Protective immunity to Japanese encephalitis virus associated with anti-NS1 antibodies in a mouse model

Table 4 Monoclonal antibodies characterization and testing of passive protection

MAbs	Intracellular^a	Surface associated^b	IFA^c		Affinity KD (nM)^d	MAb binding domain^e	% Mice survival rate^f
			JEV Dengue 2 V
3E10	+	+	+	-	1.1	unknown	16
4 C4	+	+	+	+	56	NS1_1–143	33
7 C2	+	+	+	+	17	NS1_224–352	8
7 H5	+	+	+	+	6.4	NS1_224–352	0
8 F1	+	+	+	+	5	NS1_224–352	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.

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