Characterization of endotoxin free protein production of brain-derived neurotrophic factor (BDNF) for the study of Parkinson model in SH-SY5Y differentiated cells
Issued Date
2023-03-01
Resource Type
ISSN
10465928
eISSN
10960279
Scopus ID
2-s2.0-85143849175
Pubmed ID
36481372
Journal Title
Protein Expression and Purification
Volume
203
Rights Holder(s)
SCOPUS
Bibliographic Citation
Protein Expression and Purification Vol.203 (2023)
Suggested Citation
Saisawang C., Priewkhiew S., Wongsantichon J., Reamtong O., Nopparat C., Mukda S., Ketterman A.J., Govitrapong P. Characterization of endotoxin free protein production of brain-derived neurotrophic factor (BDNF) for the study of Parkinson model in SH-SY5Y differentiated cells. Protein Expression and Purification Vol.203 (2023). doi:10.1016/j.pep.2022.106212 Retrieved from: https://repository.li.mahidol.ac.th/handle/20.500.14594/81647
Title
Characterization of endotoxin free protein production of brain-derived neurotrophic factor (BDNF) for the study of Parkinson model in SH-SY5Y differentiated cells
Other Contributor(s)
Abstract
Human neuronal cells are a more appropriate cell model for neurological disease studies such as Alzheimer and Parkinson's disease. SH-SY5Y neuroblastoma cells have been widely used for differentiation into a mature neuronal cell phenotype. The cellular differentiation process begins with retinoic acid incubation, followed by incubation with brain-derived neurotrophic factor (BDNF), a recombinant protein produced in E. coli cells. Endotoxin or lipopolysaccharide (LPS) is the major component of the outer membrane of bacterial cells that triggers the activation of pro-inflammatory cytokines and ultimately cell death. Consequently, any endotoxin contamination of the recombinant BDNF used for cell culture experiments would impact on data interpretation. Therefore, in this study, we expressed the BDNF recombinant protein in bacterial endotoxin-free cells that were engineered to modify the oligosaccharide chain of LPS rendering the LPS unable to trigger the immune response of human cells. The expression of DCX and MAP-2 in differentiated cells indicate that in-house and commercial BDNF are equally effective in inducing differentiation. This suggests that our in-house BDNF protein can be used to differentiate SH-SY5Y neuroblastoma cells without the need for an endotoxin removal step.