Laboratory for Chemistry and Life Science, Institute of Innovative Research, Tokyo Institute of Technology


LAST UPDATE 2020/06/08

  • 研究者氏名
    Researcher Name

    上田宏 Hiroshi UEDA
    教授 Professor
  • 所属
    Professional Affiliation

    Laboratory for Chemistry and Life Science, Institute of Innovative Research, Tokyo Institute of Technology

    Division of Molecular Bioscience
  • 研究キーワード
    Research Keywords


    protein engineering
    enzyme engineering
Research Subject
Development of novel bioprocesses based on protein engineering methodologies

研究の背景 Background


In nature, many proteins exhibit high performances such as molecular recognition ability and catalytic activity, which are still difficult to emulate artificially. However, natural protein does not always have ideal property from the viewpoint of application to the continuous development of human society. That is why we aim at creating novel proteins with superior performances and the assays using them, through the rational design and molecular evolution, as well as chemical biological techniques.

研究の目標 Outcome


Our primary target is the development and application of novel detection probes and assays based on recombinant antibodies, such as ‘Open sandwich immunoassay’ which can detect small molecules easily and sensitively, as well as ‘Quenchbodies’ which can detect various antigens by the increase of fluorescence. Secondly, we aims at constructing various novel processes based on enzyme engineering, such as development of novel protein-protein interaction assay FlimPIA, based on firefly luciferase mutants.

研究図Research Figure

Fig.1. Principle of Open-sandwich Immunoassay (Upper left). The two antigen-binding fragments VH and VL associate stronger in the presence of antigen, whose association is detected by the immobilized enzyme activityFig. 2. Working model of Quenchbody (Lower left). When the antibody VH and/or VL fragments are N-terminally labeled with fluorescent dye, it is quenched by the Trp residues in the antibody. Upon adding antigen, the V region is stabilized and the dye no longer interacts with Trp, resulting in increased fluorescence. Fig. 3. Principle of FlimPIA. Fluc mutant whose oxidative luminescent reaction is slow (donor) and another mutant whose adenylation reaction is slow (acceptor) are used. When the donor and acceptor come close at the time of interaction, luminescence activity is enhanced by the relay of reaction intermediate LH2-AMP.

文献 / Publications

ACS Sens. 5, 24 (2020). Biotechnol. Bioeng. 117, 1259 (2020). Sci. Rep. 9, 18189 (2019). Sci. Rep. 9, 10188 (2019). Biochemistry 58, 2695 (2019). Anal. Chem. 90, 3001 (2018). Chem. Commun. 53, 10200 (2017). WIREs Nanomed. Nanobiotechnol., e1457 (2017).