High Environmental Stability of Hepatitis B Virus and Inactivation Requirements for Chemical Biocides

Than, Thoa Thi; Jo, Eunji; Todt, Daniel; Nguyen, Phuong Hong; Steinmann, Jochen; Steinmann, Eike; Windisch, Marc P

HBV particles were produced in the HepAD38 cell line, harboring an over-length HBV genome under the control of a tetracycline-repressed promoter element (the tet-off system) [7] . The production process is described elsewhere [8] . Briefly, HepAD38 cells were cultured in Dulbecco's modified Eagle's medium/F-12 supplemented with 10% fetal bovine serum (FBS), 50 U/mL penicillin 50 µg/mL streptomycin, 400 µg/mL Geneticin, 5 µg/mL insulin, 50 µM hydrocortisone hemisuccinate and 0.3 µg/mL tetracycline for 2 weeks. After reaching cell confluency, tetracycline was removed from the medium to induce virus production. Cell culture supernatants were collected once weekly, filtered with a Millipore sterile vacuum filter (0.2 µm), and stored at 4°C.