A CE-MALDI interface based on the use of prestructured sample supports
A CE-MALDI interface based on the use of prestructured sample supports
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DOI：
10.1021/ac0011888
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发表时间：
2001-04-15
影响因子：
7.4
通讯作者：
Gobom, J
中科院分区：
文献类型：
--
作者：
Johnson, T;Bergquist, J;Gobom, J
我们已经开发了一种离线耦合的毛细管电泳（CE）基质辅助激光解吸/电离飞行时间质谱（MALDI-TOF-MS）的基础上CE馏分收集到预结构的MALDI样品支持。使用标准肽混合物研究分析物残留和检测灵敏度。检测到低飞摩尔量，未发现明显的残留。用人胎脑cDNA表达文库中蛋白质的胰蛋白酶消化混合物评价了该方法的性能。与直接MALDI-MS分析相比，当使用CE-MALDI接口时，鉴定的肽的总数从47增加到211。对于不同蛋白质，CE-MALDI的序列覆盖率在25-60%范围内，对应于相对于粗混合物的MALDI-MS获得的序列覆盖率增加1.3-4.9倍。样品组分的分级也有助于通过MAG DI源后衰变分析进行蛋白质鉴定。我们的初步结果表明，这种CE-MALDI接口可用于分析复杂的肽混合物分离的生物组织。
We have developed an off-line coupling of capillary electrophoresis (CE) to matrix-assisted laser desorption/ ionization time-of-flight mass spectrometry (MALDI-TOF-MS) based on CE fraction collection onto prestructured MALDI sample supports. Analyte carryover and detection sensitivity were investigated using a standard peptide mixture. Low femtomole amounts were detected, and no noticeable carryover was discovered. The performance of the method was evaluated with a mixture of tryptic digests of proteins from a human fetal brain cDNA expression library. The total number of identified peptides was increased from 47 to 211 when the CE-MALDI interface was used compared to direct MALDI-MS analysis. Sequence coverage with CE-MALDI was in the 25-60% range for the different proteins, corresponding to an increase of 1.3-4.9 times relative to that obtained with MALDI-MS of the crude mixture. Fractionation of sample components also facilitated protein identification by MAG DI postsource decay analysis. Our initial results suggest this CE-MALDI interface can be used for the analysis of complex peptide mixtures isolated from biological tissues.