Full metadata record
DC Field | Value | Language |
---|---|---|
dc.creator | Torres-Ortega, P.V. (Pablo Vicente) | - |
dc.creator | Smerdou, C. (Cristian) | - |
dc.creator | Ansorena-Artieda, E. (Eduardo) | - |
dc.creator | Ballesteros-Briones, M.C. (María Cristina) | - |
dc.creator | Martisova, E. (Eva) | - |
dc.creator | Garbayo, E. (Elisa) | - |
dc.creator | Blanco-Prieto, M.J. (María José) | - |
dc.date.accessioned | 2022-06-28T08:36:56Z | - |
dc.date.available | 2022-06-28T08:36:56Z | - |
dc.date.issued | 2021 | - |
dc.identifier.citation | Torres-Ortega, P.V. (Pablo Vicente); Smerdou, C. (Cristian); Ansorena-Artieda, E. (Eduardo); et al. "Optimization of a GDNF production method based on Semliki Forest virus vector". European Journal of Pharmaceutical Sciences. (159), 2021, 105726 | es_ES |
dc.identifier.issn | 0928-0987 | - |
dc.identifier.uri | https://hdl.handle.net/10171/63698 | - |
dc.description.abstract | Human glial cell line-derived neurotrophic factor (hGDNF) is the most potent dopaminergic factor described so far, and it is therefore considered a promising drug for Parkinson’s disease (PD) treatment. However, the production of therapeutic proteins with a high degree of purity and a specific glycosylation pattern is a major challenge that hinders its commercialization. Although a variety of systems can be used for protein production, only a small number of them are suitable to produce clinical-grade proteins. Specifically, the baby hamster kidney cell line (BHK-21) has shown to be an effective system for the expression of high levels of hGDNF, with appropriate post-translational modifications and protein folding. This system, which is based on the electroporation of BHK-21 cells using a Semliki Forest virus (SFV) as expression vector, induces a strong shut-off of host cell protein synthesis that simplify the purification process. However, SFV vector exhibits a temperature dependent cytopathic effect on host cells, which could limit hGDNF expression. The aim of this study was to improve the expression and purification of hGDNF using a biphasic temperature cultivation protocol that would decrease the cytopathic effect induced by SFV. Here we show that an increase in the temperature from 33◦C to 37◦C during the “shut-off period”, produced a significant improvement in cell survival and hGDNF expression. Inconsonance, this protocol led to the production of almost 3-fold more hGDNF when compared to the previously described methods. Therefore, a “recovery period” at 37◦C before cells are exposed at 33◦C is crucial to maintain cell viability and increase hGDNF expression. The protocol described constitutes an efficient and highly scalable method to produce highly pure hGDNF. | es_ES |
dc.description.sponsorship | P. Torres thanks the Spanish Ministry of Education (Programa FPU (FPU17/01212)) and Government of Navarra (2019_66_NAB9). E. Garbayo is supported by a “Ramon y Cajal Fellowship (RYC2018-025897-I). MCBB received a Fundación Echébano fellowship. This work was partially supported by the following grants: Instituto Salud Carlos III financed with Feder Funds PI17/01859, Gobierno de Navarra. Departamento de Salud 64/2019 (co-financed at 50% by the European Regional Development Fund through the FEDER Operational Program 2014-2020 of Navarra: “European Union”. | es_ES |
dc.language.iso | eng | es_ES |
dc.publisher | Elsevier | es_ES |
dc.rights | info:eu-repo/semantics/openAccess | es_ES |
dc.subject | GDNF | es_ES |
dc.subject | Biphasic growth, BHK cells, Semliki Forest | es_ES |
dc.subject | Virus | es_ES |
dc.subject | Shut-off | es_ES |
dc.subject | Parkinson's disease | es_ES |
dc.title | Optimization of a GDNF production method based on Semliki Forest virus vector | es_ES |
dc.type | info:eu-repo/semantics/article | es_ES |
dc.description.note | This is an open access article under the CC BY-NC-ND license | es_ES |
dc.identifier.doi | 10.1016/j.ejps.2021.105726 | - |
dadun.citation.number | 159 | es_ES |
dadun.citation.publicationName | European Journal of Pharmaceutical Sciences | es_ES |
dadun.citation.startingPage | 105726 | es_ES |
Files in This Item:
Statistics and impact
Items in Dadun are protected by copyright, with all rights reserved, unless otherwise indicated.