| Project/Area Number |
12555238
|
|---|
| Research Category |
Grant-in-Aid for Scientific Research (B)
|
| Allocation Type | Single-year Grants |
|---|
| Section | 展開研究 |
|---|
| Research Field |
工業分析化学
|
|---|
| Research Institution | Ariake National College of Technology |
|---|
Principal Investigator |
MASADOME Takashi Department of Chemical Science and Engineering Ariake National College of Technology Associate Professor, 物質工学科, 助教授 (30190341)
|
|---|
| Co-Investigator(Kenkyū-buntansha) |
TABEI Hisao Material Technology Dept Device Development Dept , NTT Advanced Technology Corp. General Manager, 先端基盤技術事業本部・デバイス技術部材料部門, 部門長
IMATO Toshihiko Graduate School of Engineering, Kyushu University Professor, 大学院・工学研究院・化学システム工学専攻, 教授 (50117066)
ASANO Yasukazu Hachinohe National College of Technology, 物質工学科, 教授 (80311108)
KUROKAWA Yoichi Department of Environmental Analysis Fukuoka Prefectural Health and Environment Research Institute Senior Researcher, 管理部・環境計測課, 専門研究員
|
|---|
| Project Period (FY) |
2000 – 2002
|
| Project Status |
Completed (Fiscal Year 2002)
|
| Budget Amount *help |
¥11,000,000 (Direct Cost: ¥11,000,000)
Fiscal Year 2002: ¥2,100,000 (Direct Cost: ¥2,100,000)
Fiscal Year 2001: ¥3,000,000 (Direct Cost: ¥3,000,000)
Fiscal Year 2000: ¥5,900,000 (Direct Cost: ¥5,900,000)
|
|---|
| Keywords | endocrine disruptors / Sequential injection / solid phase extracton / Antigen / antibody reaction / Beads suspension / protein A / マイクロビーズ / 酵素免疫法 / 抗原抗体反応 |
|---|
| Research Abstract |
A sequential injection analysis (SIA) technique, in which antibody-immobilized microbeads were transferred to a jet ring (JR) cell, was used in determination of carp vitellogenin (Vg). The determination is based on a sandwich immunoassay in which two types of reactions between anti carp Vg antibodies and carp Vg are used. Namely, the antibody for the first reaction step was immobilized on microbeads (Sephadex beads), and an antibody labeled with a horseradish peroxidase (HRP) was used in the second step of the reaction. A mixed solution of hydrogen peroxide and o-phenylenediamine (OPD) was used as the source of the chromophore in the reaction. The microbeads-immobilized antibody, Vg analyte, HRP-labeled antibody and the color developing solution were introduced automatically into the JR cell of the SIA system in a programmed sequence, and the absorbance of the oxidized OPD product was used to determine the amount of Vg present. The optimal incubation times for the immunoreaction for the first and the second steps were determined at 120 and 60 min, respectively, taking into account the sensitivity to the Vg determination. Under these conditions, a good linear correlation was obtained between Vg concentration and the absorbance of the oxidized OPD. The lower detection limit for the determination of Vg was about 5 ng ml 1 in this system. The method developed here represents a simple, accurate method for the determination method of Vg.
|
