Citation: HUANG Zhi, HONG Guangfeng, GAO Mingxia, ZHANG Xiangmin. Development of online conventional array-based two-dimensional liquid chromatographic system for proteins separation in human plasma[J]. Chinese Journal of Chromatography, ;2014, 32(4): 343-348. doi: 10.3724/SP.J.1123.2013.10020 shu

Development of online conventional array-based two-dimensional liquid chromatographic system for proteins separation in human plasma

1.
1. Department of Chemistry, Fudan University, Shanghai 200433, China;

Corresponding author: ZHANG Xiangmin,
Received Date: 24 October 2013
Available Online: 18 December 2013
Fund Project: 国家高技术研究发展计划项目(2012AA020202)；科技部基础研究计划项目(2012CB910604)；国家自然科学基金项目(21027002，21175026，21275034). (2012AA020202)；科技部基础研究计划项目(2012CB910604)；国家自然科学基金项目(21027002，21175026，21275034)

Human plasma is one of the proteins-containing samples most difficult to characterize on account of the wide dynamic concentration range of its intact proteins. Herein, we developed a high-throughput conventional array-based two-dimensional liquid chromatographic system for proteins separation in human plasma in online mode. In the system, a conventional strong-anion exchange chromatographic column was used as the first separation dimension and eight parallel conventional reversed-phase liquid chromatographic columns were integrated as the second separation dimension. The fractions from the first dimension were sequentially transferred into the corresponding reversed-phase liquid chromatographic precolumns for retention and enrichment using a 10-port electrically actuated multi-position valve. The second dimensional solvent flow was directly and identically split into 8 channels. The fractions were concurrently back-flushed from the precolumns into the 8 conventional RP columns and were separated simultaneously. An 8-channel fraction collector was refitted to collect the reversed-phase liquid chromatographic fractions for further investigation. Bicinchoninic acid (BCA) dyeing solution was conveniently used for high-abundance protein location. Two separation dimensions were relatively independent parts, as well as each channel of the second dimensional array separation. Therefore, the new system could improve the separation throughput and total peak capacity. The system was successfully applied for the separation of human plasma intact proteins. The results indicated the established system is an effective method for removing high abundance proteins in plasma and in-depth research in plasma proteomics.
- array two dimensional chromatography,
- online,
- proteins,
- plasma,
- proteomics
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沈阳化工大学材料科学与工程学院沈阳 110142