Development of recombinant human granulocyte colony-stimulating factor (nartograstim) production process in Escherichia coli compatible with industrial scale and with no antibiotics in the culture medium (2021)
- Authors:
- USP affiliated authors: MAKIYAMA, EDSON NAOTO - FCF ; GARCIA, PRIMAVERA BORELLI - FCF ; BARAZZONE, GIOVANA CAPPIO - Interunidades em Biotecnologia ; HO, PAULO LEE - IQ ; GONÇALVES, VIVIANE MAIMONI - Interunidades em Biotecnologia
- Unidades: FCF; Interunidades em Biotecnologia; IQ
- DOI: 10.1007/s00253-020-11014-y
- Subjects: GRANULÓCITOS; ESCHERICHIA COLI
- Agências de fomento:
- Language: Inglês
- Imprenta:
- Source:
- Título: Applied Microbiology and Biotechnology
- ISSN: The granulocyte colony-stimulating factor (G-CSF) is a hematopoietic cytokine that has important clinical applications for treating neutropenia. Nartograstim is a recombinant variant of human G-CSF. Nartograstim has been produced in Escherichia coli as inclusion bodies (IB) and presents higher stability and biological activity than the wild type of human G-CSF because of its mutations. We developed a production process of nartograstim in a 10-L bioreactor using auto-induction or chemically defined medium. After cell lysis, centrifugation, IB washing, and IB solubilization, the following three refolding methods were evaluated: diafiltration, dialysis, and direct dilution in two refolding buffers.Western blot and SDS-PAGE confirmed the identity of 18.8-kDa bands as nartograstim in both cultures. The auto-induction medium produced 1.17 g/L and chemically defined medium produced 0.95 g/L. The dilution method yielded the highest percentage of refolding (99%). After refolding, many contaminant proteins precipitated during pH adjustment to 5.2, increasing purity from 50 to 78%. After applying the supernatant to cation exchange chromatography (CEC), nartograstim recovery was low and the purity was 87%. However, when the refolding solution was applied to anion exchange chromatography followed by CEC, 91%–98%purity and 2.2%recovery were obtained. The purification process described in this work can be used to obtain nartograstim with high purity, structural integrity, and the expected biological activity
- Volume/Número/Paginação/Ano: v. 105, p. 169–183, 2021
- Este periódico é de assinatura
- Este artigo NÃO é de acesso aberto
- Cor do Acesso Aberto: closed
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ABNT
EGUIA, Fara A. P et al. Development of recombinant human granulocyte colony-stimulating factor (nartograstim) production process in Escherichia coli compatible with industrial scale and with no antibiotics in the culture medium. Applied Microbiology and Biotechnology, v. 105, p. 169–183, 2021Tradução . . Disponível em: https://doi.org/10.1007/s00253-020-11014-y. Acesso em: 12 jun. 2025. -
APA
Eguia, F. A. P., Mascarelli, D. E., Carvalho, E. de, Rodríguez, G. R., Makiyama, E. N., Borelli, P., et al. (2021). Development of recombinant human granulocyte colony-stimulating factor (nartograstim) production process in Escherichia coli compatible with industrial scale and with no antibiotics in the culture medium. Applied Microbiology and Biotechnology, 105, 169–183. doi:10.1007/s00253-020-11014-y -
NLM
Eguia FAP, Mascarelli DE, Carvalho E de, Rodríguez GR, Makiyama EN, Borelli P, Lieberman C, Ho PL, Barazzone GC, Gonçalves VM. Development of recombinant human granulocyte colony-stimulating factor (nartograstim) production process in Escherichia coli compatible with industrial scale and with no antibiotics in the culture medium [Internet]. Applied Microbiology and Biotechnology. 2021 ; 105 169–183.[citado 2025 jun. 12 ] Available from: https://doi.org/10.1007/s00253-020-11014-y -
Vancouver
Eguia FAP, Mascarelli DE, Carvalho E de, Rodríguez GR, Makiyama EN, Borelli P, Lieberman C, Ho PL, Barazzone GC, Gonçalves VM. Development of recombinant human granulocyte colony-stimulating factor (nartograstim) production process in Escherichia coli compatible with industrial scale and with no antibiotics in the culture medium [Internet]. Applied Microbiology and Biotechnology. 2021 ; 105 169–183.[citado 2025 jun. 12 ] Available from: https://doi.org/10.1007/s00253-020-11014-y - Autophagy is required for hemopoietic stem cell survival in protein malnutrition
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Informações sobre o DOI: 10.1007/s00253-020-11014-y (Fonte: oaDOI API)
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