Flashbody: A Next Generation Fluobody with Fluorescence Intensity Enhanced by Antigen Binding

Devina Wongso, Jinhua Dong, Hiroshi Ueda*, Tetsuya Kitaguchi

*この研究の対応する著者

研究成果: Article査読

23 被引用数 (Scopus)

抄録

Fluorescent probes are valuable tools for visualizing the spatiotemporal dynamics of molecules in living cells. Here we developed a genetically encoded antibody probe with antigen-dependent fluorescence intensity called "Flashbody". We first created a fusion of EGFP to the single chain variable region fragment (scFv) of antibody against seven amino acids of the bone Gla protein C-terminus (BGPC7) called BGP Fluobody, which successfully showed the intracellular localization of BGPC7-tagged protein. To generate BGP Flashbody, circularly permuted GFP was inserted in between two variable region fragments, and the linkers were optimized, resulting in fluorescence intensity increase of 300% upon binding with BGPC7 in a dose-dependent manner. Live-cell imaging using BGP Flashbody showed that BGPC7 fused with cell penetrating peptide was able to enter through the plasma membrane by forming a nucleation zone, while it penetrated the nuclear membrane with different mechanism. The construction of Flashbody will be possible for a range of antibody fragments and opens up new possibilities for visualizing a myriad of molecules of interest.

本文言語English
ページ(範囲)6719-6725
ページ数7
ジャーナルAnalytical Chemistry
89
12
DOI
出版ステータスPublished - 2017 6月 20

ASJC Scopus subject areas

  • 分析化学

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