Sundberg Mårten, Bergquist Jonas, Ramström Margareta
Department of Chemistry - BMC, Analytical Chemistry and Science for Life Laboratory, Uppsala University, Box 599, 751 24 Uppsala, Sweden.
Biochem Biophys Rep. 2015 Jul 23;3:68-75. doi: 10.1016/j.bbrep.2015.07.013. eCollection 2015 Sep.
As the number of fully sequenced animal genomes and the performance of advanced mass spectrometry-based proteomics techniques are continuously improving, there is now a great opportunity to increase the knowledge of various animal proteomes. This research area is further stimulated by a growing interest from veterinary medicine and the pharmaceutical industry. Cerebrospinal fluid (CSF) is a good source for better understanding of diseases related to the central nervous system, both in humans and other animals. In this study, four high-abundant protein depletion columns, developed for human or rat serum, were evaluated for dog CSF. For the analysis, a shotgun proteomics approach, based on nanoLC-LTQ Orbitrap MS/MS, was applied. All the selected approaches were shown to deplete dog CSF with different success. It was demonstrated that the columns significantly improved the coverage of the detected dog CSF proteome. An antibody-based column showed the best performance, in terms of efficiency, repeatability and the number of proteins detected in the sample. In total 983 proteins were detected. Of those, 801 proteins were stated as uncharacterized in the UniProt database. To the best of our knowledge, this is the so far largest number of proteins reported for dog CSF in one single study.
随着全序列动物基因组数量的不断增加以及基于先进质谱技术的蛋白质组学技术性能的持续提升，如今有一个绝佳的机会来增进对各种动物蛋白质组的了解。兽医学和制药行业日益浓厚的兴趣进一步推动了这一研究领域的发展。脑脊液（CSF）是深入了解人类和其他动物中枢神经系统相关疾病的良好来源。在本研究中，对四种为人类或大鼠血清开发的高丰度蛋白质去除柱进行了犬脑脊液评估。分析采用基于纳升液相色谱-线性离子阱-轨道阱串联质谱（nanoLC-LTQ Orbitrap MS/MS）的鸟枪法蛋白质组学方法。结果表明，所有选定的方法在去除犬脑脊液方面取得了不同程度的成功。结果证明，这些柱子显著提高了检测到的犬脑脊液蛋白质组的覆盖范围。就效率、重复性和样品中检测到的蛋白质数量而言，一种基于抗体的柱子表现最佳。总共检测到983种蛋白质。其中，801种蛋白质在UniProt数据库中被列为未表征蛋白质。据我们所知，这是迄今为止在一项单一研究中报道的犬脑脊液蛋白质数量最多的数据。
