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Expression of rhEPO-L-Fc fusion protein and analysis of its bioactivity and pharmacokinetics / 生物工程学报
Chinese Journal of Biotechnology ; (12): 1874-1879, 2008.
Artículo en Chino | WPRIM | ID: wpr-302898
ABSTRACT
To prolong serum half-life of human Erythropoietin for better efficacy, a new form of recombinant human erythropoietin (rhEpo-L-Fc) was generated by fusion of a full length human erythropoietin gene and the Fc fragment of human IgG1 with flexible linker sequence. The fusion gene rhEPO-L-Fc was constructed by PCR, then inserted into expression vector pOptiVEC-TOPO, and expressed in Chinese Hamster Ovary cells deficient in the DHFR enzyme(CHO-dhfr-). The chimeric protein was purified by Protein A affinity chromatography, showed expected molecular weight and demonstrated a similar bioactivity compared to that of the native recombinant human erythropoietin (rhEPO) in an EPO-dependent cell-based assay. In vivo pharmacokinetic studies showed that the rhEPO-L-Fc had an elimination half-life of 27 h. In vivo efficacy studies showed that a single dose administration of rhEPO-L-Fc in rats increased the reticulocyte number in the peripheral blood significantly. These results demonstrated that the new engineered rhEPO-L-Fc may become alternative therapeutic approach to extend the half-time of rhEPO to treat anemia.
Asunto(s)
Texto completo: Disponible Índice: WPRIM (Pacífico Occidental) Asunto principal: Proteínas Recombinantes de Fusión / Proteínas Recombinantes / Fragmentos Fc de Inmunoglobulinas / Farmacocinética / Química / Cricetulus / Eritropoyetina / Células CHO / Ratas Sprague-Dawley / Genética Límite: Animales / Femenino / Humanos / Masculino Idioma: Chino Revista: Chinese Journal of Biotechnology Año: 2008 Tipo del documento: Artículo

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Texto completo: Disponible Índice: WPRIM (Pacífico Occidental) Asunto principal: Proteínas Recombinantes de Fusión / Proteínas Recombinantes / Fragmentos Fc de Inmunoglobulinas / Farmacocinética / Química / Cricetulus / Eritropoyetina / Células CHO / Ratas Sprague-Dawley / Genética Límite: Animales / Femenino / Humanos / Masculino Idioma: Chino Revista: Chinese Journal of Biotechnology Año: 2008 Tipo del documento: Artículo