分子探针N^6-羟基乙基腺苷同步荧光测定人血清中蛋白质

Determination of Proteins with Human Serum Albumin Using N^6-(2-hydroxyethyl)-adenosine as a Molecular Probe by Synchronous Fluorescence Technique

在模拟人体生理条件下,基于N6-羟基乙基腺苷(HEA)与人血清白蛋白(HSA)相互作用生成的复合物,导致血清白蛋白的内源荧光产生特异性变化,研究HEA-HSA体系的同步荧光光谱特征;同步荧光光谱特征及强度与Δλ值、反应温度等因素有关.在此基础上,建立了以HEA为探针测定蛋白质的新方法.在最佳实验条件下,HEA-HSA体系的同步荧光强度分别在0~331.2μg.mL-1和331.2~579.6μg.mL-1的浓度范围内与蛋白质浓度呈良好的线性关系,方法的检测限为4.43 ng.mL-1.实验结果表明:该法具有简单、快速、灵敏度高等优点,可直接用于人血清样品中蛋白质总量的测定,结果令人满意.

In simulative biological conditions, the interaction between N^6 -（2-hydroxyethyl）-adenosine（HEA） and human serum albumin （HSA） to form a complex（HEA-HSA）, resulting in a special change of the synchronous fluorescence of serum albumin. We have studied the characterization of synchronous fluorescence spectra of HEA-protein system with HEA as a probe, and the spectral characterization and intensity of synchronous fluorescence were related to the value of △λ and reaction temperature, and so on. On basis of these, the new method of determination of proteins was developed with HEA as a probe. Under the optimum experimental conditions, the synchronous fluorescence intensities of HEA-HSA was in good proportion to the concentration of HSA in the range of 0 - 331.2μg 2 mL^-1 and 331.2 - 579.6 μg · mL^-1 , respectively. The detection limit was 4.43 ng · mL^-1, Results suggested that the method possessed easy implementation, rapidity, high sensitivity, and so on, and the method was employed directly to determine the total proteins in serum albumin samples, and the results were satisfactory.