RRC ID |
58992
|
著者 |
Kinoshita Y, Xu J, Masuda A, Minamihata K, Kamiya N, Mon H, Fujita R, Kusakabe T, Lee JM.
|
タイトル |
Expression and purification of biologically active human granulocyte-macrophage colony stimulating factor (hGM-CSF) using silkworm-baculovirus expression vector system.
|
ジャーナル |
Protein Expr Purif
|
Abstract |
Human granulocyte-macrophage colony stimulating factor (hGM-CSF) is a hematopoietic growth factor. It is widely employed as a therapeutic agent targeting neutropenia in cancer patients undergoing chemotherapy and in patients with AIDS or after bone marrow transplantation. In this study, we constructed the recombinant baculoviruses for the expression of recombinant hGM-CSF (rhGM-CSF) with two small affinity tags (His-tag and Strep-tag) at the N or C-terminus. Compared to N-tagged rhGM-CSF, C-tagged rhGM-CSF was highly recovered from silkworm hemolymph. The purified rhGM-CSF proteins migrated as a diffuse band and were confirmed to hold N-glycosylations. A comparable activity was achieved when commercial hGM-CSF was tested as a control. Considering the high price of hGM-CSF in the market, our results and strategies using silkworm-baculovirus system can become a great reference for mass production of the active rhGM-CSF at a lower cost.
|
巻・号 |
159
|
ページ |
69-74
|
公開日 |
2019-7-1
|
DOI |
10.1016/j.pep.2019.03.010
|
PII |
S1046-5928(19)30082-8
|
PMID |
30917920
|
MeSH |
Animals
Baculoviridae / genetics
Base Sequence
Bombyx / genetics
Cell Extracts / chemistry
Cell Line
Chromatography, Affinity
Gene Expression
Glycosylation
Granulocyte-Macrophage Colony-Stimulating Factor / chemistry*
Granulocyte-Macrophage Colony-Stimulating Factor / genetics*
Humans
Hydrogen-Ion Concentration
Protein Stability
Recombinant Proteins / chemistry*
Recombinant Proteins / genetics*
Virus Diseases
|
IF |
1.513
|
引用数 |
4
|
リソース情報 |
カイコ |
n17 |