2019 Fiscal Year Final Research Report
Depelopment of immunochemical fluorescent sensor on the basis of tripartite split-GFP
| Project/Area Number |
17K15466
|
|---|
| Research Category |
Grant-in-Aid for Young Scientists (B)
|
| Allocation Type | Multi-year Fund |
|---|
| Research Field |
Natural medicines
|
|---|
| Research Institution | Kyushu University |
|---|
Principal Investigator |
|
|---|
| Project Period (FY) |
2017-04-01 – 2020-03-31
|
| Keywords | 小型化抗体 / ハリントニン / ELISA |
|---|
| Outline of Final Research Achievements |
This study aimed to develop rapid and sensitive immunological fluorescent sensor that can determine harringtonine (HT).
Primarily, VH-GFP10, GFP11-VL, and GFP1-9 genes were constructed to detect fluorescence in the presence of HT. However, the fluorescence derived from quaternary complex was not detected using this system. Subsequently, VH-GFP10-GFP11-VL (scFvGs) gene was constructed, in which VH-GFP10 and GFP11-VL was joined via flexible linker peptides of (GGGGS)n(n=2, 3). When the activity of scFvGs was investigated by using ELISA, it showed specific competitive activity to HT. Therefore, scFvGs were applied to immunological fluorescent sensor to detect fluorescence. As a result, slight fluorescence was detected compared with control. However, it was too weak to apply to quantitative analysis, suggesting that more optimization is required for development of this system.
|
| Free Research Field |
分析化学
|
| Academic Significance and Societal Importance of the Research Achievements |
本研究における対象化合物は低分子化合物であることから、二つの波及効果が考えられる。
一つは、違法薬物問題への貢献である。近年、社会問題となっている違法薬物は、その大部分が低分子化合物である。そのため、対象化合物HTを危険ドラッグや麻薬関連化合物へ置き換える事で社会問題となっている違法薬物の簡易迅速検出法へと応用できる。もう一つは、医薬品シーズ探索研究への貢献である。本システムの抗体遺伝子を他の受容体や疾病関連タンパク質に置き換えることで、無限の可能性を有する薬用資源から網羅的なスクリーニングが行なえ医薬品シーズの探索が可能となる。
本研究は、違法薬物問題や希少疾病医薬品開発問題の解決にも繋がる。
|
