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.
Artigo
em Chinês
| 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:
DisponíveL
Índice:
WPRIM (Pacífico Ocidental)
Assunto principal:
Proteínas Recombinantes de Fusão
/
Proteínas Recombinantes
/
Química
/
Cromatografia por Troca Iônica
/
Hirudinas
/
Ativador de Plasminogênio Tecidual
/
Dissulfetos
/
Redobramento de Proteína
/
Fibrinolíticos
/
Métodos
Idioma:
Chinês
Revista:
Chinese Journal of Biotechnology
Ano de publicação:
2010
Tipo de documento:
Artigo
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