离线2D-LC-MS系统的建立及其用于人肝蛋白质组学的研究

在蛋白质组学研究中，近年来提出的多维色谱-质谱联用技术有望成为继2D-PAGE-MS技术之后又一项重要的高通量技术平台，Yates，Davis，Wagner和Hancock等报道了二维强阳离子交换色谱-反相色谱-质谱(SCX-RPLC-MS)的在线联用分析系统，在该系统中，各维色谱的分离条件相互制约，只能从整体上加以平衡。

Establishment of Offline 2D-LC-MS System and Its Application to Human Liver Proteomics

A lab-made off line 2D-LC-MS system was developed for protein analysis. In case the off line 2D-LC was utilized for peptide separation, and the working conditions for strong cation exchange(SCX) chromatography and the succeeding reversed phase(RP) chromatography can be individually optimized, contrasted to the on-line system. The experimental results show that the mass fractions of acetonitrile for the SCX separation can be increased form 5% as references reported up to 20% so that the separation of peptides has been substantially improved. The off line system has been applied to the analysis of proteins extracted from the cancer tissue of nude mice inoculated by a human-hepatocellular-carcinoma cell-line D20. Some extreme basic proteins(pI>10) and larger proteins(M w>200 000), which are difficult to be isolated by the traditional 2D-PAGE method, have been identified in D20 sample.