Production of highly pure human glycosylated GDNF in a mammalian cell line.
The administration of glial cell line-derived neurotrophic factor (GDNF) has emerged as a promising strategy for the treatment of several diseases of the nervous system as Parkinson's disease, amyotrophic lateral sclerosis, spinal cord injury and nerve regeneration as well as ocular diseases and drug addictions. A procedure for the purification of human recombinant glycosylated GDNF using a mammalian expression system as the source of the protein is discussed in the present paper. The neurotrophic factor was purified using cation exchange chromatography and gel filtration. A human cell line was chosen as the source of therapeutic protein, since a recombinant protein with a structure and glycosylation pattern equivalent to the native form is desirable for its prospective therapeutic utilization. The activity of the highly pure protein obtained was confirmed with a cell-based bioassay. The purified protein is suitable for its in vivo evaluation in animals and for possible subsequent clinical application.
Ansorena
Eduardo
E
Department of Pharmacy and Pharmaceutical Technology, School of Pharmacy, University of Navarra, C/Irunlarrea 1, E-31080 Pamplona, Spain.
Garbayo
Elisa
E
Lanciego
José L
JL
Aymerich
María S
MS
Blanco-Prieto
María J
MJ
D000818:Animals; D001681:Biological Assay; D001709:Biotechnology / Q000379:methods*; D002411:Cation Exchange Resins; D002454:Cell Differentiation / Q000187:drug effects; D002850:Chromatography, Gel; D002852:Chromatography, Ion Exchange; D005347:Fibroblasts / Q000378:metabolism*; D051100:Glial Cell Line-Derived Neurotrophic Factor / Q000096:biosynthesis* / Q000235:genetics / Q000302:isolation & purification / Q000494:pharmacology; D006031:Glycosylation; D006801:Humans; D009474:Neurons / Q000187:drug effects / Q000473:pathology; D016716:PC12 Cells; D010641:Phenotype; D011499:Protein Processing, Post-Translational; D051381:Rats; D011994:Recombinant Proteins / Q000096:biosynthesis; D021241:Spectrometry, Mass, Electrospray Ionization; D053719:Tandem Mass Spectrometry; D014161:Transduction, Genetic
D002411:Cation Exchange Resins; C495039:GDNF protein, human; D051100:Glial Cell Line-Derived Neurotrophic Factor; D011994:Recombinant Proteins