Extracellular production of recombinant N-glycosylated anti-VEGFR2 monobody in leaky Escherichia coli strain.
Biotechnol Lett
; 41(11): 1265-1274, 2019 Nov.
Article
en En
| MEDLINE
| ID: mdl-31541332
ABSTRACT
OBJECTIVE:
To improve the production yield of N-glycosylated anti-VEGFR2 (vascular endothelial growth factor receptor 2) monobody (FN3VEGFR2-Gly) in lpp knockout Escherichia coli cells harboring Campylobacter jejuni N-glycosylation pathway.RESULTS:
The leaky CLM37-Δlpp strain efficiently secreted FN3VEGFR2-Gly into culture medium. The extracellular levels of glycosylated FN3VEGFR2-Gly in CLM37-Δlpp culture medium were approximately 11 and 15 times higher compared to those in CLM37 cells via IPTG and auto-induction, respectively. In addition, the highest level of total glycosylated FN3VEGFR2-Gly (70 ± 3.4 mg/L) was found in culture medium via auto-induction. Furthermore, glycosylated FN3VEGFR2-Gly was more stable than unglycosylated FN3VEGFR2-Gly in this expression system, but their bioactivities were relatively similar.CONCLUSIONS:
Lpp knockout leaky E. coli strain combined with auto-induction method can enhance the extracellular production of homogenous N-glycosylated FN3VEGFR2-Gly, and facilitate the downstream protein purification. The findings of this study may provide practical implications for the large-scale production and cost-effective harvesting of N-glycosylation proteins.Palabras clave
Texto completo:
1
Colección:
01-internacional
Base de datos:
MEDLINE
Asunto principal:
Receptor 2 de Factores de Crecimiento Endotelial Vascular
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Escherichia coli
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Espacio Extracelular
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Dominio de Fibronectina del Tipo III
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Anticuerpos
Idioma:
En
Revista:
Biotechnol Lett
Año:
2019
Tipo del documento:
Article
País de afiliación:
China