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

Devina Wongso, Jinhua Dong, Hiroshi Ueda, Tetsuya Kitaguchi

Research output: Contribution to journalArticle

14 Citations (Scopus)

Abstract

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.

Original languageEnglish
Pages (from-to)6719-6725
Number of pages7
JournalAnalytical Chemistry
Volume89
Issue number12
DOIs
Publication statusPublished - 2017 Jun 20

ASJC Scopus subject areas

  • Analytical Chemistry

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