微流控芯片电泳电导检测分离分析尿蛋白  

Separation and Detection of Urinary Proteins by Microfluidic Chip Integrated Contactless Conductivity Detector

作　　者：马亮波[1,2,3] 徐溢叫[1,2,3] 梁静[1] 刘海涛[2,3] 甘俊[1] 李栋顺[1,2] 彭金兰[1,2] 吴珊[1] 

机构地区：[1]重庆大学化学化工学院,重庆大学,重庆400030 [2]重庆大学新型微纳器件与系统技术国防重点学科实验室,重庆大学,重庆400030 [3]重庆大学微纳系统及新材料技术国际研发中心,重庆大学,重庆400030

出　　处：《分析化学》2011年第8期1123-1128,共6页Chinese Journal of Analytical Chemistry

基　　金：国家自然科学基金(No.20675089);全国博士学位论文作者专项资金(No.FSNEDD-200941);重庆市科技攻关项目(CSTC,No.2010AC5050);重庆大学研究生科技创新基金(No.CDJXS10-22-11-39);中央高校基本科研业务费(No.CDJXS1-22-00-05)资助项目

摘　　要：基于自行构建的微流控芯片电泳集成非接触式电导检测分析系统,建立了一种集进样、分离与检测为一体的微流控芯片电泳电导检测蛋白质的方法,并用于人白蛋白(HSA)和人转铁蛋白(TRF)两种尿蛋白的分离分析以及肾病综合症病人尿液中白蛋白的定量检测。考察并优化了缓冲液、分离电压、进样方式、进样时间等电泳分离的影响因素,在缓冲液为pH=10的10mmol/L硼砂溶液,电进样场强为300V/cm,进样时间10s,分离电压为600V的实验条件下,10min内完成了HSA和TRF的分离检测,分离度为2.0;检出限分别为0.4和0.8g/L(S/N=3),在1～5g/L线性范围内相关系数分别为0.9478和0.9491。实验中肾病综合症病人尿样中HSA的加标回收率为95.4%～104.1%。Based on the homemade analytical microsystem with the micro-fluidic chip integrated contactless conductivity detector,a new separating and detecting method of proteins on microchip with sample injection,separation,and detection was proposed.Human serum albumin（HSA） and human transferrin（TRF） in urine samples from nephritic syndrome patients were separated on the designed microsystem,and HSA was quantitavely detected.Factors affecting on-chip electrophoresis processes,such as running buffer,separating voltage,injection modes and sampling time were investigated and optimized.Under the optimized conditions of 10 mmol/L borax buffer（pH=10）,600 V separation voltage,and 300 V/cm electrical field strength and 10 s injection time,human serum albumin（HSA） and human transferring（TRF） were separated in 10 min with resolution of 2.0.The detection limits of HSA and TRF were 0.4 and 0.8 g/L（S/N=3）,respectively.The correlation coefficients of the detection in the linear range of 1-5 g/L were at 0.9478 and 0.9491,respectively.The recoveries of HSA in the sample of patient urine were at 95.4%-104.1%.

关 键 词：微流控芯片 集成电导检测 尿蛋白 电泳分离 

分 类 号：R446.1[医药卫生—诊断学]