Table 4.

Monoclonal antibodies characterization and testing of passive protection

MAbs	Intracellulara	Surface associatedb	IFAc		Affinity KD (nM)d	MAb binding domaine	% Mice survival ratef
			JEV Dengue 2 V
3E10	+	+	+	-	1.1	unknown	16
4 C4	+	+	+	+	56	NS11–143	33
7 C2	+	+	+	+	17	NS1224–352	8
7 H5	+	+	+	+	6.4	NS1224–352	0
8 F1	+	+	+	+	5	NS1224–352	0

^a/bMAbs were incubated with S2-NS1_1–352 cells permeabilized or untreated (intracellular/surface associate) for fluorescence testing by flow cytometry (FACScan).

^c Immunofluorescence assay (IFA) for testing monoclonal antibodies reactivity against BHK cells infected with JEV, and C6/36 cells infected with Dengue 2 virus.

^d The affinity of MAbs to NS1 tested by surface plasmon resonance (SPR).

^e Reactivity of MAbs for S2- NS1_1–143 or NS1_224–352 fragments. Cells lysates were tested by Western blotting (WB). “Unknown” means did not react with both of the NS1 fragments or reduced and boiled NS1.

^f Six to twelve mice were administered with 100μg of 3E10, 4 C4, 7 C2 and 8 F1, respectively, and challenged by JEV SA14.