吖啶类荧光探针用于微量蛋白质测定的研究

Research into the application of acridine fluorescence probes to trace protein measurement

目的：利用合成的3种新型的蛋白质分子荧光探针N-（2-二甲氨基）乙基-9-氯吖啶-4-甲酰胺（NCAF）、9-[（N-2-二甲氨乙基）吖啶-4-甲酰胺]-α-丙氨酸（NAFA）和4,9-二[N-（2-二甲氨基）乙基]-9-吖啶胺-4-甲酰胺（DNAF）结合荧光发射光谱,建立了3种新的微量蛋白质测定方法。方法：通过建立适宜的NCAF-SDS（十二烷基硫酸钠）、NAFA-SDS和DNAF-SDS荧光猝灭体系,牛血清白蛋白（BSA）的加入对体系的荧光具有恢复作用,并随着BSA加入浓度的增大,荧光恢复程度逐渐增强,并在一定的浓度范围内呈线性关系,由此建立了用新型吖啶类荧光探针测定BSA的荧光分析新方法。结果：从NCAF、NAFA到DNAF测定线性范围分别为5.0×10-9~6.8×10^-7,9.0×10^-9~8.2×10^-8和5.0×10^-9~8.3×10^-7mol·L^-1;测定灵敏度（3σ/K）分别为1.1×10^-10,3.8×10^-10和1.0×10^-10mol·L^-1。结论：该测定方法具有良好的荧光响应和稳定性,为微量蛋白质定量分析提供了新的技术体系。

Objective： To establish three new ways to measure trace protein by utilizing 3 new synthetized types of protein molecule fluorescence probes, namely, NCAF, NAFA and DNAF and by combining with fluorescence emission spectrum. Method： First establish a proper fluorescence quenching system of NCAF-sodium dodecyl sulfate（SDS）, NAFA-SDS and DNAF-SDS, then add bovine serum albumin（BSA）to help restore the fluorescence of the system, and the higher the concentration of BSA added, the more the fluorescence is restored with linear relationship exhibited within certain concentration range, and thus we have established a new fluorescence analysis method of measuring BSA with new types of acridine fluorescence probes. Result： The effects of NCAF 、NAFA and DNAF are good with the measured linear range of 5.0 ×10^-9~6.8×10^-7,9.0×10^-9~8.2×10^-8 and 5.0×10^-9·8.3×10^-7mol·L^-1 respectively and the measurement sensitivity（3σ/K）of 1.1×10^-10,3.8×10^-10 and 1.0×10^-10 mol·L^-1 respectively. Conclusion： The said measuring method boasts good fluorescence response and stability, and thus offers a new technical system for the quantitative analysis of trace protein.