等电聚焦
分辨率(逻辑)
色谱法
化学
分析化学(期刊)
毛细管作用
毛细管电泳
混合(物理)
生物系统
数学
计算机科学
材料科学
人工智能
物理
生物
复合材料
酶
量子力学
生物化学
作者
Julia Kahle,M. Stein,Hermann Wätzig
出处
期刊:Electrophoresis
[Wiley]
日期:2019-06-21
卷期号:40 (18-19): 2382-2389
被引量:14
标识
DOI:10.1002/elps.201900162
摘要
Abstract Capillary isoelectric focusing is indispensable for characterizing charge heterogeneity and isoelectric points of biopharmaceuticals. However, there are many influencing parameters and therefore method development is challenging. This study was performed to obtain an in‐depth understanding of the imaged CIEF methodology by applying a design of experiments approach. To describe the parameter's effects as objectively as possible, a polynomial regression model was derived for the most important responses. For this purpose, the reference monoclonal antibody suggested by the National Institute of Standards and Technology (NISTmAb) was used as test molecule. The total concentration and the mixing ratio of two types of carrier ampholytes and the added amounts of urea and l ‐arginine were selected as factors. The effects of these factors on 13 different responses such as resolution or p I values were investigated. In order to reduce the total number of experiments, a d ‐optimal design with 20 different parameter combinations and six replicates each was chosen. The most significant effects of the four factors were shown for the parameters related to separation efficiency and peak position. In addition, the extent of the factor's effect could be assessed. Depending on the selected factor combination, the p I value can differ up to approximately 0.15 p I units and the resolution value between main peak and adjacent basic peak can range from approximately 1.6 to 2.5, for example.
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