Woods Amina S, Jackson Shelley N
Structural Biology Unit, Cellular Neurobiology Branch, National Institute on Drug Abuse Intramural Research Program, National Institutes of Health, Baltimore, MD, USA.
Methods Mol Biol. 2010;656:99-111. doi: 10.1007/978-1-60761-746-4_5.
Tissue profiling and imaging by MALDI mass spectrometry has allowed the direct analysis and localization of biomolecules in tissue. However, due to the in situ nature of this technique, the complexity of tissue, and the need for a chemical matrix in MALDI, the signal recorded can be extremely complex and difficult to assign. Combining ion mobility with matrix-assisted laser desorption/ionization is a very powerful technique for fast separation and analysis of biomolecules in complex mixtures (such as tissue and cells), as isobaric lipid, peptide, and oligonucleotide molecular ions are pre-separated in the mobility cell before mass analysis. Differences in drift time of as much as 30% are obtained in a timescale of hundreds of microseconds. Molecular ions of the same biochemical family fall along trend lines when plotted in 2D plots of mobility drift time as a function of m/z. In this chapter ion mobility MALDI-MS ability to analyze various biomolecules in tissue, that is, lipids and proteins, as well as its ability to separate species from all of the major phospholipid classes from tissue and extracts, the effects that radyl chain length, degree of unsaturation, head group composition have upon their ion's cross section in the gas phase, and how it can be used not only to distinguish them from other biochemical groups in a mixture but also to differentiate them from other lipid species will be illustrated.
通过基质辅助激光解吸电离质谱(MALDI-MS)进行组织剖析和成像,能够直接分析组织中的生物分子并对其进行定位。然而,由于该技术的原位性质、组织的复杂性以及MALDI中对化学基质的需求,所记录的信号可能极其复杂且难以解析。将离子淌度与基质辅助激光解吸/电离相结合,是一种用于快速分离和分析复杂混合物(如组织和细胞)中生物分子的强大技术,因为等压脂质、肽和寡核苷酸分子离子在质量分析之前会在淌度池中预先分离。在数百微秒的时间尺度内,可获得高达30%的漂移时间差异。当以淌度漂移时间作为m/z的函数绘制二维图时,同一生化家族的分子离子会沿着趋势线分布。在本章中,将阐述离子淌度MALDI-MS分析组织中各种生物分子(即脂质和蛋白质)的能力,以及从组织和提取物中分离所有主要磷脂类物质的能力,脂链长度、不饱和度、头部基团组成对其离子在气相中的截面的影响,以及它如何不仅能够将它们与混合物中的其他生化基团区分开来,还能将它们与其他脂质种类区分开来。