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Online 2D-LC Analysis of Complex N-Glycans in Biopharmaceuticals Using the Agilent 1290 Infinity 2D-LC Solution
发布时间：
2019-05-15 15:13
来源：
安捷伦科技（中国）有限公司
| 样品： | 生物药物 | 项目： | 复杂 N-聚糖 |
| 方案文件名 | 下载 |
|---|---|
Online 2D-LC Analysis of Complex N-Glycans in Biopharmaceuticals Using the Agilent 1290 Infinity 2D-LC Solution |
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This Application Note presents different online 2D-LC concepts for the high-resolution analysis of complex N-linked glycans in erythropoietin (EPO) using the Agilent 1290 Infinity 2D-LC Solution with fluorescence detection. EPO has a complex glycosylation pattern with differently branched and charged glycans. A combination of hydrophilic interaction chromatography (HILIC) with weak anion exchange chromatography (WAX) enables highly orthogonal separation. Comprehensive 2D-LC analysis with HILIC in the first and WAX in the second dimension facilitates high-resolution 2D chromatography together with simultaneous charge profiling. In addition, multiple heart-cutting 2D-LC analysis, combining WAX and HILIC separation, provides a flexible alternative whereby the user can select multiple peaks to be analyzed in the second dimension and, moreover, run longer gradients.
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