Tiziani Stefano, Emwas Abdul-Hamid, Lodi Alessia, Ludwig Christian, Bunce Christopher M, Viant Mark R, Günther Ulrich L
Henry Wellcome Building for Biomolecular NMR Spectroscopy, CR UK Institute for Cancer Studies, University of Birmingham, Birmingham B15 2TT, UK.
Anal Biochem. 2008 Jun 1;377(1):16-23. doi: 10.1016/j.ab.2008.01.037. Epub 2008 Feb 5.
Blood serum is commonly used for clinical diagnostics because its protein composition bears a wealth of information about the health of an organism. More recently the analysis of the small molecule composition, the metabolome, has received increased attention because the metabolite composition is influenced by many diseases, by the administration of drugs and toxins, and by the diet and life style of an individual. When nuclear magnetic resonance spectroscopy is used as an analytical tool it is often preferable to remove catalytically active proteins, in particular for longer measurements, because metabolite concentrations are otherwise in constant flux. Here we have compared different protocols for the separation of proteins and metabolites, including precipitation methods and ultrafiltration. Whereas most extraction methods involving protein precipitation deplete some metabolites, ultrafiltration is superior in retaining metabolite concentrations and offers excellent reproducibility. We also describe a new method to recover the hydrophobic fraction for ultrafiltration with good reproducibility.
血清常用于临床诊断，因为其蛋白质组成承载着有关生物体健康状况的丰富信息。最近，小分子组成即代谢组的分析受到了更多关注，因为代谢物组成会受到许多疾病、药物和毒素的施用以及个体饮食和生活方式的影响。当使用核磁共振光谱作为分析工具时，通常最好去除具有催化活性的蛋白质，特别是对于较长时间的测量，因为否则代谢物浓度会不断变化。在这里，我们比较了分离蛋白质和代谢物的不同方案，包括沉淀方法和超滤。虽然大多数涉及蛋白质沉淀的提取方法会消耗一些代谢物，但超滤在保留代谢物浓度方面更具优势，并且具有出色的重现性。我们还描述了一种以良好重现性回收超滤疏水部分的新方法。
