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The Development of Novel Immunoassay System using fluorescent Rareearth Metal Complex Labels and Transparent Polymer Films.

使用荧光稀土金属络合物标签和透明聚合物薄膜开发新型免疫测定系统。

基本信息

批准号：
02555156
负责人：
YOTSUYANAGI Takao
金额：
$ 5.25万
依托单位：
Tohoku University
依托单位国家：
日本
项目类别：
Grant-in-Aid for Developmental Scientific Research (B)
财政年份：
1990
资助国家：
日本
起止时间：
1990 至 1991
项目状态：
已结题

项目摘要

This work has been conducted for the exploitation of time-resolved fluoimetric immunoassay systems by the combined use of long-lived rare-earth complex labels and transparent polymer films. The central aims of this work are ; (1) to establish the molecular designing criteria for the chelating ligands which give the long-lived 'energy transfer fluorescence' and (2) to examine plasticized polyvinyl chloride films as the media for the separation and the fluorescence enhancement. Main results are outlined as :(1) Time-resolved fluorescence measurement apparatus was accomplished wiht a pulsed nitrogen laser radiation source (337.1 nm). The accuracy and time-resolution capability of the builtup system was confirmed using the model luminescence system of benzoyltrifluoroacetonato Eu complex in ethanol.(2) The octadentate polyminocaboxylate ligands which have been Commercially available for calcium ion detection in biological systems have been proposed as promising candidates for the energy transfer fluorescence labels for the lanthanide ions. They have the 'antenna groups' of quinolyl or phenyl rings for absorption of near uv light and posses the N, O-donor set sa suffices for the coordination number of the lanthanide ions.(3) The strongly fluorescent chelate ion-pair Q[Eu(tta)_4] has been found to extracted in chloroform from alkaline solution at pH 10 (tta : thenoyltrifluoroacetonate, Q^+ : quaternary ammonium ion), whose fluorescence intensity was by a factor of 100 greater than that of the parent complex, [Eu(tta)_3].(4) A PVC film plasticized with dioctylphhalate was unfortunately not so beneficial to the fluorescence enhancement of Q[Eu(tta)_4], by contrast, it enhances the intensity of a triphenylmethane dye, ethyl violet. This observation indicates that the matching between fluorescent folutes and the film medium is a crucial factor to be conceived on the designing of such chemical systems for the luminescence in somewhat ambiguous nature.

本工作已进行开发的时间分辨荧光免疫分析系统的结合使用的长寿命的稀土络合物标记和透明的聚合物膜。本工作的主要目的是：（1）建立能产生长寿命“能量转移荧光”的螯合配体的分子设计标准;（2）研究增塑聚氯乙烯膜作为分离和荧光增强介质的作用。（1）采用脉冲氮激光（337.1nm）作为光源，研制了时间分辨荧光测量装置。用苯甲酰三氟丙酮铕配合物在乙醇中的模型发光体系验证了所建体系的准确性和时间分辨能力。(2)已商业化的用于生物体系中的钙离子检测的八齿聚氨基甲酸酯配体已被提出作为镧系离子的能量转移荧光标记物的有希望的候选者。它们具有喹啉基或苯环的“天线基团”，用于吸收近紫外光，并且N，O-供体组sa足以满足镧系离子的配位数。(3)在pH 10的碱性溶液中，用氯仿萃取得到了具有强荧光的螯合离子对Q[Eu（tta）_4]（tta：噻吩甲酰三氟丙酮，Q^+：季铵离子），其荧光强度是母体配合物[Eu（tta）_3]的100倍。(4)邻苯二甲酸二辛酯增塑的PVC膜对Q[Eu（tta）_4]的荧光增强作用不明显，相反，它增强了三苯甲烷染料乙基紫的荧光强度。这一观察结果表明，荧光液和薄膜介质之间的匹配是设计这种化学体系的一个关键因素，因为这种化学体系的发光性质有些模糊。