Analytical Sciences, Amgen, Inc., 4000 Nelson Rd., Longmont, Colorado 80503, USA.
Anal Chem. 2010 Feb 15;82(4):1498-508. doi: 10.1021/ac902617t.
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.
在生物治疗产品的生命周期的各个阶段，包括克隆选择、工艺变更和放行测试的质量控制，都需要快速、灵敏、稳健的“高级”聚糖筛选方法。传统的聚糖筛选涉及基于色谱或电泳分离的方法，虽然可重现，但这些方法可能很耗时。即使是基于数十分钟时间尺度的超高效色谱和微流控集成 LC/MS 系统，在需要分析数百个样本时也会变得冗长。相比之下，基于直接进样质谱 (MS) 的聚糖筛选方法可以在毫秒时间尺度上获取数据，具有出色的灵敏度和重现性，并且适合自动化峰注释。此外，通过顺序质谱对聚糖种类进行表征可以同时进行。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 映射方法，用于快速“高级”聚糖筛选。
