Development of High thorough put immunoassay using microchip electrophoresis

使用微芯片电泳的高彻底性免疫分析的开发

基本信息

批准号：
16590464
负责人：
ARAKAWA Hidetoshi
金额：
$ 2.3万
依托单位：
Showa University
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (C)
财政年份：
2004
资助国家：
日本
起止时间：
2004 至 2006
项目状态：
已结题

项目摘要

In this study, the high-speed immunoassay using capillary electrophoresis and microchip electrophoresis was examined. First the method using the capillary electrophoresis was examined. The detection limit is 10pg and the reproducibility was 4.31 (CV%). Next, the generation of the steady electroosmotic flow in the microchip electrophoresis was examined. This enabled the application to the immunoassay, and the wall in the channel of the microchip made of the plastic was ionized by the coating method of the anionic detergent. As the result, it was possible to confirm the electroosmotic flow in the good reproduction. This method was applied to zone electrophoresis separation of the amino acid. And, the application of the immunoassay was more difficult than the sensitivity being inferior about 100 times than the capillary electrophoresis on the microchip. Therefore, it seemed to be necessary to use the more supersensitive detection system for development of the immunoassay, and next, it examined microchip electrophoresis by the bio-and chemiluminescence detection. Though as the result, it was preliminary, ATP detection in the micro M order became possible, and the possibility of the immunoassay by the luminescence detection was suggested. In addition, in this study, DNA was used as a marker of labeled antigen, and the charge and method by the intercalation fluorescence dye were also examined. As the result, the separation of the DNA labeled steroid became possible, and the application to the immunoassay was examined. From results of this study, the high-speed immunoassay by the capillary electrophoresis became possible, the immunoassay by the microchip electrophoresis could not be achieved. However, it was possible to obtain many basic results by this study. In the future, high speed microchip immunoassay which can be used in the clinical field seems to become possible by using aptamer assay using the nucleic acid in spite of antibody

在这项研究中，检查了使用毛细管电泳和微芯片电泳的高速免疫测定。首先，检查了使用毛细管电泳的方法。检测极限为10pg，可重复性为4.31（CV％）。接下来，检查了微芯片电泳中稳定的电流流的产生。这使得能够应用于免疫测定法，以及通过阴离子洗涤剂的涂层方法将由塑料制成的微芯片通道中的壁电离化。结果，有可能在良好的繁殖中确认电流流。该方法应用于氨基酸的区域电泳分离。而且，免疫测定的应用要比微芯片上的毛细管电泳的灵敏度低约100倍。因此，似乎有必要使用更超敏感的检测系统来开发免疫测定法，其次，它通过生物发光检测检查了微芯片电泳。尽管结果是初步的，但在微观序列中的ATP检测是可能的，并且提出了通过发光检测进行免疫测定的可能性。此外，在这项研究中，DNA被用作标记抗原的标记，还检查了插入荧光染料的电荷和方法。结果，可以进行标记为类固醇的DNA的分离，并检查了免疫测定的应用。从这项研究的结果来看，毛细管电泳的高速免疫测定是可能的，无法实现微芯片电泳的免疫测定。但是，通过这项研究可以获得许多基本结果。将来，可以在临床领域使用的高速微芯片免疫测定法似乎是通过使用核酸的适体分析来实现的

项目成果

期刊论文数量（22）

专著数量（0）

科研奖励数量（0）

会议论文数量（0）

专利数量（0）

Detection of cariogenic bacterial genes by microchip electrophoresis

微芯片电泳检测致龋细菌基因

DOI：
发表时间：
2004
期刊：
J. Chromatogr. B 810
影响因子：
0
作者：
K Higai;Y Aoki;Y Azuma;K Matsumoto;K.Karasawa
通讯作者：
K.Karasawa

Detection of cariogenic bacteria genes by a combination of allele-specific polymerase chain reactions and a novel bioluminescent pyrophosphate assay

DOI：
10.1016/j.ab.2004.06.026
发表时间：
2004-10-15
期刊：
ANALYTICAL BIOCHEMISTRY
影响因子：
2.9
作者：
Arakawa, H;Karasawa, K;Maeda, M
通讯作者：
Maeda, M

High sensitive and speedy DNA analysis using bioluminescence

使用生物发光进行高灵敏度、快速的 DNA 分析

DOI：
发表时间：
2004
期刊：
Pharmacia 41
影响因子：
0
作者：
Sato T;Yamamoto Y;Nakanishi T;Fukada K;Sugai F;Zhou Z;Okuno T;Nagano S;Hirata S;Shimizu A;Sakoda S;Azuma Yutaro;H.Arakawa
通讯作者：
H.Arakawa

The capillary electrophoresis separation of benzodiazepine drug using dextran sulfate and SDS as running buffer.

以硫酸葡聚糖和 SDS 作为电泳缓冲液，毛细管电泳分离苯二氮卓类药物。