2′-脱氧胞苷探针测定血清样品中蛋白质含量

白蛋白在血浆总蛋白中占有很大的比例,高达40%~60%,它在人体内起着至关重要的作用,可以通过分析蛋白质的含量来反映人体的健康状况.人血清白蛋白表面具有多样化的结合位点,2′-脱氧胞苷能够与人血清白蛋白(HSA)相结合,但由于这种结合改变了HSA的结构,其荧光性质也就随之发生了改变.发明了一种新的测定蛋白质的方法,用2′-脱氧胞苷作荧光探针分子结合固定波长同步荧光光谱法对蛋白质的含量进行分析.在一定的条件下,当HSA的浓度满足在1.38~276μg·mL-1这一条件时,溶液的同步荧光强度(ISF)与HSA的浓度具有确定的线性关系.通过测定11份空白溶液发现检出限为0.024μg·mL-1(n=11).对实验条件进行了探究,发现溶液的pH值、扫描波长的变化量(Δλ)、试剂的加入顺序以及溶液的离子强度等因素都会影响体系荧光光谱特征及强度.还测量了人血清中的蛋白含量,回收率在98.4%~104.3%范围内.

Determination of Protein by Synchronous Fluorescence Technique with 2′-Deoxycytidine as Probe

The percentage of the albumin is large which reaches 40%-60%.It plays a vital role in the human body.The health of human body can be known by the content of protein.The binding sites on the surface of HSA are diversified,and2′-Deoxycytidine can react with human serum album.But the fluorescence have been altered because of the change on the structure of HSA.In this article,we reported a new method for the proteins determination by synchronous fluorescence spectrometry using 2′-Deoxycytidine as a fluorescence probe.The linear range of the synchronous fluorescence method was 1.38-262.2mg·L-1 with the detection limit 0.024μg·mL-1(n=11)for HSA.We found that the characterization and intensity of synchronous fluorescence spectral were related to the value ofΔλ,pH,ionic strength and so on by researching the experimental conditions.And the determination of the proteins in human serum samples was detected and the RSD was 98.4%-104.3%.