The integrity of chemically treated plasmid DNA as a chemical-based choice for prion clearance
| Autor: | Akifumi Matsuyama, Yasuharu Niwa, Hanayuki Okura, Tomoaki Teshigawara, Nozomi Takada, Kazue Isogai |
|---|---|
| Rok vydání: | 2020 |
| Předmět: |
0301 basic medicine
BSE bovine spongiform encephalopathy SDS sodium dodecyl sulfate TSE transmissible spongiform encephalopathy Biomedical Engineering TCA trichloroacetic acid Green fluorescent protein Biomaterials Gdn-SCN guanidine-thiocyanate 03 medical and health sciences chemistry.chemical_compound 0302 clinical medicine Plasmid dna Gdn-HCl guanidine hydrochloride medicine Prion inactivation lcsh:QH573-671 Regenerative medical products lcsh:R5-920 NaOH sodium hydroxide Transmissible spongiform encephalopathy lcsh:Cytology Chemistry Manufacturing process CJD Creutzfeldt-Jakob Disease Transfection medicine.disease nervous system diseases Standard for biological raw materials 030104 developmental biology Biochemistry Reagent Original Article lcsh:Medicine (General) 030217 neurology & neurosurgery Function (biology) DNA Developmental Biology |
| Zdroj: | Regenerative Therapy Regenerative Therapy, Vol 15, Iss, Pp 112-120 (2020) |
| ISSN: | 2352-3204 |
| DOI: | 10.1016/j.reth.2020.05.005 |
| Popis: | In regenerative medical products for clinical applications, a major concern is the risk of ruminant-derived materials developing transmissible spongiform encephalopathy (TSE) in the manufacturing process. Because of the risk of TSE causing prion disease, the raw materials derived from ruminants should be compliant with the “Standard for Biological Raw Materials” to ensure the quality and safety of pharmaceutical products. We therefore tested whether plasmid DNA could withstand four chemical reagents (Gdn-HCl, Gdn-SCN, TCA, or SDS), having referred to the report by Tateishi et al. [1], which describes how Creutzfeldt–Jakob disease pathogens can be inactivated by chemical reagents capable of producing a 7-log reduction in prion inactivation. We observed that plasmid DNA was mixed with chemical reagents and that the functionality of plasmid DNA was equivalent for both chemical and non-chemical treatment. The potency of plasmid DNA was monitored by the existence of DNA fragments and the function by which GFP proteins were produced by HEK293-cell transfected plasmid DNA. The existence of DNA fragments was detected in plasmid DNA treated by chemical reagents, except when undergoing TCA treatment. Additionally, when HEK293 cells were transfected with the plasmid DNA after chemical treatment, GFP protein was produced. These results indicate that plasmid DNA can withstand the chemical treatments for blocking prion transmission. Highlights • Inactivation methods need to be carefully chosen based on the raw materials. • Plasmid DNA withstood chemical treatment, undergoing Gdn-HCl, Gdn-SCN, and SDS for prion inactivation. • The integrity of chemically treated plasmid DNA is not compromised as a result of the treatment. |
| Databáze: | OpenAIRE |
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