Klyushnichenko V, Kula M R
Institute of Enzymetechnology, Heinrich-Heine University Düsseldorf, Jülich, Germany.
Electrophoresis. 1997 Oct;18(11):2019-23. doi: 10.1002/elps.1150181123.
The application of sodium dodecyl sulfate (SDS)-polymer capillary electrophoresis (CE) to the analysis of proteins of hybridoma cell culture supernatant is demonstrated. All steps of the developed analysis are shown and discussed. Following optimization of sample preparation and concentration, as well as of the mode of injection all main protein components were separated with high resolution and selectivity. Proteolytic degradation of IgG, due to microbial contamination, was observed on SDS-gel CE and capillary zone electrophoresis (CZE). The technique was applied to monitor the cell cultivation process and concentrations of main components are demonstrated. IgG was found to increase from initially 5 mg/L to 20 mg/L after 4 days and to > 130 mg/L after 14 days of cultivation. The techniques may be applied to other biotechnological processes for the production or purification of proteins.
本文展示了十二烷基硫酸钠（SDS）-聚合物毛细管电泳（CE）在杂交瘤细胞培养上清液蛋白质分析中的应用。文中展示并讨论了所开发分析方法的所有步骤。在优化样品制备、浓缩以及进样模式后，所有主要蛋白质成分均以高分辨率和选择性得到分离。在SDS凝胶CE和毛细管区带电泳（CZE）上观察到由于微生物污染导致的IgG蛋白水解降解。该技术被应用于监测细胞培养过程，并展示了主要成分的浓度。发现IgG在培养4天后从最初的5 mg/L增加到20 mg/L，在培养14天后增加到>130 mg/L。该技术可应用于其他蛋白质生产或纯化的生物技术过程。
