Optimization of the primary recovery of human interferon alpha2b from Escherichia coli inclusion bodies.
Protein Expr Purif
; 45(1): 226-34, 2006 Jan.
Article
en En
| MEDLINE
| ID: mdl-16139511
ABSTRACT
The human interferon alpha2b (hu-IFNalpha2b) gene was cloned in Escherichia coli JM109(DE3) and the recombinant protein was expressed as cytoplasmic inclusion bodies (IB). The present work discusses the recovery of hu-IFNalpha2b IB from the E. coli cells. An optimized protocol is proposed based on the sequential evaluation of recovery steps and parameters (i) cell disruption, (ii) IB recovery and separation from cell debris, (iii) IB washing, and (iv) IB solubilization. Parameters such as hu-IFNalpha2b purity and recovery yield were measured after each step. The optimized recovery protocol yielded 60% of hu-IFNalpha2b with a purity of up to 80%. The protein was renatured at high concentration after recovery and it was found to display biological activity.
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Colección:
01-internacional
Base de datos:
MEDLINE
Asunto principal:
Cuerpos de Inclusión
/
Interferón-alfa
/
Escherichia coli
Límite:
Animals
/
Humans
Idioma:
En
Revista:
Protein Expr Purif
Asunto de la revista:
BIOLOGIA MOLECULAR
Año:
2006
Tipo del documento:
Article
País de afiliación:
Portugal