2.6 Neutralization assays
A pseudotyped EBOV-based neutralization assay was developed to evaluate
the neutralizing activity of different samples. Briefly, 50 μL
pseudotyped EBOV -containing supernatants were incubated with 50 μL
five-fold dilution series of samples at 37 ℃ for 1 h. The pseudotyped
EBOV-sample mixtures were then added onto Huh-7 cells seeded in 96-well
culture plates at an 80-90% monolayer density. Four hours later, the
Huh-7 cells were washed with phosphate-buffered saline (PBS) and
incubated for another 48 h. The Huh-7 cells were lysed using 30 μL/well
of cell lysis buffer (Promega, WI, USA), and 20 μL lysates were used to
determine luciferase activity by adding luciferase substrate (Promega)
into the 96-well luminometer plates. Live EBOV was also applied to
neutralization assay for comparison with the pseudotyped EBOV-based
neutralization assay, which was described previously
(Qiu et al., 2014).