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Biochem Biophys Res Commun. 2014 Feb 28;445(1):1-5. doi: 10.1016/j.bbrc.2013.12.102. Epub 2014 Jan 25.

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

Author information

1
Division of Biotechnology and Life Science, Institute of Engineering, Tokyo University of Agriculture and Technology, 2-24-16, Naka-cho, Koganei, Tokyo 184-8588, Japan.
2
Division of Biotechnology and Life Science, Institute of Engineering, Tokyo University of Agriculture and Technology, 2-24-16, Naka-cho, Koganei, Tokyo 184-8588, Japan. Electronic address: y-tomoko@cc.tuat.ac.jp.

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.

KEYWORDS:

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

PMID:
24472552
DOI:
10.1016/j.bbrc.2013.12.102
[Indexed for MEDLINE]

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