Expression and purification of biologically active human granulocyte-macrophage colony stimulating factor (hGM-CSF) using silkworm-baculovirus expression vector system.
Animals
Baculoviridae
/ genetics
Base Sequence
Bombyx
/ genetics
Cell Extracts
/ chemistry
Cell Line
Chromatography, Affinity
Gene Expression
Glycosylation
Granulocyte-Macrophage Colony-Stimulating Factor
/ chemistry
Humans
Hydrogen-Ion Concentration
Protein Stability
Recombinant Proteins
/ chemistry
Virus Diseases
Granulocyte-macrophage colony-stimulating factor
Recombinant proteins
Silkworm-baculovirus expression system
Journal
Protein expression and purification
ISSN: 1096-0279
Titre abrégé: Protein Expr Purif
Pays: United States
ID NLM: 9101496
Informations de publication
Date de publication:
07 2019
07 2019
Historique:
received:
12
02
2019
accepted:
19
03
2019
pubmed:
29
3
2019
medline:
28
4
2020
entrez:
29
3
2019
Statut:
ppublish
Résumé
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.
Identifiants
pubmed: 30917920
pii: S1046-5928(19)30082-8
doi: 10.1016/j.pep.2019.03.010
pii:
doi:
Substances chimiques
Cell Extracts
0
Recombinant Proteins
0
Granulocyte-Macrophage Colony-Stimulating Factor
83869-56-1
Types de publication
Journal Article
Research Support, Non-U.S. Gov't
Langues
eng
Sous-ensembles de citation
IM
Pagination
69-74Informations de copyright
Copyright © 2019 Elsevier Inc. All rights reserved.