Reference - Detail
| RRC ID | 42721 |
|---|---|
| Author | Mikami S, Kobayashi T, Yokoyama S, Imataka H. |
| Title | A hybridoma-based in vitro translation system that efficiently synthesizes glycoproteins. |
| Journal | 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. |
| Volume | 127(1) |
| Pages | 65-78 |
| Published | 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 |
| Times Cited | 39 |
| WOS Category | BIOTECHNOLOGY & APPLIED MICROBIOLOGY |
| Altmetric score |
オルトメトリクス指標項目
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| The most frequently cited source | Patent(IFI CLAIMS) |
| Total number of mentions | 16 |
| Altmetric score changes over past 6months | 6.0 |
| Resource | |
| Human and Animal Cells | HF10B4(RCB0708) |