Chromatography-assisted refolding of a fusion protein containing multiple disulfide bonds / 生物工程学报
Chinese Journal of Biotechnology
;
(12): 1157-1164, 2010.
Artículo
en Chino
| WPRIM
| ID: wpr-292157
ABSTRACT
To establish a refolding process for the protein fused with 12-peptide of hirudin and reteplase (HV12p-rPA), we developed an anion-exchange chromatography assisted method to form correct disulfide bonds. After evaluating various parameters by orthogonal experiments with Q Sepharose XL as refolding medium, we found that urea gradient, sample loading size and L-Arg concentration were three major factors to affect the refolding outcomes, and urea gradient was critical to the recovery yield. Meanwhile, enzymatic activity of the refolded protein was decreased by the increase of sample loading size, and the optimal concentration of L-Arg in the eluting buffer was 1 mol/L. Thus, a dual-gradient of urea and pH on the anion-exchange chromatography resulted in remarkable increase of specific fibrinolytic and anti-coagulative activities of the refolded protein. Compared with the dilution method for refolding HV12p-rPA, the present approach was more effective and advantageous.
Texto completo:
Disponible
Índice:
WPRIM (Pacífico Occidental)
Asunto principal:
Proteínas Recombinantes de Fusión
/
Proteínas Recombinantes
/
Química
/
Cromatografía por Intercambio Iónico
/
Hirudinas
/
Activador de Tejido Plasminógeno
/
Disulfuros
/
Replegamiento Proteico
/
Fibrinolíticos
/
Métodos
Idioma:
Chino
Revista:
Chinese Journal of Biotechnology
Año:
2010
Tipo del documento:
Artículo
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