Development of Sensitive "Micro-immunoassay System" and Application to Determination of Cancer Marker

灵敏“微量免疫分析系统”的研制及其在癌症标志物测定中的应用

• 批准号: 17550088
• 负责人: TSUKAGOSHI Kazuhiko
• 金额: $ 1.98万
• 依托单位: Doshisha University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 2005
• 资助国家: 日本
• 起止时间: 2005 至 2006
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-17550088/
• 关键词: chemiluminescence; immune reaction; electrophoresis; micro-chip; protein; cancer marker; (7) (8); 免疫分析

We developed a chip analysis system incorporating electrophoresis-chemiluminescence detection, in which the chemiluminescence reaction of isoluminol isothiocyanato (ILITC) (chemiluminescence reagent for labeling)-microperoxidase (catalyst) - hydrogen peroxide (oxidant) as well as bis[2-(3,6,9-trioxadecanyloxycarbony)-4-nitrophenyl]oxalate (TDPO) (oxalate reagent)-fluorescein isothiocyanate (FITC) (fluorescence reagent for labeling) - hydrogen peroxide (oxidant) were adopted. The analysis system performed the following three processes on a chip having micro-channels of 20 μm deep and 50 μm width: immune reaction for high selectivity, electrophoresis for formation and transportation of the sample plug, and chemiluminescence detection for high sensitivity. The three processes were compactly integrated onto the chip. As the first process, the immune reaction was performed using an antibody-immobilized glass bead. The glass bead was placed in one of the reservoirs along with antigen (analyt … More e) and a known amount of ILITC- or FITC-labeled antigen to set up a competitive immune reaction. For electrophoresis, as the second process, the reactant after the immune reaction was fed electrophoretically into the intersection resulting in a sample plug. The sample plug was then moved into another reservoir containing hydrogen peroxide or TDPO-hydrogen peroxide mixture solution. At this point, chemiluminescence detection was performed as the third process: the labeled antigen mixed with the reagent to produce chemiluminescence. Chemiluminescence was detected with a photomultiplier tube located under the reservoir. The chip analysis system described here was capable of determining, with high selectivity and sensitivity, human serum albumin or immunosuppressive acidic protein (IAP) as a cancer marker in human serum. IAP was analyzed by immune reaction, electrophoresis, and chemiluminescence detection on the microchip. IAP was determined over the range of 1.0x10^<-7> - 5.0x10^<-6> M (with a detection limit of 1.0x10^<-7> M (S/N=3) (correlation coefficient, 0.999). One analysis was performed within 2 min. Less

我们开发了一种电泳-化学发光检测一体化的芯片分析系统，其中异硫氰酸异鲁米醇（ILITC）（化学发光标记试剂）-微过氧化物酶（催化剂）-过氧化氢（氧化剂）以及双[2-(3, 6,9-三氧癸氧基羰基)-4-硝基苯基]草酸酯 (TDPO)采用(草酸盐试剂)-异硫氰酸荧光素(FITC)(标记用荧光试剂)-过氧化氢(氧化剂)分析系统在具有20μm深、50μm宽的微通道的芯片上进行以下三个过程：免疫。用于高选择性的反应、用于样品塞的形成和运输的电泳以及用于高灵敏度的化学发光检测这三个过程被紧凑地集成到芯片上。过程中，使用固定有抗体的玻璃珠进行免疫反应，将玻璃珠与抗原（分析物……更多 e）和已知量的 ILITC 或 FITC 标记抗原一起放入其中一个容器中，以建立一种抗体。对于电泳，作为第二个过程，将免疫反应后的反应物通过电泳输送到交叉点，形成样品塞，然后将样品塞移入另一个含有过氧化氢或TDPO-氢气的容器中。此时，进行化学发光检测作为第三过程：用位于储液器下方的光电倍增管检测标记的抗原，产生化学发光。具有高选择性和灵敏度，通过免疫反应、电泳和化学发光分析人血清白蛋白或免疫抑制酸性蛋白（IAP）作为人血清中的癌症标志物。在微芯片上检测 IAP 的范围为 1.0x10^<-7> - 5.0x10^<-6> M（检测限为 1.0x10^<-7> M (S/N=3) (相关系数，0.999）在 2 分钟内完成一次分析。

项目成果

Competitive immunoassay using capillary electrophoresis with chemiluminescence detector

使用毛细管电泳和化学发光检测器进行竞争性免疫分析

• 发表时间: 2005
• 期刊: Bulletin of the Chemical Society of Japan 78
• 作者: Kazuhiko Tsukagoshi;et al.
• 通讯作者: et al.

Enhancing effect of phenylboronic acid compounds and their interactions with the diol groups of saccharides in a capillary electrophoresis-chemiluminescece detection system

毛细管电泳-化学发光检测系统中苯基硼酸化合物的增强作用及其与糖类二醇基团的相互作用

• 发表时间: 2007
• 期刊: Analytical Sciences 23
• 作者: Kazuhiko Tsukagoshi;et al.
• 通讯作者: et al.

Enhancing Effect of Phenylboronic Acid Compounds and Their Interactions with the Diol Groups of Saccharides in a Capillary Electrophoresis-Chemiluminescence Detection System

• DOI: 10.2116/analsci.23.227
• 发表时间: 2007-02
• 期刊: Analytical Sciences
• 影响因子: 1.6
• 作者: K. Tsukagoshi;Futoshi Ueno;R. Nakajima;K. Araki

Capillary Electrophoresis with a Chemiluminescence Detector Using Both Luminol an Peroxyoxalate Systems

使用鲁米诺和过氧草酸盐系统的化学发光检测器进行毛细管电泳

• 期刊: Instrumentation Science & Technology (in press)
• 作者: Kazuhiko Tsukagoshi;et al.;Kazuhiko Tsukagoshi et al.;Kazuhiko Tsukagoshi et al.
• 通讯作者: Kazuhiko Tsukagoshi et al.

Molecular recognition of mono- and disaccharides through interaction with p-iodiphenylboronic acid in capillary electrophoresis with a chemiluminescence detection system

通过化学发光检测系统在毛细管电泳中与对碘二苯硼酸相互作用进行单糖和二糖的分子识别

• 发表时间: 2006
• 期刊: Journal of Chromatography A 1123
• 作者: Kazuhiko Tsukagoshi;et al.
• 通讯作者: et al.