Matrix-assisted ionization vacuum for protein detection, fragmentation and PTM analysis on a high resolution linear ion trap-orbitrap platform.
Matrix-assisted ionization vacuum for protein detection, fragmentation and PTM analysis on a high resolution linear ion trap-orbitrap platform.
复制标题
DOI：
10.1016/j.aca.2016.02.018
复制
发表时间：
2016-04-15
影响因子：
6.2
通讯作者：
Li L
中科院分区：
文献类型：
--
作者：
Chen B;Lietz CB;OuYang C;Zhong X;Xu M;Li L
基质辅助真空电离（MAIV）是一种新型的电离技术，它可以在真空中产生多电荷离子，而无需使用激光烧蚀或高压。MAIV可以在中真空和高真空基质辅助激光解吸/电离（MALDI）源和电喷雾电离（ESI）源中实现，而无需对仪器进行修改。在此，我们将MAIV适配到MALDI-LTQ-Orbitrap XL平台上用于生物分子分析。作为一种有吸引力的替代MALDI在溶液中和原位分析的生物分子，MAIV耦合到高分辨率和准确的质量（HRAM）MS仪器已成功地扩大了质量检测范围，并提高了由于产生多电荷离子的裂解效率。此外，MAIV的柔软性使得其在不稳定的翻译后修饰（PTM）分析中具有潜在的应用。在这项研究中，高达18.7 kDa的蛋白质被检测到多达18个电荷;完整的肽与不稳定的PTM在电离过程中得到很好的保存和表征MS/MS;复杂的组织样品中的肽和蛋白质进行检测和鉴定，无论是在液体提取物和原位。此外，我们证明，这种方法有助于MS/MS分析，与MALDI-MS/MS相比，具有更高的裂解效率。
Matrix-assisted ionization vacuum (MAIV) is a novel ionization technique that generates multiply charged ions in vacuum without the use of laser ablation or high voltage. MAIV can be achieved in intermediate-vacuum and high-vacuum matrix-assisted laser desorption/ionization (MALDI) sources and electrospray ionization (ESI) sources without instrument modification. Herein, we adapt MAIV onto the MALDI-LTQ-Orbitrap XL platform for biomolecule analysis. As an attractive alternative to MALDI for in solution and in situ analysis of biomolecules, MAIV coupling to high resolution and accurate mass (HRAM) MS instrument has successfully expanded the mass detection range and improved the fragmentation efficiency due to the generation of multiply charged ions. Additionally, the softness of MAIV enables potential application in labile post-translational modification (PTM) analysis. In this study, proteins as large as 18.7 kDa were detected with up to 18 charges; intact peptides with labile PTM were well preserved during the ionization process and characterized MS/MS; peptides and proteins in complex tissue samples were detected and identified both in liquid extracts and in situ. Moreover, we demonstrated that this method facilitates MS/MS analysis with improved fragmentation efficiency compared to MALDI-MS/MS.