在染色体时间尺度上探索电荷检测质谱
Lisa Strasser1, Florian Füssl1, Tomos E Morgan1,2
1Characterisation and Comparability Laboratory, NIBRT - the National Institute for Bioprocessing Research and Training, Foster Avenue, Mount Merrion, Blackrock Co, Dublin A94 X099, Ireland.
电荷检测质谱 (CDMS) 现在可以通过在线液态色谱进行. 这一进步允许快速准确地确定复杂蛋白质混合物的质量,即使获得时间短.
科学领域:
- 分析化学 分析化学
- 生物化学 生物化学
- 质谱测量质量谱测量
背景情况:
- 电荷检测质谱仪 (CDMS) 直接测量离子电荷和质量电荷比.
- 对于复杂的样品,如异质宏分子和混合物,在其他方法失败的情况下,CDMS是有价值的.
- 目前的CDMS方法通常需要很长的采集时间 (几十分钟到几个小时).
研究的目的:
- 调查CDMS分析在较短的时间尺度上的可行性,与液态染色学相兼容.
- 对复杂的生物样本进行在线液态染色学合CDMS的性能评估.
主要方法:
- 配合尺寸排除色谱 (SEC) 使用CDMS进行在线分离和质量分析.
- 分析转激素和β-银酸酶的混合物,然后进行复杂的葡萄糖蛋白和Fc融合蛋白的表征.
- 建立CDMS光谱数量与质量精度之间的相关性.
主要成果:
- 通过单次染色学运行的CDMS光谱 (每种蛋白质140-180光谱) 成功识别了转移素和β-银酸酶.
- 证明适用于复杂样品:α-1-酸甘氨蛋白和乙素被确定为染色体峰幅为~3分钟 (~200个光谱).
- 建立了CDMS光谱计数和可实现的质量精度之间的关系.
结论:
- CDMS与在线液态染色学兼容,使复杂混合物的快速分析成为可能.
- 这种带连线的技术显著减少了分析时间,同时保持了对蛋白质的确定性.
- 该方法适用于表征高度复杂的糖蛋白和融合蛋白,扩大CDMS应用.
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