Use of a polybrene capillary coating in capillary electrophoresis for rapid analysis of hemoglobin variants with on-line detection via an ion trap storage reflectron time-of-flight mass spectrometer
Use of a polybrene capillary coating in capillary electrophoresis for rapid analysis of hemoglobin variants with on-line detection via an ion trap storage reflectron time-of-flight mass spectrometer
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DOI：
10.1021/ac970076m
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发表时间：
1997-07-01
影响因子：
7.4
通讯作者：
Lubman, DM
中科院分区：
文献类型：
--
作者：
Li, MX;Liu, L;Lubman, DM
在毛细管电泳（CE）中，聚凝胺毛细管涂层已用于血红蛋白变体的快速分析。聚凝胺毛细管涂层的使用允许充分的分离以解析在胰蛋白酶消化整个蛋白质时形成的大量消化产物，使得不需要血红蛋白α和β链的预先分离，通过CE获得的消化峰的分辨率足以使得即使单个氨基酸取代也可以使用UV吸收检测来容易地检测。通过毛细管电泳分离并使用连接到离子阱存储/反射器飞行时间装置（CE/ESI-IT/reTOF）的电喷雾电离进行在线检测，进一步分析消化物，其中突变体和正常血红蛋白的总离子电泳图和质谱的比较可以检测突变位点的存在。CE分离和质量分析通常可以在10-15分钟内完成。证明了CE/ESI-IT/reTOF系统检测快速分离的独特能力，这些分离具有可能低于1 s fwhm的窄峰。该系统的速度对于使用CE分离在短时间内分离的大量峰的分辨率至关重要。
A polybrene capillary coating in capillary electrophoresis (CE) has been used for rapid analysis of hemoglobin variant digests, The use of the polybrene capillary coating has allowed sufficient separation to resolve the large number of digest products formed upon tryptic digestion of the whole protein, so that prior separation of the hemoglobin alpha and beta chains is not required, The resolution of the digest peaks obtained by CE is sufficient so that even single amino acid substitutions can easily be detected using UV absorption detection. The digest is further analyzed by capillary electrophoresis separation with on-line detection using electrospray ionization interfaced to the ion trap storage/reflectron time of flight device (CE/ESI-IT/reTOF), where a comparison of the total ion electropherograms and mass spectra of the mutant and normal hemoglobins can detect the presence of a mutation site. The CE separation and mass analysis can be accomplished in typically 10-15 min. The unique capability of the CE/ESI-IT/reTOF system for detection of fast separations with narrow peaks that may be under 1 s fwhm is demonstrated. The speed of this system is essential for resolution of the large number of peaks that are separated in a short time duration using CE separations.