Functional expression of an scFv on bacterial magnetic particles by in vitro docking

Yasuhiro Sugamata, Tsuyoshi Tanaka, Tadashi Matsunaga, Tomoko Yoshino

Research output: Contribution to journalArticle

5 Citations (Scopus)

Abstract

A Gram-negative, magnetotactic bacterium, Magnetospirillum magneticum AMB-1 produces nano-sized magnetic particles (BacMPs) in the cytoplasm. Although various applications of genetically engineered BacMPs have been demonstrated, such as immunoassay, ligand-receptor interaction or cell separation, by expressing a target protein on BacMPs, it has been difficult to express disulfide-bonded proteins on BacMPs due to lack of disulfide-bond formation in the cytoplasm. Here, we propose a novel dual expression system, called in vitro docking, of a disulfide-bonded protein on BacMPs by directing an immunoglobulin Fc-fused target protein to the periplasm and its docking protein ZZ on BacMPs. By in vitro docking, an scFv-Fc fusion protein was functionally expressed on BacMPs in the dimeric or trimeric form. Our novel disulfide-bonded protein expression system on BacMPs will be useful for efficient screening of potential ligands or drugs, analyzing ligand-receptor interactions or as a magnetic carrier for affinity purification.

Original languageEnglish
Pages (from-to)1-5
Number of pages5
JournalBiochemical and Biophysical Research Communications
Volume445
Issue number1
DOIs
Publication statusPublished - 2014 Feb 28
Externally publishedYes

Fingerprint

Disulfides
Proteins
Ligands
Magnetospirillum
Cytoplasm
Immunoglobulin Fc Fragments
Periplasm
Cell Separation
In Vitro Techniques
Gram-Negative Bacteria
Immunoassay
Purification
Bacteria
Screening
Fusion reactions
Pharmaceutical Preparations

Keywords

  • Bacterial magnetic particles (BacMPs)
  • Immunoglobulin Fc
  • In vitro docking
  • Magnetospirillum magneticum AMB-1
  • Single chain variable fragment (scFv)

ASJC Scopus subject areas

  • Biochemistry
  • Biophysics
  • Cell Biology
  • Molecular Biology

Cite this

Functional expression of an scFv on bacterial magnetic particles by in vitro docking. / Sugamata, Yasuhiro; Tanaka, Tsuyoshi; Matsunaga, Tadashi; Yoshino, Tomoko.

In: Biochemical and Biophysical Research Communications, Vol. 445, No. 1, 28.02.2014, p. 1-5.

Research output: Contribution to journalArticle

Sugamata, Yasuhiro ; Tanaka, Tsuyoshi ; Matsunaga, Tadashi ; Yoshino, Tomoko. / Functional expression of an scFv on bacterial magnetic particles by in vitro docking. In: Biochemical and Biophysical Research Communications. 2014 ; Vol. 445, No. 1. pp. 1-5.
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