一种新的四溴荧光素-六氢吡啶磷光自组装环室温磷光标记试剂与人体疾病预报

A New Room Temperature Phosphorescence Labelling Reagent of Tetrabromofluorescein-piperidine Self-ordered Ring and Forecast of Diseases

发现四溴荧光素(HFinBr4)-六氢吡啶(Piperidine,P)在聚酰胺膜(PAM)的憎水性表面上能形成含多个HFinBr4发光分子的自组装环([HFinBr4)n-P-SOR,其中SOR是"self-ordered ring"的缩写)]与(HFinBr4)n-P-SOR的室温磷光(RTP)特性.以(HFinBr4)n-P-SOR标记麦胚凝集素(WGA)的产物(WGA-(HFinBr4)n-P-SOR)能与碱性磷酸酶(ALP)发生特异性的亲和吸附(AA)反应.生成的反应产物(ALP-WGA-(HFinBr4)n-P-SOR)能极大程度地提高生物靶上HFinBr4的分子数目,导致HFinBr4的RTP信号剧烈增强,比无形成(HFinBr4)n-P-SOR时的△I p增大7.8倍,据此建立了(HFinBr4)n-P-SOR亲和吸附固体基质室温磷光法(solid substrate-room temperature phosphorimetry,SS-RTP)测定痕量ALP的新方法(简称为(HFinBr4)n-P-SOR-AA-SS-RTP).该方法灵敏(LD为64.2 zg spot-1)、选择性好、简便、准确,用于人血清中痕量ALP含量的测定与人体疾病的预报,结果与酶联免疫法(ELISA)相吻合.同时,探讨了(HFinBr4)n-P-SOR-AA-SS-RTP测定ALP的反应机理.

It was discovered for the first time that tetrabromofluorescein（HFinBr4）-piperidine（P） could form a self-ordered ring（SOR） containing multiple luminescent molecules HFinBr4（which was called as（HFinBr4） n-P-SOR） on the hydrophobic surface of polyamide membrane（PAM）.At the same time,the room temperature phosphorescence（RTP） property of（HFinBr4） n-P-SOR was also firstly found.（WGA）-（HFinBr4） n-P-SOR,which was produced by（HFinBr4） n-P-SOR labeling triticum vulgare lectin（WGA）,could take specific affinity adsorption（AA） reaction with alkaline phosphatase（ALP）.And the product of AA reaction（ALP-WGA-（HFinBr4） n-P-SOR） could greatly increase the number of HFinBr4 molecules in the biology target leading to the RTP signal of HFinBr4 in（HFinBr4） n-P-SOR to enhance sharply.The ？Ip was 7.8 times higher than that of the system without（HFinBr4） n-P-SOR.According to the facts above,a new（HFinBr4） n-P-SOR affinity adsorption solid substrate-room temperature phosphorimetry（AA-SS-RTP） for determination of ALP was established（abbreviated to（HFinBr4） n-P-SOR-AA-SS-RTP）.The method was of high sensitivity（LD = 64.2 zg spot^-1）,good selectivity,high accuracy and convenience.It was used to determine the content of trace ALP in human serum and forecast human diseases,and the results coincided well with those obtained from enzyme-linked immunoassay（ELISA）.At the same time,the mechanism for determination of ALP by（HFinBr4） n-P-SOR-AA-SS-RTP was discussed.The academic thought that using（HFinBr4） n-P-SOR to label WGA to increase the number of HFinBr4 molecules in the biology target,could not only provide a new path to advance the sensitivity of SS-RTP,but also develop a new SS-RTP and technique of forecasting human diseases,which exploits the application field of SOR technology.