The integrity of chemically treated plasmid DNA as a chemical-based choice for prion clearance

Nozomi Takada, Yasuharu Niwa, Tomoaki Teshigawara, Kazue Isogai, Hanayuki Okura, Akifumi Matsuyama

Research output: Contribution to journalArticlepeer-review

1 Citation (Scopus)

Abstract

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.

Original languageEnglish
Pages (from-to)112-120
Number of pages9
JournalRegenerative Therapy
Volume15
DOIs
Publication statusPublished - 12-2020

All Science Journal Classification (ASJC) codes

  • Biomaterials
  • Biomedical Engineering
  • Developmental Biology

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