| Autor: |
Shoji Ideno, Hideki Maeno, Kadue Takahashi, Takeru Urayama, Takamasa Inoue, Kaoru Takeuchi, Kaoru Sakai |
| Rok vydání: |
2021 |
| Předmět: |
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| Zdroj: |
Journal of Virological Methods. 296:114244 |
| ISSN: |
0166-0934 |
| DOI: |
10.1016/j.jviromet.2021.114244 |
| Popis: |
Safety evaluation for the hepatitis E virus (HEV) is required for plasma fractionation products. Plasma-derived HEV (pHEV) is quite unique in that it is associated with a lipid membrane, which, when stripped during manufacturing processes, induces morphological changes in the virus, making it difficult to select proper HEV phenotypes for clearance studies. We developed a convenient system for the preparation of a high titer cell culture-derived HEV (cHEV). In this system, PLC/PRF/5 cells transfected with the wild-type HEV genome generated lipid membrane-associated cHEV for a long period even after cryopreservation. We also examined how this lipid membrane-associated cHEV can be used to verify the robustness of pHEV removal via 19-nm nanofiltration. Sodium-deoxycholate and trypsin (NaDOC/T) treatment not only dissolved lipid but also digested membrane-associated proteins from pHEV and cHEV, making the resulting cHEV particle smaller in size than any pHEV phenotypes generated by ethanol or solvent-detergent treatment in this study. In both 19-nm and 35-nm nanofiltration, cHEV behaved identically to pHEV. These results indicate that cHEV is a useful resource for viral clearance studies in term of availability, and the use of NaDOC/T-treated cHEV ensured robust pHEV removal capacity via 19-nm nanofiltration. |
| Databáze: |
OpenAIRE |
| Externí odkaz: |
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