Ar-gas cluster ion beam in ToF-SIMS for peptide and protein analysis
Ar-gas cluster ion beam in ToF-SIMS for peptide and protein analysis
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DOI：
10.1116/6.0000105
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发表时间：
2020-03-01
期刊：
影响因子：
2.1
通讯作者：
Lee, Tae Geol
中科院分区：
文献类型：
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作者：
Son, Jin Gyeong;Yoon, Sohee;Lee, Tae Geol
自从Ar气体团簇离子束（Ar-GCIBs）被引入到飞行时间二次离子质谱（ToF-SIMS）中以来，已经进行了各种尝试来分析需要低损伤分析和高灵敏度的有机材料和生物分子，因为Ar-GCIBs由于每个原子的低能量而允许大分子如肽和蛋白质的软电离。在这里，作者采用Ar-GCIB作为主要光束来检测蛋白质，包括人胰岛素，泛素和细胞色素C（分子量分别为5808，8564和12 327 Da）。他们已经证实，当Ar-GCIB用作初级离子束时，可以检测到完整的蛋白质。此外，他们通过分析肌红蛋白、细胞色素C和牛血清白蛋白中的胰蛋白酶消化肽，成功地鉴定了每种蛋白质。他们还尝试了表面酶消化来鉴定Si晶片表面上的蛋白质，并获得了与溶液中消化相同的结果。预计作者的表面消化方法可以使ToF-SIMS应用于分析生物组织中存在的蛋白质。
Since Ar-gas cluster ion beams (Ar-GCIBs) have been introduced into time-of-flight secondary ion mass spectrometry (ToF-SIMS), there have been various attempts to analyze organic materials and biomolecules that require low-damage analysis and high sensitivity, because Ar-GCIBs allow soft ionization of large molecules such as peptides and proteins due to the low energy per atom. Here, the authors adopted the Ar-GCIB as a primary beam to detect proteins including human insulin, ubiquitin, and cytochrome C (molecular weights are 5808, 8564, and 12 327 Da, respectively). They have confirmed that the detection of the intact proteins was possible when the Ar-GCIB was used as a primary ion beam. In addition, they successfully identified each protein by analyzing the trypsin-digested peptides in myoglobin, cytochrome C, and bovine serum albumin. They also attempted on-surface enzymatic digestion to identify proteins on the surface of the Si wafer and obtained results identical to those of in-solution digestion. It is expected that the authors' on-surface digestion method can enable the application of ToF-SIMS for the analysis of proteins present in biological tissues.