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

在本研究中，对使用毛细管电泳和微芯片电泳的高速免疫测定进行了检验。首先检查使用毛细管电泳的方法。检测限为10pg，重现性为4.31（CV%）。接下来，检查了微芯片电泳中稳定电渗流的产生。这使得能够应用于免疫测定，并且通过阴离子清洁剂的涂覆方法使由塑料制成的微芯片的通道壁离子化。结果，可以确认良好再现中的电渗流。将该方法应用于氨基酸的区带电泳分离。并且，免疫测定的应用更加困难，其灵敏度比微芯片上的毛细管电泳差约100倍。因此，似乎有必要使用更超灵敏的检测系统来开发免疫分析，接下来，通过生物和化学发光检测来检查微芯片电泳。虽然其结果是初步的，但微M量级的ATP检测成为可能，并且暗示了通过发光检测进行免疫测定的可能性。此外，在本研究中，使用DNA作为标记抗原的标记物，并且还研究了嵌入荧光染料的电荷和方法。结果，DNA标记的类固醇的分离成为可能，并且检验了其在免疫测定中的应用。从本研究的结果来看，毛细管电泳的高速免疫测定成为可能，而微芯片电泳的免疫测定无法实现。然而，通过这项研究可以获得许多基本结果。未来，通过使用核酸（尽管有抗体）的适体测定，可用于临床领域的高速微芯片免疫测定似乎成为可能

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。