論文 - 詳細
| RRC ID | 34900 |
|---|---|
| 著者 | Miura T, Nagamune T, Kawahara M. |
| タイトル | Ligand-inducible dimeric antibody for selecting antibodies against a membrane protein based on mammalian cell proliferation. |
| ジャーナル | Biotechnol Bioeng |
| Abstract |
A method for selecting antibodies against a membrane protein is important for attaining a variety of antibody-based diagnostics and therapies. In this study, we propose a novel system to select specific antibodies against a membrane protein based on mammalian cell proliferation as a readout. The system employs a chimeric membrane protein in which a target membrane protein of interest is fused to the intracellular signaling domain of a cytokine receptor. The chimeric membrane protein transduces a cell proliferation signal through dimerization when co-expressed with a specific single-chain Fv fused with a mutant of FK-binding protein 12 (scFv-Fk) that can be conditionally dimerized by a synthetic ligand AP20187. To demonstrate this system, ErbB2 and gp130 were chosen as the target membrane protein and cytokine receptor, respectively. Consequently, co-expression of the ErbB2/gp130 chimera and ErbB2-specific scFv-Fk rendered the cells proliferative in response to AP20187. The system also allowed selection of high-affinity binders from a mixture composed of dominant low-affinity binders. This system may be extended to affinity maturation of scFvs by modulating AP20187 concentration in the selection process. |
| 巻・号 | 113(5) |
| ページ | 1113-23 |
| 公開日 | 2016-5-1 |
| DOI | 10.1002/bit.25858 |
| PMID | 26479395 |
| MeSH | Animals Cell Line Cell Proliferation Cytokine Receptor gp130 / genetics* Humans Ligands Mutation Protein Engineering / methods* Receptor, ErbB-2 / genetics* Receptor, ErbB-2 / immunology Recombinant Fusion Proteins / genetics Single-Chain Antibodies / genetics* Single-Chain Antibodies / immunology Tacrolimus Binding Protein 1A / genetics* Transduction, Genetic |
| IF | 4.002 |
| 引用数 | 2 |
| WOS 分野 | BIOTECHNOLOGY & APPLIED MICROBIOLOGY |
| リソース情報 | |
| 遺伝子材料 | pSV2 erb B2 (RDB01273) |
| ヒト・動物細胞 | Ba/F3(RCB0805) |