RRC ID 58992
Author Kinoshita Y, Xu J, Masuda A, Minamihata K, Kamiya N, Mon H, Fujita R, Kusakabe T, Lee JM.
Title Expression and purification of biologically active human granulocyte-macrophage colony stimulating factor (hGM-CSF) using silkworm-baculovirus expression vector system.
Journal 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.
Volume 159
Pages 69-74
Published 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
Times Cited 4
Resource
Silkworms n17