Purification of bioactive glycosylated recombinant glial cell line-derived neurotrophic factor

Glial cell line-derived neurotrophic factor (GDNF) neuroprotective effect on dopaminergic neurons has been described in vitro and in vivo, turning up as a promising drug for the treatment of Parkinson's disease. Unglycosylated bacteria-obtained GDNF has already been successfully delivered for a...

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Bibliographic Details
Published in:International journal of pharmaceutics 2007-11, Vol.344 (1), p.9-15
Main Authors: Garbayo, E., Ansorena, E., Lanciego, J.L., Aymerich, M.S., Blanco-Prieto, M.J.
Format: Article
Language:English
Subjects:
Animals
Biological and medical sciences
Cell Line
Cells, Cultured
Cricetinae
GDNF
General pharmacology
Glial Cell Line-Derived Neurotrophic Factor - isolation & purification
Glial Cell Line-Derived Neurotrophic Factor - pharmacology
Glycosylation
Medical sciences
Neurites - drug effects
Neurites - physiology
Parkinson's disease
Pharmaceutical technology. Pharmaceutical industry
Pharmacology. Drug treatments
Protein purification
Rats
Recombinant Proteins - isolation & purification
Recombinant Proteins - pharmacology
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Summary:Glial cell line-derived neurotrophic factor (GDNF) neuroprotective effect on dopaminergic neurons has been described in vitro and in vivo, turning up as a promising drug for the treatment of Parkinson's disease. Unglycosylated bacteria-obtained GDNF has already been successfully delivered for a long period of time through an infusion pump directly to the putamen of Parkinsonian patients. Nevertheless, improved distribution and safety issues need to be solved and alternative strategies to long-term delivery seem necessary. The use of glycosylated GDNF could eliminate some safety concerns regarding the presence of antibodies against exogenous unglycosylated GDNF used for the treatment. Therefore, we have chosen a mammalian expression system as a source of glycosylated GDNF. In the present work, we describe the purification of recombinant rat GDNF from the culture media of baby hamster kidney (BHK) cells through several purification steps. Highly pure N-glycosylated recombinant GDNF has been obtained similar to the endogenous protein. Furthermore, the purified protein is biologically active when tested its ability to induce PC12 neurite outgrowth.
ISSN:0378-5173
1873-3476
DOI:10.1016/j.ijpharm.2007.04.003