Author: Meier, Anita F.; Suter, Mark; Schraner, Elisabeth M.; Humbel, Bruno M.; Tobler, Kurt; Ackermann, Mathias; Laimbacher, Andrea S.
Title: Transfer of Anti-Rotavirus Antibodies during Pregnancy and in Milk Following Maternal Vaccination with a Herpes Simplex Virus Type-1 Amplicon Vector Document date: 2017_2_16
ID: 09hmet7r_52
Snippet: HepG2 or Vero2-2 cells were transduced (MOI 2) with the indicated HSV-1 amplicon vector and RVLPs were harvested 48 hpt as described previously [48, 49] . Briefly, cells were scraped into the medium and cell membranes were disrupted by repeated cycles of thawing/freezing. The cell debris was removed by centrifugation at 1400× g and filtration through a 0.45 µm filter. The cleared supernatant was loaded onto a 10% sucrose cushion and concentrate.....
Document: HepG2 or Vero2-2 cells were transduced (MOI 2) with the indicated HSV-1 amplicon vector and RVLPs were harvested 48 hpt as described previously [48, 49] . Briefly, cells were scraped into the medium and cell membranes were disrupted by repeated cycles of thawing/freezing. The cell debris was removed by centrifugation at 1400× g and filtration through a 0.45 µm filter. The cleared supernatant was loaded onto a 10% sucrose cushion and concentrated at 100,000× g for 2 h at 16 • C. For protection, protease inhibitor (protease inhibitor cocktail tablets complete, mini, ethylenediaminetetraacetic acid (EDTA)-free, 1 tablet per 10 mL, Roche Diagnostics, Mannheim, Germany) was added to the supernatant.
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