| Project/Area Number |
02650541
|
|---|
| Research Category |
Grant-in-Aid for General Scientific Research (C)
|
| Allocation Type | Single-year Grants |
|---|
| Research Field |
工業分析化学
|
|---|
| Research Institution | KYOTO UNIVERSITY |
|---|
Principal Investigator |
MORISHITA Fujio KYOTO UNIVERSITY Graduate School of Engineering, Associate Professor, 工学研究科, 助教授 (30026281)
|
|---|
| Project Period (FY) |
1990 – 1991
|
| Project Status |
Completed (Fiscal Year 1991)
|
| Budget Amount *help |
¥1,800,000 (Direct Cost: ¥1,800,000)
Fiscal Year 1991: ¥700,000 (Direct Cost: ¥700,000)
Fiscal Year 1990: ¥1,100,000 (Direct Cost: ¥1,100,000)
|
|---|
| Keywords | Flow injection analysis / Latex / Immobilized enzyme / Liposome / Immunoassay / Immobilized antigen / Immobilized antibody / Flow injection liposome immunoassay / フロ-インジェクション分析 / リポソ-ム / 免疫反応 / 抗体固定化反応管 / 抗原修飾リポソ-ム / フロ-インジェクション分析(FIA) / 酵素固定化ラテックス / リンゴ酸脱水素酵素 / NADH / 酵素的サイクリング / 酵素固定化中空キャピラリ-反応管 / 反応速度の比較 |
|---|
| Research Abstract |
A New Flow injection method (FIA) was proposed in order to improve the reaction efficiency, in which enzyme was immobilized on the surface of synthetic polymer latex. Stirene-acrylic acid copolymer latex synthesized was spherical and had homogeneous size of about 0.5 mm diameter. The enzymatic activity of enzyme-immobilized latex was dependent on the molar fraction of acrylic acid. Lactate dehydrogenase immobilized on the latex particles kept the activity higher than 50% of original one for two months or longer. The results obtained in FIA using latex suspended in the carrier solution were comparable to those in the conventional FIA using enzyme-immobilized reactors. Difficulties in recovering and reusing the used latex were the serious problem.
Another new FIA method (flow injection liposome immunoassay : FILIA) was discussed, in which antigen-immobilized liposome having marker molecules in its inner space was used and a competitive immunoassay was carried out in a continuous flow system. The sufficient activity of antibody immobilized on the wall of a reactor was confirmed by observing satisfactory results in a flow injection enzyme immunoassay using POD-conjugated antigen. However, any explicit proof for the immunoreaction could not be found in FILIA because of steric hindrance by bulky liposome and wide wall of the reactor. By inserting a hydrophillic polymer spacer between antigen and liposome or between antibody and the reactor wall, a sigmoid response was obtained for the free antigen injected as a sample. But the response was insufficient for the determination of low concentration of the free analyte.
|
