权益分类	功能权益	普通用户	{{item.name}}会员
{{category.name}}	{{benefitItem.name}}

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

本研究建立了毛细管电泳法和芯片电泳法相结合的高速免疫分析方法。首先对毛细管电泳法进行了考察。方法检出限为10pg，重现性为4.31(CV%)。其次，研究了微芯片电泳法中稳定电渗流的产生。这使得将其应用于免疫分析成为可能，由塑料制成的微芯片通道内的壁用阴离子洗涤剂的涂覆方法进行了电离。因此，有可能证实良好繁殖过程中的电渗流。将该方法用于氨基酸的区带电泳法分离。而且，免疫分析的应用难度大于灵敏度，比芯片上的毛细管电泳差100倍左右。因此，有必要使用更高灵敏度的检测系统来开发免疫分析方法，然后通过生物和化学发光检测来检测芯片电泳。虽然结果是初步的，但使微米量级的ATP检测成为可能，并提出了利用发光检测进行免疫分析的可能性。此外，本研究还以DNA作为标记抗原的标记物，并对嵌入荧光染料的电荷和方法进行了考察。从而使DNA标记的类固醇的分离成为可能，并对其在免疫分析中的应用进行了研究。从本研究的结果来看，毛细管电泳法的高速免疫分析成为可能，而微芯片电泳法的免疫分析无法实现。然而，通过这项研究有可能获得许多基本结果。在未来，利用核酸而不考虑抗体的核酸适体分析技术，使应用于临床领域的高速微芯片免疫分析成为可能。

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。