Development of a novel bioluminescent probe and visualization measurement of biomolecules

2018 Fiscal Year Final Research Report

• 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
• 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

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.

Free Research Field

ケミカルバイオロジー

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。