کد مقاله | کد نشریه | سال انتشار | مقاله انگلیسی | نسخه تمام متن |
---|---|---|---|---|
1216214 | 1494088 | 2014 | 5 صفحه PDF | دانلود رایگان |
• MSpak GF-4A precolumn is effective to selectively trap polysorbates on the precolumn in the on-line mode.
• HSA aggregates are in part trapped in the precolumn at lower temperature.
• r-HuEPO aggregates are in part trapped at higher temperatures.
• Setting the column temperature minimizes such inconvenient trapping of the aggregates.
Size-exclusion chromatography (SEC) is a useful method for quantification of protein aggregates because of its high throughput capacity and highly quantitative performance. One of the problems in this method concerns polysorbates, which are well-known additives for protein-containing products to prevent protein aggregation, but frequently interfere with the photometric detection of protein aggregates. We developed a new SEC method that can separate polysorbates from protein sample solutions in an on-line mode with a precolumn with size exclusion and reversed-phase mixed modes. The precolumn can effectively trap polysorbates in aqueous mobile phase, and the trapped polysorbates are easily eluted with acetonitrile-containing aqueous mobile phase to clean the precolumn. Small parts of protein aggregates may be also trapped on the precolumn depending on temperature and proteins. Setting appropriate column temperature can minimize such inconvenient trapping of aggregates.
Journal: Journal of Chromatography B - Volumes 953–954, 15 March 2014, Pages 68–72