Fig. 6

Neutralization of ZIKV by mouse serum and monoclonal antibody-containing hybridoma supernatants. a Individual serum samples from five groups of immunized mice (3 mice per group) were assessed for their ability to neutralize ZIKV (strain SD001) using a U937 DC-SIGN flow cytometry-based neutralization assay. The sera were serially diluted five-fold, starting at a 50x dilution. Each point represents the mean of percent neutralization, and the error bars depict the standard deviations. Asterisks indicate a statistical significance achieved at the lowest dilutions of TMV-E and rE samples as compared to adjuvant only sample (P < 0.0001, F test). b) Sub-neutralizing hybridoma supernatants. The ability of fourteen TMV-GL hybridoma supernatants, and two TMV-E supernatants were assessed using a flow-cytometry based neutralization assay. c) and d) . Hybridoma supernatants from c) an rE immunized mouse and d) a TMV-E immunized mouse were serially diluted five-fold starting at a 10x dilution. e) and f) Dot blot assays to detect hybridoma supernatants that bind to TMG-GL. TMV-GL was dotted onto nitrocellulose and probed with hybridoma supernatants from e) an rE immunized mouse or f) a TMV-E immunized mouse, and then detected using horseradish-peroxidase conjugated secondary antibodies and Clarity Western Substrate. g) and h) representative dot blots showing binding patterns of hybridoma supernatants. Hybridoma supernatant #12, from a TMV-E immunized mouse (g), and hybridoma supernatant #26, from an rE immunized mouse (h), were tested for their ability to bind to blotted TMV, TMV-GL, plant-made ZIKV rE (rE), E. coli-made ZIKV rE (rE(−)), TBEV rE, and DENV-2 rE. (−) indicates the lack of glycosylation