尺寸排阻色谱法(SEC)是评估生物分子的产物纯度的基础分析方法。为了确保符合监管机构标准的准确和可重复的数据,用有效和连续的方法监测色谱柱是至关重要的。在这项研究中,使用在Regeneron制备的内部单克隆抗体评估19个SEC柱(WatersAcquityBEH200)。系统适用性参数(SSP)用于监测SEC测定的性能,包括USP决议,USP平板计数,USP拖尾因子,不对称因素,洗脱时间,峰宽,和峰值高度。建立了一般的线性模型,揭示了洗脱时间,峰宽,不对称因素,尾矿系数随注入次数的增加而增加,而峰高,决议,板数量减少。1000次注射后,拖尾系数和峰宽增加10%以上,而分辨率和板数从各自的起始值下降了10%以上。
Size exclusion chromatography (SEC) is a foundational analytical method to assess product purity of biological molecules. To ensure accurate and reproducible data that meet regulatory agency standards, it is critical to monitor the chromatographic column with efficient and continuous approaches. In this study, 19 SEC columns (Waters Acquity BEH200) were evaluated using an in-house monoclonal antibody made at Regeneron. System suitability parameters (SSPs) were used to monitor the performance of the SEC assay, including USP resolution, USP plate count, USP tailing factor, asymmetry factor, elution time, peak width, and peak height. A general linear model was built and revealed that elution time, peak width, asymmetry factor, and tailing factor increased with injection number, while peak height, resolution, and plate count decreased. After 1000 injections, tailing factor and peak width increased by more than 10%, while resolution and plate count decreased by more than 10% from their respective starting values.