Transfer of Anti-Rotavirus Antibodies during Pregnancy and in Milk Following Maternal Vaccination with a Herpes Simplex Virus Type-1 Amplicon Vector

Meier, Anita F.; Suter, Mark; Schraner, Elisabeth M.; Humbel, Bruno M.; Tobler, Kurt; Ackermann, Mathias; Laimbacher, Andrea S.

Serum and milk IgG was analyzed by an indirect ELISA as described previously [50] . Briefly, ELISA plates were coated with sucrose concentrated wt RV (strain Wa, diluted 1:100 in carbonate buffer), washed with PBS-Tween (phosphate buffered saline with 0.05% Tween 20) and incubated with the samples. Serum samples were diluted to 1:5000 and milk samples to 1:500 before application. After washing with PBS-Tween, wells were incubated with the HRP-conjugated goat anti-mouse IgG antibody (Pierce Biotechnologies, Rockford, IL, diluted 1:8000) and analyzed using peroxidase substrate (TMB substrate solution, Thermo Scientific, Waltham, MA). Reaction was stopped with 2 M H 2 SO 4 and the optical density was measured with an ELISA microplate reader. In order to compare values from different ELISA plates, a serum pool of a RV hyper immunized mice from previous studies [17] was used to normalize the values taking the hyper immune sera as 100%.