Phenotypic characterization of cell culture-derived hepatitis E virus subjected to different chemical treatments: Application in virus removal via nanofiltration

• Published in: Journal of virological methods Vol. 296; p. 114244
• Main Authors: Ideno, Shoji, Inoue, Takamasa, Takahashi, Kadue, Urayama, Takeru, Maeno, Hideki, Takeuchi, Kaoru, Sakai, Kaoru
• Format: Journal Article
• Language: English
• Published: Elsevier B.V 01.10.2021
• Subjects: Hepatitis E virus; Nanofiltration; Plasma fractionation; Viral clearance; Viral clearance; TBS; NaDOC/T; Plasma fractionation; PA-GM; S/D; A549-MM; TB; Hepatitis E virus; LRV; Nanofiltration; P/S; PLC-MM; HEV; cHEV; JB; fHEV; pHEV
• ISSN: 0166-0934; 1879-0984
• DOI: 10.1016/j.jviromet.2021.114244

[Display omitted] •Lipid-associated, cell culture-derived HEV (cHEV) was obtained at 9 log copies/mL.•cHEV behaves identically to plasma-derived HEV (pHEV) during nanofiltration.•Lipid-stripped cHEV can be used to verify robust pHEV removal via nanofiltration. 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.