Department of Cell and Molecular Pharmacology and Experimental Therapeutics, Medical University of South Carolina, Charleston, SC, USA.
Adv Exp Med Biol. 2018;1104:59-76. doi: 10.1007/978-981-13-2158-0_4.
Matrix-assisted laser desorption ionization time-of-flight mass spectrometry (MALDI-TOF MS) has been used for two decades to profile the glycan constituents of biological samples. An adaptation of the method to tissues, MALDI mass spectrometry imaging (MALDI-MSI), allows high-throughput spatial profiling of hundreds to thousands of molecules within a single thin tissue section. The ability to profile N-glycans within tissues using MALDI-MSI is a recently developed method that allows identification and localization of 40 or more N-glycans. The key component is to apply a molecular coating of peptide-N-glycosidase to tissues, an enzyme that releases N-glycans from their protein carrier. In this chapter, the methods and approaches to robustly and reproducibly generate two-dimensional N-glycan tissue maps by MALDI-MSI workflows are summarized. Current strengths and limitations of the approach are discussed, as well as potential future applications of the method.
基质辅助激光解吸电离飞行时间质谱(MALDI-TOF MS)已经被用于分析生物样本中的聚糖成分二十年了。该方法的一种改进形式,即基质辅助激光解吸电离质谱成像(MALDI-MSI),可以在单个薄组织切片内高通量地对数百到数千种分子进行空间分析。利用 MALDI-MSI 对组织内的 N-聚糖进行分析是一种最近开发的方法,它可以识别和定位 40 种或更多的 N-聚糖。该方法的关键组成部分是在组织上应用肽-N-糖苷酶的分子涂层,这种酶可以将 N-聚糖从其蛋白质载体上释放出来。本章总结了通过 MALDI-MSI 工作流程稳健且可重复地生成二维 N-聚糖组织图谱的方法和方法。还讨论了该方法的当前优势和局限性,以及该方法的潜在未来应用。