Westman A, Nilsson C L, Ekman R
Department of Psychiatry and Neurochemistry, Göteborg University, Sahlgrenska University Hospital/Mölndal, Sweden.
Rapid Commun Mass Spectrom. 1998;12(16):1092-8. doi: 10.1002/(SICI)1097-0231(19980831)12:16<1092::AID-RCM286>3.0.CO;2-N.
Matrix-assisted laser desorption/ionization time-of-flight mass spectra of proteins in cerebrospinal fluid analyzed without prior purification are presented. Less than 100 fmol amounts of proteins in the 10,000 to 20,000 u mass range and linked to human disease (multiple sclerosis, Alzheimer's disease, and stroke) were detected in a complex mixture of proteins and peptides, in the presence of high concentrations of salts, lipids and free amino acids. The mass resolution was sufficient to distinguish between the non-hydroxylated and hydroxylated forms of a 13,400 u protein. Simple fractionation of the cerebrospinal fluid using microbore-reversed phase high performance liquid chromatography improved signal-to-noise ratios in the mass spectra. High-accuracy peptide mass mapping and database searching were utilized to confirm the identity of several proteins. The presented results show that matrix-assisted laser desorption/ionization time-of-flight mass spectrometry could be used as a tool to perform rapid screening of chemically altered proteins in small volumes of biological fluids.
本文展示了未经预先纯化分析的脑脊液中蛋白质的基质辅助激光解吸/电离飞行时间质谱。在蛋白质和肽的复杂混合物中，在高浓度盐、脂质和游离氨基酸存在的情况下，检测到质量范围在10,000至20,000 u之间且与人类疾病（多发性硬化症、阿尔茨海默病和中风）相关的蛋白质含量低于100 fmol。质量分辨率足以区分13,400 u蛋白质的非羟基化和羟基化形式。使用微径反相高效液相色谱对脑脊液进行简单分级分离可提高质谱中的信噪比。利用高精度肽质量图谱和数据库搜索来确认几种蛋白质的身份。所呈现的结果表明，基质辅助激光解吸/电离飞行时间质谱可作为一种工具，用于对少量生物流体中化学改变的蛋白质进行快速筛选。
