論文 - 詳細
| RRC ID | 42721 |
|---|---|
| 著者 | Mikami S, Kobayashi T, Yokoyama S, Imataka H. |
| タイトル | A hybridoma-based in vitro translation system that efficiently synthesizes glycoproteins. |
| ジャーナル | J Biotechnol |
| Abstract |
Since a large number of eukaryotic proteins are glycoproteins, an efficient and easily available cell-free system for the production of recombinant glycoproteins is needed. We have successfully developed an efficient cell-free translation system derived from a monoclonal antibody-producing hybridoma for this purpose. While extracts from HeLa cells were very inefficient for production of an N-glycosylated form of human immunodeficiency virus type-1 envelope protein 120 (gp120), the hybridoma extract was able to fully N-glycosylate gp120. During cell-free translation, eIF2alpha and eIF2alpha-kinases in the hybridoma extracts were observed to become phosphorylated due to the presence of essential supplements creatine phosphate and ATP. Addition of recombinant GADD34 and/or K3L to the extract efficiently lowered the phosphorylation of eIF2alpha, and thereby increased protein synthesis. By using this improved system, biologically active human choriogonadotropin (hCG), a glycoprotein hormone consisting of alpha and beta subunits was successfully synthesized. In conclusion, the hybridoma extract supplemented with GADD34/K3L should become a useful tool to produce recombinant glycoproteins. |
| 巻・号 | 127(1) |
| ページ | 65-78 |
| 公開日 | 2006-12-15 |
| DOI | 10.1016/j.jbiotec.2006.06.018 |
| PII | S0168-1656(06)00535-9 |
| PMID | 16889861 |
| MeSH | Cell-Free System Chorionic Gonadotropin / biosynthesis* Glycoproteins / metabolism Glycosylation Hybridomas / metabolism Protein Processing, Post-Translational / physiology* Recombinant Proteins / biosynthesis* |
| IF | 3.503 |
| 引用数 | 39 |
| WOS 分野 | BIOTECHNOLOGY & APPLIED MICROBIOLOGY |
| オルトメトリクス指標 |
オルトメトリクス指標項目
|
| 最多言及媒体 | Patent(IFI CLAIMS) |
| 各媒体での言及数の合計 | 16 |
| 過去6か月間でのオルトメトリクス指標の変動値 | 6.0 |
| リソース情報 | |
| ヒト・動物細胞 | HF10B4(RCB0708) |