介观流控-阀上实验室系统蛋白质定量分析的研究

Investigations on the Determination of Proteins in a Meso-fluidic Lab-on-valve System

在介观流控-阀上实验室系统中建立了蛋白质定量分析方法。在pH4．1的Britton-Robinson缓冲介质中，刚果红与蛋白质在室温下迅速结合，其生成产物在496nm处有最大吸收。用单因素法对实验参数进行了优化，确定最佳进样体积为20μL、试剂浓度为0．9g／L、用量4．0μL和检测流速为20μL／s。在最优条件下可对12．5～200mg／L之间的蛋白质进行准确定量分析，方法检出限为5．6mg／L，分析频率为60样／h。对人血清、尿样、乳饮料以及酸奶中总蛋白进行分析测定，测定结果与考马斯亮蓝方法一致。

The applications of a meso-fluidie lab-on-valve （LOV） system to the spectrophotometric determination of protein were investigated. Protein species in the sample solution reacts rapidly with Congo red at pH 4. 1 which forms a complex with a maximum absorption wavelength at 496 nm. A uni-variant approach was adopted for the optimization of the experimental parameters. A sample volume of 20μL was used along with 4.0μL of Congo red solution of 0.9 g/L, and a flow rate for the detection process of 20μL/s was employed. Under the optimal conditions, a linear calibration curve was obtained in the range of 12.5-200 mg/L BSA, along with a detection limit （3σ） of 5.6 mg/L and a sampling frequency of 60 h^-1. Protein concentrations in human serums, urine, milk and yoghourt were deterufined using this procedure and satisfactory agreements were obtained with the coomassie brilliant blue method.