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

我们开发了一个编码电泳 - 化学发光检测的芯片分析系统，其中伊斯蒂氨醇异硫氰酸盐（ILITC）（用于标记的化学发光试剂）的化学发光反应 - 微氧化酶（催化剂） - 氧化物（氧化剂）以及氧化物（氧化剂）（氧化）（氧化剂） BIS [2-（3,6,9-三氧化烷氧甲氯）-4-硝基苯基]草酸盐（TDPO）（Oxalate试剂） - 荧光素异硫氰酸酯（FITC）（FITC）（标记的荧光试剂） - 氧化物 - 氧化物（氧化）（氧化）。分析系统对具有20μm深度和50μm宽度的微通道的芯片进行了以下三个过程：高选择性的免疫反应，用于样品塞形成和运输的电泳以及化学发光检测的高敏感性。将三个过程紧凑到芯片上。在第一个过程中，使用抗体击倒玻璃珠进行免疫反应。将玻璃珠与抗原（分析员…更多）一起放置在其中一个储层中，以及已知的ILITC或FITC标记的抗原，以建立竞争性免疫反应。对于电泳，作为第二个过程，将免疫反应后的反应物电泳送入交叉点，导致样品塞。然后将样品塞移入另一个含有过氧化氢或TDPO氢过氧化氢混合溶液的储层。这一点，将化学发光检测作为第三个过程：与试剂混合的标记的抗原以产生化学发光。用位于储层下方的光电倍增管检测到化学发光。此处描述的芯片分析系统能够以高选择性和敏感性，人血清白蛋白或免疫抑制性酸性蛋白（IAP）作为人血清中的癌症标志物来确定。通过在微芯片上通过免疫反应，电泳和化学发光检测来分析IAP。在1.0x10^<-7> -5.0x10^<-6> m的范围内确定IAP（检测极限为1.0x10^<-7> m（s/n = 3）（相关系数，0.999）。在2分钟内进行了一个分析。

项目成果

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.

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.

Development of a micro total analysis system incorporating chemiluminescence detection and application to detection of cancer markers

• DOI: 10.1021/ac040133t
• 发表时间: 2005-03-15
• 期刊: ANALYTICAL CHEMISTRY
• 影响因子: 7.4
• 作者: Tsukagoshi, K;Jinno, N;Nakajima, R
• 通讯作者: Nakajima, R

Compact polytetrafluoroethylene assembly-type capillary electrophoresis with chemiluminescence detection.

• DOI: 10.1016/j.chroma.2006.06.101
• 发表时间: 2006-08
• 期刊: Journal of chromatography. A
• 作者: K. Tsukagoshi;Shingo Ishida;Yuichi Oda;K. Noda;R. Nakajima

Capillary electrophoresis apparatus equipped with a bioluminescence detector using a batch- or flow-type detection cell.

• DOI: 10.1016/j.chroma.2005.09.048
• 发表时间: 2005-11
• 期刊: Journal of chromatography. A
• 作者: K. Tsukagoshi;Masayuki Tahira;R. Nakajima