Development of a novel bioluminescent probe and visualization measurement of biomolecules
| Project/Area Number |
16K21060
|
|---|
| Research Category |
Grant-in-Aid for Young Scientists (B)
|
| Allocation Type | Multi-year Fund |
|---|
| Research Field |
Chemical biology
Biomedical engineering/Biomaterial science and engineering
|
|---|
| Research Institution | Okinawa Institute of Science and Technology Graduate University (2018)
Japan Advanced Institute of Science and Technology (2016-2017) |
|---|
Principal Investigator |
Fukunaga Keisuke 沖縄科学技術大学院大学, 核酸化学・工学ユニット, 研究員 (80639279)
|
|---|
| Project Period (FY) |
2016-04-01 – 2019-03-31
|
| Project Status |
Completed (Fiscal Year 2018)
|
| Budget Amount *help |
¥4,030,000 (Direct Cost: ¥3,100,000、Indirect Cost: ¥930,000)
Fiscal Year 2017: ¥2,210,000 (Direct Cost: ¥1,700,000、Indirect Cost: ¥510,000)
Fiscal Year 2016: ¥1,820,000 (Direct Cost: ¥1,400,000、Indirect Cost: ¥420,000)
|
|---|
| Keywords | 生物発光共鳴エネルギー移動 / 抗体 / 蛍光 / 発光プローブ / 直交性反応 / 非天然アミノ酸 / 無細胞タンパク質合成系 / 人工タンパク質 / ルシフェラーゼ / クリック反応 / 発光 / 蛍光消光 / バイオ直交性反応 / 生物発光プローブ / バイオテクノロジー / 生体分子計測 / 免疫測定 |
|---|
| Outline of Final Research Achievements |
A novel bioluminescent resonance energy transfer (BRET) probe was synthesized by site-specific fluorescent labeling of luciferase-SNAP-single chain antibody (scFv) fusion with a SNAP-ligand. The developed BRET probe exhibited a different luminescent spectrum depending on the concentration of antigen. Also, site-selective modification method to functionalize N-terminal alpha amino-groups and N-glycans on monoclonal immunoglobulin G (IgG) antibody was established. On the ether hand, artificial luminescent protein that exhibits a red-shifted luminescence was generated by site-specific incorporation of fluorescent amino acid into luciferase.
|
| Academic Significance and Societal Importance of the Research Achievements |
位置選択的修飾技術を用いて抗体を蛍光標識することにより、抗体を蛍光・発光プローブへと変換する道筋が見えてきた。抗体を位置選択的に（多重）標識する技術は蛍光標識する以外にも用いることができるため、異なる機能を複数併せ持つ人工抗体の作製なども可能である。
|
Report
(4 results)
Research Products
(12 results)
