Mass Spectrometric-Based Stable Isotopic 2-Aminobenzoic Acid Glycan Mapping for Rapid Glycan Screening of Biotherapeutics
Mass Spectrometric-Based Stable Isotopic 2-Aminobenzoic Acid Glycan Mapping for Rapid Glycan Screening of Biotherapeutics
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DOI：
10.1021/ac902617t
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发表时间：
2010-02-15
影响因子：
7.4
通讯作者：
Cockrill, Steven L.
中科院分区：
文献类型：
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作者：
Prien, Justin M.;Prater, Bradley D.;Cockrill, Steven L.
在生物治疗产品生命周期的不同阶段，包括克隆选择、工艺改变和批次释放测试的质量控制，快速、灵敏、可靠的高水平葡聚糖筛选方法是必要的。传统的葡聚糖筛选涉及基于层析或电泳分离的方法，尽管这些方法可重复性，但这些方法可能很耗时。即使是工作在几十分钟时间尺度上的超高性能色谱和微流控集成LC/MS系统，当要分析数百个样品时，也会变得很长。相比之下，基于直接输注质谱仪(MS)的葡聚糖筛选方法在毫秒级的时间尺度上获取数据，表现出精致的灵敏度和重复性，并且易于自动峰注释。此外，还可以通过顺序质谱学同时进行多糖物种的表征。在这里，我们展示了一种使用稳定同位素2-氨基苯甲酸(2-AA)的定量高通量MS作图方法，用于快速筛选高水平的多糖。
Fast, sensitive, robust methods for "high-level" glycan screening are necessary during various stages of a biotherapeutic product's lifecycle, including clone selection, process changes, and quality control for lot release testing. Traditional glycan screening involves chromatographic or electrophoretic separation-based methods, and, although reproducible, these methods can be time-consuming. Even ultrahigh-performance chromatographic and microfluidic integrated LC/MS systems, which work on the tens of minute time scale, become lengthy when hundreds of samples are to be analyzed. Comparatively, a direct infusion mass spectrometry (MS)-based glycan screening method acquires data on a millisecond time scale, exhibits exquisite sensitivity and reproducibility, and is amenable to automated peak annotation. In addition, characterization of glycan species via sequential mass spectrometry can be performed simultaneously. Here, we demonstrate a quantitative high-throughput MS-based mapping approach using stable isotope 2-aminobenzoic acid (2-AA) for rapid "high-level" glycan screening.