Novel Expansion of Bioanalysis Based on the Fluorescence Quench-Release Principle

2014 Fiscal Year Final Research Report

• Project/Area Number: 24360336
• Research Category: Grant-in-Aid for Scientific Research (B)
• Allocation Type: Partial Multi-year Fund
• Section: 一般
• Research Field: Biofunction/Bioprocess
• Research Institution: Tokyo Institute of Technology
• Principal Investigator: UEDA Hiroshi 東京工業大学, 資源化学研究所, 教授 (60232758)
• Co-Investigator(Kenkyū-buntansha): DONG Jinhua 東京工業大学, 資源化学研究所, 助教 (80527838)
• Co-Investigator(Renkei-kenkyūsha): MATSUYAMA Yuki (OHMURO Yuki) 神戸大学, 大学院工学研究科, 特命助教 (30571088)
• Research Collaborator: ABE Ryoji ウシオ電機, 研究員
• Project Period (FY): 2012-04-01 – 2015-03-31
• Keywords: antibody / fluorescence quenching / immunoassay / cell-free translation / biochip / protein engineering / chemical biology / bioimaging

Outline of Final Research Achievements

We aimed at the elucidation of basic principle of Quenchbody (Q-body) and its application to various fields. Q-body is a site-specifically fluorescence-labeled recombinant antibody fragment, which was recently discovered by ourselves. It can detect various antigens with high sensitivity and rapidity in situ.
We succeeded in synthesizing Fab-type Q-bodies not only using cell-free translation system but also using cell-based system, and found novel suitable fluorescent dyes and novel modification sites on antibody. We also found a way to make genetically encoded Q-bodies using GFP variants, succeeded in a bead-based assay and imaging of osteocalcin peptide secreted from the cells, and in creating a similar biosensor protein based on the other ligand binders called Q'-body.

Free Research Field

タンパク質工学