腦脊髓液去除白蛋白與免疫球蛋白後於SELD/TOF質譜儀之蛋白質體學分析

CSF protein profiling by Surface-Enhanced Laser Desorption/Ionization Time-of Flight Mass Spectrometry after Depletion of Albumin and IgGs

腦脊髓液中的蛋白質可做為某些中樞神經系統疾病的診斷工具，但是，腦脊髓浪中有些量大的蛋白質卻常遮掩住某些量少的蛋白質的表現，而通常這些被遮推住的量少蛋白質就是重要的疾病生物標記。因此，發展一個成熟可信賴的技術來去除某些大的蛋白質、以保留量少蛋白質做為生物標記檢測，成為是否能應用蛋白質體學於這類研究的重要決定因素。我們應用抗人類自蛋白抗體，以及特別的交換樹脂來去除腦脊髓波中的白蛋白，再應用SELDI-TOF-MS，數小時內使得以成功分離出次蛋白質群。我們發展出的方法具高敏感度，而且相當快速省時，應可用為這類研究的有用常規工具。

Proteins in cerebrospinal fluid (CSF) could be used as indicators of some central nerve system diseases such as Alzheimer's disease. However, the high abundant proteins in CSF make the analysis very difficult because those proteins tend to shade those of lower abundance. Usually, some of the low abundant proteins are biomarkers. Therefore, it could be a better way to deprive high abundant albumin and lgGs from CSF first and analyze the remaining in order to find some imperceptible changes of low abundant proteins. We applied anti-human albumin antibody combine Protein A/G-Sepharose beads to specifically deprive albumin from the CSF sample. With high affinity to IgGs, Protein A/G could also deprive IgGs in the same approach. We perform single immunoprecipitation experiment and deprive albumin and lgGs from the CSF. SELDI-TOF-MS, combine high through-put separation and analysis of proteins, could profile the sub-proteome of a subject within hours. This system is an extremely sensitive and rapid method to analyze complex mixtures of proteins and peptides. The objective of this study was to utilize SELDI-TOF-MS to determine the specific protein markers of CSF. We setup the standard process procedure to reach the reliability and reproducibility of instrument output. Our results provide a rational basis for identifying biomarkers from CSF using SELDI- TOF-MS.