CAJ | 학술논문

构建了一种在线阵列式二维常规柱液相色谱系统，并将其应用于分离血浆中的完整蛋白质。该系统以1根强阴离子交换柱作为第一维分离柱，8根阵列式反相色谱柱作为第二维分离柱。强阴离子交换柱分离的馏分通过十通阀被依次转移到第二维预柱上并得到保留富集，随后第二维流动相通过分流器同时将预柱上的蛋白质反冲至分析柱上进行分离。二维之间以及第二维阵列色谱柱之间均相互独立，从而可以提高系统分离的通量和总峰容量。采用该系统对血浆中的蛋白质进行了完整蛋白质水平上的分离。该系统具有高通量和高分辨率的特点，为血浆样品中高丰度蛋白质的去除以及血浆样品的深入研究提供了一种有效的手段。
구건료일충재선진렬식이유상규주액상색보계통，병장기응용우분리혈장중적완정단백질。해계통이1근강음리자교환주작위제일유분리주，8근진렬식반상색보주작위제이유분리주。강음리자교환주분리적류분통과십통벌피의차전이도제이유예주상병득도보류부집，수후제이유류동상통과분류기동시장예주상적단백질반충지분석주상진행분리。이유지간이급제이유진렬색보주지간균상호독립，종이가이제고계통분리적통량화총봉용량。채용해계통대혈장중적단백질진행료완정단백질수평상적분리。해계통구유고통량화고분변솔적특점，위혈장양품중고봉도단백질적거제이급혈장양품적심입연구제공료일충유효적수단。
Human plasma is one of the proteins-containing samples most difficult to character-ize on account of the wide dynamic concentration range of its intact proteins. Herein,we devel-oped a high-throughput conventional array-based two-dimensional liquid chromatographic sys-tem 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 trans-ferred into the corresponding reversed-phase liquid chromatographic precolumns for retention and enrichment using a 10-port electrically actuated multi-position valve. The second dimen-sional solvent flow was directly and identically split into 8 channels. The fractions were concur-rently back-flushed from the precolumns into the 8 conventional RP columns and were separa-ted 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 sep-aration. 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 pro-teins. The results indicated the established system is an effective method for removing high abundance proteins in plasma and in-depth research in plasma proteomics.