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Title: | 抗体のN末端特異的蛍光標識による新規抗原検出法の開発 |
Other Titles: | N-Terminally fluorescent-labeled antibodies that show fluorescence change upon antigen-binding |
Authors: | 芳坂, 貴弘 |
Authors(alternative): | Hohsaka, Takahiro |
Keywords: | 抗体 イムノアッセイ 蛍光 |
Issue Date: | 5-Jun-2018 |
Abstract: | 本研究では、既存のIgG抗体のN末端を特異的に蛍光標識することで、蛍光応答性抗体を作製する新規手法を開発した。弱酸性条件下で蛍光標識アルデヒドをIgG抗体と反応させることで、N末端特異的に蛍光基が付加され、得られたN末端蛍光標識抗体は、抗原の結合に伴って蛍光強度変化を示すことを明らかにした。また、FRETと蛍光消光解消を組み合わせることで、抗原を蛍光強度比の変化として検出可能な二重標識蛍光応答性抗体を合成すること、および、タグ化タンパク質の無細胞翻訳系での発現をリアルタイム検出することも可能であった。:In this study, a novel method was developed for producing N-terminally fluorescent-labeled IgG antibodies that show antigen-dependent fluorescence change. By reacting fluorescent-labeled aldehyde with IgG antibody under weakly acidic condition, the fluorescent group was specifically attached at the N terminus, and the resulting N-terminally fluorescent-labeled antibody showed fluorescence intensity changed upon the antigen binding. In addition, it is possible to synthesize dual-labeled fluorescence-responsive antibody that can detect the antigen-binding as a change in fluorescence intensity ratio, and to detect tagged proteins expressed in a cell-free translation system in a real-time manner. |
Description: | 挑戦的萌芽研究 研究期間:2015~2017 課題番号:15K13739 研究者番号:30263619 研究分野:生体関連化学 |
Language: | jpn |
URI: | http://hdl.handle.net/10119/15399 |
Appears in Collections: | 2017年度 (FY 2017)
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15K13739seika.pdf | | 472Kb | Adobe PDF | View/Open |
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