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Protein Expr Purif ; 159: 69-74, 2019 07.
Artigo em Inglês | MEDLINE | ID: mdl-30917920

RESUMO

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.


Assuntos
Fator Estimulador de Colônias de Granulócitos e Macrófagos/química , Fator Estimulador de Colônias de Granulócitos e Macrófagos/genética , Proteínas Recombinantes/química , Proteínas Recombinantes/genética , Animais , Baculoviridae/genética , Sequência de Bases , Bombyx/genética , Extratos Celulares/química , Linhagem Celular , Cromatografia de Afinidade , Expressão Gênica , Glicosilação , Humanos , Concentração de Íons de Hidrogênio , Estabilidade Proteica , Viroses
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