摘要
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.
出处
《漳州师范学院学报(自然科学版)》
2010年第2期84-91,共8页
Journal of ZhangZhou Teachers College(Natural Science)
基金
福建省自然科学基金计划资助项目(2008J0313)
关键词
碱性磷酸酶
四溴荧光素-六氢吡啶-自组装环
亲和吸附固体基质室温磷光法
alkaline phosphatase
tetrabromofluorescein-piperidine self-ordered ring
affinity adsorption solid substrate-room temperature phosphorimetry