毛细管电泳-发光二极管诱导荧光检测法分析肝硬化及正常人血清蛋白质的差异

Analysis of difference of serum proteins between patients with cirrhosis and normal people by capillary electrophoresis with light-emitting diode induced fluorescence detection

应用毛细管电泳技术建立了人血清蛋白质毛细管电泳分析方法,对比了正常人与肝硬化患者血清蛋白质含量的差异。使用异硫氰酸荧光素(FITC)进行荧光标记,并考察光源与滤光片的波长、缓冲液的浓度、pH值、分离电压对分离效果的影响。建立了毛细管电泳-发光二极管诱导荧光法检测肝硬化患者血清及正常人血清蛋白质的方法;通过分离检测,获得了两者的蛋白质电泳谱图。经分析,在肝硬化患者的血清蛋白质中比在正常人的血清蛋白质中多检测到2个蛋白质峰,可能是和肝硬化发生相关的特异蛋白质;肝硬化患者与正常人的血清蛋白质电泳谱图的差异有统计学意义。该方法能实现人血清蛋白质的分离,可为临床诊断肝硬化做参考。

An experimental method of protein separation by capillary electrophoresis coupled with fight-emitting diode induced fluorescence detection （CE-LED-IF） was established. The flu- orescein isothiocyanate （FITC） was used as fluorescent dye in this method. An optical fiber was inserted into the separation capillary and was suited right at the detection window which was used to transmit the excitation light of LED. The efficiency of the light of LED and the detection limits of FITC was improved. The difference of serum proteins between the patients with cirrhosis and normal people was detected by this method. A series of effects on CE were studied, such as the wavelength of the light source and ~ter, the concentration and pH of the running buffer and the separation voltage. Then the optimized method was established for the determination of the serum proteins. The repeatability of CE-LED-IF was tested and the RSDs of migration time and peak area were 1.14% and 3.23%, respectively. The results showed that there were two specific peaks in cirrhosis serum. The differences between the patients with cir- rhosis and normal people having statistical significance indicated that these proteins might be the potential biomarkers for cirrhosis diagnosis. The developed CE-LED-IF method proved suit- able for the separation of serum proteins with good repeatability and supportable for the clinical diagnosis of cirrhosis.