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

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• 作者：Yasuhiro Sugamata ; Tsuyoshi Tanaka ; Tadashi Matsunaga ; Tomoko Yoshino ; ^{y-tomoko@cc.tuat.ac.jp" class="auth_mail}
• 关键词：In vitro docking ; Magnetospirillum magneticum AMB-1 ; Bacterial magnetic particles (BacMPs) ; Single chain variable fragment (scFv) ; Immunoglobulin Fc
• 刊名：Biochemical and Biophysical Research Communications
• 出版年：28 February, 2014
• 年：2014
• 卷：445
• 期：1
• 页码：1-5
• 全文大小：553 K

文摘

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.