| 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
|
| リソース情報 |
| ヒト・動物細胞 |
HF10B4(RCB0708) |
