| 去除血浆中高丰度蛋白质的二维液相色谱体系的建立 |
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| 作者姓名： | Zhu S Zhang X Gao M Yan G Zhang X |
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| 作者单位： | 1. 复旦大学化学系, 上海 200433; 2. 复旦大学生物医学研究院, 上海 200032 |
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| 基金项目： | 国家重点基础研究发展规划(“973”)项目(No.2007CB914100); 国家自然科学基金项目(No.21027002); 上海市重点学科建设项目(B109) |
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| 摘 要： | 血浆中高丰度蛋白质的存在严重干扰低丰度蛋白质的检测,是困扰血浆蛋白质组学研究的技术瓶颈之一。针对这一热点问题,建立了一种二维液相色谱(强阴离子交换色谱-反相高效液相色谱)分离系统,对血浆中的高丰度蛋白质进行了色谱定位并进行去除。选择TSKgel SuperQ-5PW为第一维色谱分离柱,第二维色谱分离采用Jupiter C4柱,对第一维的馏分进行进一步的分离。通过梯度优化,血浆样品经过二维系统得到充分分离。第二维分离过程中从紫外信号强度高(215 nm,大于20 mAU)的峰中选择10个峰,利用液相色谱-串联质谱鉴定出32种高丰度蛋白质,包括人血清白蛋白、免疫球蛋白G等高丰度蛋白质。该体系为血浆中更多高丰度蛋白质的去除以及血浆蛋白质组学的更深入研究提供了重要思路。
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| 关 键 词： | 蛋白质组学 二维液相色谱 高丰度蛋白质 血浆 |
| 收稿时间： | 2011-07-15 |
Construction of a two-dimensional liquid chromatography separation system for high abundance proteins depletion in human plasma |
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| Zhu S,Zhang X,Gao M,Yan G,Zhang X.Construction of a two-dimensional liquid chromatography separation system for high abundance proteins depletion in human plasma[J].Chinese Journal of Chromatography,2011,29(9):837-842. |
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| Authors: | Zhu Shaochun Zhang Xueyang Gao Mingxia Yan Guoquan Zhang Xiangmin |
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| Institution: | 1. Department of Chemistry, Fudan University, Shanghai 200433, China; 2. Institute of Biomedical Science, Fudan University, Shanghai 200032, China |
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| Abstract: | High abundance proteins existing in human plasma severely impede the detection of low abundance proteins.This is one of the most difficult problems encountered in plasma proteomics research.We developed a two-dimensional liquid chromatography system with strong anion exchange chromatography-reversed-phase liquid chromatography(SAX-RPLC) for the extensive separation of plasma proteins and selective depletion of high abundance proteins.TSKgel SuperQ-5PW was selected as the first dimensional separation column ... |
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| Keywords: | proteomics two dimensional liquid chromatography high abundance protein plasma |
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