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Dadun > Depósito Académico > Facultad de Farmacia > Departamento de Farmacia y Tecnología Farmacéutica > DA - Farmacia - Tecnología Farmacéutica - Artículos de revista >

A simple and efficient method for the production of human glycosylated glial cell line-derived neurotrophic factor using a Semliki Forest virus expression system
Authors: Ansorena, E. (Eduardo)
Casales, E. (Erkuden)
Aranda, A. (Alejandro)
Tamayo, E. (Esther)
Garbayo, E. (Elisa)
Smerdou, C. (Cristian)
Blanco-Prieto, M.J. (María José)
Aymerich, M.S. (María S.)
Keywords: GDNF
SFV expression system
Protein expression
Protein purification
BHK cells
Glycosylated recombinant protein
Issue Date: 2013
Publisher: Elsevier
Publisher version: http://dx.doi.org/10.1016/j.ijpharm.2012.04.071
ISSN: 0378-5173
Citation: Ansorena E, Casales E, Aranda A, Tamayo E, Garbayo E, Smerdou C, et al. A simple and efficient method for the production of human glycosylated glial cell line-derived neurotrophic factor using a Semliki Forest virus expression system. Int J Pharm 2013 JAN 2;440(1):19-26
Abstract
Human glial cell line-derived neurotrophic factor (hGDNF) is a very promising protein for the treatment of Parkinson's disease and other neurodegenerative disorders. The present work describes a quick and simple method to obtain a high amount of purified hGDNF using a mammalian cell-derived system. The method is based on the high expression level provided by a Semliki Forest virus vector and its ability to induce a strong shut-off of host-cell protein synthesis in mammalian cells. As a result, hGDNF is the only protein present in the supernatant and can be efficiently purified by a single chromatographic step. Using this system it was possible to eliminate other secreted proteins from the culture medium, like insulin-like growth factor-5, which are hard to remove using other hGDNF production methods. Purified hGDNF presents a complex glycosylation pattern typical of mammalian expression systems and is biologically active. This protocol could be extended to other secreted proteins and could be easily scaled up for industrial purposes.
Permanent link: http://hdl.handle.net/10171/23071
Appears in Collections:DA - CIMA - Neurociencias - Neuroanatomía ganglios basales - Artículos de Revista
DA - CIMA - Terapia génica y Hepatología - Vectores - Artículos de revista
DA - Ciencias - Bioquímica y Biología molecular - Artículos de Revista
DA - Farmacia - Tecnología Farmacéutica - Artículos de revista

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