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应用 iTRAQ 定量蛋白质组学技术分析系统性红斑狼疮患者外周血单个核细胞的比较蛋白质组学。

Comparative proteome analysis of peripheral blood mononuclear cells in systemic lupus erythematosus with iTRAQ quantitative proteomics.

机构信息

Key laboratory of Laboratory Medical Diagnostics, Ministry of Education, Chongqing Medical University, 1 Yi Xue Yuan Road, Chongqing 400016, People's Republic of China.

出版信息

Rheumatol Int. 2012 Mar;32(3):585-93. doi: 10.1007/s00296-010-1625-9. Epub 2010 Dec 1.

Abstract

To identify and quantify protein profiles from peripheral blood mononuclear cells (PBMC) of systemic lupus erythematosus (SLE) patients with isobaric Tagging for Relative and Absolute protein Quantification (iTRAQ)-based proteomic technology and to find differentially expressed proteins in SLE. PBMC were collected from patients of six stable SLE, six active SLE, six rheumatoid arthritis (RA), and six healthy donors. After protein extraction and concentration, the pooled protein content was labeled with iTRAQ reagents and then subjected to multiple chromatographic fractionation and tandem mass spectrometry. ProteinPilot™ 3.0 software and a database of IPI (International Protein Index) human 3.62 were used for database searching and statistical analysis. A total of 452 proteins were identified. Of these, 67 unique proteins were observed twofold or more alteration in levels across groups. The proteins determined support existing knowledge and uncover novel biomarker candidates. These results indicate that iTRAQ-based technology can serve as a useful aid for identification and quantification proteins from PBMC.

摘要

为了鉴定和量化系统性红斑狼疮(SLE)患者外周血单个核细胞(PBMC)中的蛋白质谱,我们采用基于同位素标记相对和绝对定量(iTRAQ)的蛋白质组学技术,并寻找 SLE 中的差异表达蛋白。我们从六名稳定的 SLE 患者、六名活动期 SLE 患者、六名类风湿关节炎(RA)患者和六名健康供体中收集 PBMC。在进行蛋白质提取和浓缩后,将混合的蛋白质含量用 iTRAQ 试剂标记,然后进行多次色谱分离和串联质谱分析。我们使用 ProteinPilot™ 3.0 软件和 IPI(国际蛋白质索引)人类 3.62 数据库进行数据库搜索和统计分析。总共鉴定到 452 种蛋白质。其中,67 种独特蛋白质在各实验组中发生了两倍或更多的水平改变。这些确定的蛋白质支持现有的知识,并揭示了新的生物标志物候选物。这些结果表明,基于 iTRAQ 的技术可以作为鉴定和量化 PBMC 中蛋白质的有用辅助手段。

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