| Autor: |
Zeinab, Mohammadi, Mahdi, Alijanianzadeh, Rassoul, Khalilzadeh, Sirus, Khodadadi |
| Rok vydání: |
2022 |
| Předmět: |
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| Zdroj: |
Protein & Peptide Letters. 29:293-305 |
| ISSN: |
0929-8665 |
| DOI: |
10.2174/0929866529666220126100559 |
| Popis: |
Background and objective: Recombinant human granulocyte-colony stimulating factor (rhG-CSF) and its PEGylated form (PEG-GCSF) are used in the cancer therapy. Thus the development of a more cost-effectively method for expressing rhG-CSF and the PEGylation optimization of rhG-CSF by reaction engineering and subsequent the purification strategy is necessary. Methods: RhG-CSF expression in Escherichia coli BL21 (DE3) was carried out by auto-induction batch fermentation and improved for maximizing rhG-CSF productivity. After that, purified rhG-CSF was PEGylated using methoxy polyethylene glycol propionaldehydes (mPEG20-ALD). The various conditions effect of extraction and purification of rhG-CSF and PEG-GCSF were assayed. Results: The assessment results revealed that auto-induction batch cultivation strategy had maximum productivity and rhG-CSF purity was more than 99%. The obtained Data of rhG-CSF PEGylation displayed that the optimized conditions of rhG-CSF PEGylation and purification enhanced hemogenisity PEG-GCSF and managed reaction toward optimal yield of PEG-GCSF (70%) and purity of 99.9%. Findings from FTIR, CD, and fluorescence spectroscopy and bioassay revealed that PEGylation was executed exactly in the rhG-CSF N-terminus, and products maintained their conformation properties. Conclusion: Overall, the developed approach expanded strategies for high yield rhG-CSF by simplified auto-induction batch fermentation system and rhG-CSF PEGylation, which are simple and time-saving, economical and high efficiency. |
| Databáze: |
OpenAIRE |
| Externí odkaz: |
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