Functional expression of an scFv on bacterial magnetic particles by in vitro docking.
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.
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.
Yasuhiro
Y
Sugamata
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.
Tsuyoshi
T
Tanaka
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.
Tadashi
T
Matsunaga
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.
Tomoko
T
Yoshino
D015153:Blotting, Western; D004220:Disulfides / Q000378:metabolism; D052203:Ferrosoferric Oxide / Q000378:metabolism*; D055944:Magnetosomes / Q000378:metabolism*; D042201:Magnetospirillum / Q000235:genetics / Q000378:metabolism*; D053758:Nanoparticles / Q000378:metabolism; D011485:Protein Binding; D011994:Recombinant Proteins / Q000378:metabolism; D057127:Single-Chain Antibodies / Q000235:genetics / Q000378:metabolism*
D004220:Disulfides; D011994:Recombinant Proteins; D057127:Single-Chain Antibodies; D052203:Ferrosoferric Oxide
Bacterial magnetic particles (BacMPs); Immunoglobulin Fc; In vitro docking; Magnetospirillum magneticum AMB-1; Single chain variable fragment (scFv)