微流控芯片电泳电导检测分离分析尿蛋白

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10 mmol/L硼砂溶液,电进样场强为300 V/cm,进样时间10 s,分离电压为600 V的实验条件下,10 min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0.8 g/L(S/N=3),在1-5 g/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 (HAS) and human transferrin (TRF) in urine samples from nephritic syndrome patients were separated on the designed microsystem, and HAS was quantitavely detected. Factors affecting on-chip electrophoresis processes, such as running buffer, separating voltage, injection modes and sampling time were inves-tigated and optimized. Under the optimized conditions of 10 mmol/L borax buffer (pH=10), 600 V separation voltage, and 300 V/cra electrical field strength and 10 s injection time, human serum albumin (HAS) and human transferring (TRF) were separated in 10 min with resolution of 2. 0. The detection limits of HAS and TRF were 0. 4 and 0. 8 g/L(S/N = 3), respectively. The correlation coef-ficients of the detection in the linear range of 1-5 g/L were at 0. 9478 and 0. 9491, respectively. The recoveries of HAS in the sample of patient urine were at 95. 4% -104. 1%.