlands.
Department of Chemistry, KTH Royal Institute of Technology, Division of Applied Physical Chemistry, Stockholm, Sweden.
Electrophoresis. 2023 Jan;44(1-2):96-106. doi: 10.1002/elps.202200144. Epub 2022 Nov 14.
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.
单克隆抗体（mAbs）已成为用于治疗各种疾病的一类重要的生物制药。在制造过程中对其进行定量分析非常重要。在本工作中，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 mAb 的浓度。概述了 CZE 方法开发规则，特别是讨论了各种毛细管涂层，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 涂层导致最稳定的电动流和最有效的峰形。该方法在 0.1-10mg/ml 的范围内进行了验证，线性范围为 0.08-1.3mg/ml，通过将后一浓度范围的样品在水中稀释 10 倍，扩展范围为 1-10mg/ml。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（0.08mg/ml）的精密度和准确度稍差，但仍在监测目的的范围内。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。
