蛋白质荧光标记中协同效应的高效液相色谱  

Cooperative Effects in the Fluorescent Labeling of Proteins Studied by High-performance Liquid Chromatography

作　　者：乔晓强[1] 张琳[1] 梁振[1] 张维冰[1] 张丽华[1] 张玉奎[1] 

机构地区：[1]中国科学院大连化学物理研究所国家色谱研究分析中心,大连116023

出　　处：《高等学校化学学报》2007年第9期1657-1659,共3页Chemical Journal of Chinese Universities

基　　金：国家自然科学基金(批准号:20435020);中国科学院知识创新工程重要方向项目(批准号:KJCX2.YW.H09)资助.

摘　　要：To improve the detection sensitivity of low abundance proteins in samples,fluorescence labeling might be a good solution method.In this manuscript,with 5-({2-[(iodoacetyl)amino]ethyl} amino)naphthalene-1-sulfonic acid(1,5-I-AEDANS)as the derivatizing reagent,the cooperative effects of insulin and BSA during the derivatization were studied.Through HPLC analysis,it was found that with BSA added,the detection sensitivity of the derivatized insulin could be improved.When the concentration ratio of BSA to insulin reached 200∶1,the peak area of insulin was increased by over 4-fold,which demonstrates that the existence of BSA had a positive effect on the derivatization of insulin.Furthermore,even when the ratio reached 1000∶1,the S/N of insulin was 8,which could not be detected without BSA being added.However,on the contrary,with insulin being added,the peak area of the derivatized BSA was decreased,demonstrating that insulin had a negative effect on that of BSA.All these results might be of great significance to the detection of low abundance proteins in proteome study.To improve the detection sensitivity of low abundance proteins in samples, fluorescence labeling might be a good solution method. In this manuscript, with 5-（ {2-[ （iodoacetyl）amino] ethyl/ amino） naphthalene-l-sulfonic acid （1,5-I-AEDANS） as the derivatizing reagent, the cooperative effects of insulin and BSA during the derivatization were studied. Through HPLC analysis, it was found that with BSA added, the detection sensitivity of the derivatized insulin could be improved. When the concentration ratio of BSA to insu- lin reached 200： 1, the peak area of insulin was increased by over 4-fold, which demonstrates that the existence of BSA had a positive effect on the derivatization of insulin. Furthermore, even when the ratio reached 1000： 1, the S/N of insulin was 8, which could not be detected without BSA being added. However, on the contrary, with insulin being added, the peak area of the derivatized BSA was decreased, demonstrating that insulin had a negative effect on that of BSA. All these results might be of great significance to the detection of low abundance proteins in proteome study.

关 键 词：荧光标记 蛋白质 协同效应 高效液相色谱 

分 类 号：O657[理学—分析化学]