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. 2021 Oct 4;12:736360. doi: 10.3389/fimmu.2021.736360

Figure 6.

Figure 6

Neutralizing ELISA for neutralizing ability of the antibodies to the three DNA vaccines. The hearts of 20 zebrafish were surgically obtained on the 20th day post the injection of AT730_21605, AT730_22220, AT730_22910, pCDNA3.1(+), and saline solution. Blood was collected and pooled in 3% heparin solution (plasma). Bacterial proteins were separated by SDS-PAGE and transferred to NC membranes. The NC membranes were cut and incubated with the plasma from the animals injected with solution (1), pCDNA3.1(+) (2), and AT730_21605, AT730_22220 or AT730_22910 (3). The membranes were incubated with mouse antiserum against AT730_21605, AT730_22220 or AT730_22910, respectively and then reacted with rabbit antiserum against mouse IgG. Neutralizing rate is calculated by 1 – [(3)/(1)].