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

7 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

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Fluorescence
Cell-Penetrating Peptides
Antigens
Single-Chain Antibodies
Immunoglobulin Fragments
Molecules
Osteocalcin
Cell membranes
Protein C
Fluorescent Dyes
Nucleation
Fusion reactions
Cells
Membranes
Imaging techniques
Amino Acids
Antibodies
Proteins

ASJC Scopus subject areas

  • Analytical Chemistry

Cite this

Flashbody : A Next Generation Fluobody with Fluorescence Intensity Enhanced by Antigen Binding. / Wongso, Devina; Dong, Jinhua; Ueda, Hiroshi; Kitaguchi, Tetsuya.

In: Analytical Chemistry, Vol. 89, No. 12, 20.06.2017, p. 6719-6725.

Research output: Contribution to journalArticle

Wongso, Devina ; Dong, Jinhua ; Ueda, Hiroshi ; Kitaguchi, Tetsuya. / Flashbody : A Next Generation Fluobody with Fluorescence Intensity Enhanced by Antigen Binding. In: Analytical Chemistry. 2017 ; Vol. 89, No. 12. pp. 6719-6725.
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