Table 5. Epitope Specific Proportions of Envelope Protein Specific IgM and IgG from DENV-2 Infected Human Sera.

	DENV-2 Antigen1	Epitope Specific Target2	Mean Endpoint Titer3	Endpoint Titer Range	Range of Percent Response4	Mean (Median) Percent Epitope Specific Response
IgM
	WT	All	2.99×105	2.14×104–1.51×106	100	100
	G106R-L107D*†	EDIIFP	1.66×105	6.07×102–4.79×105	<1–97	44 (34)
	K310E-E311R-P364R†	EDIIICR	1.71×105	1.59×104–8.32×105	<1–90	43 (39)
	G106R-L107D-K310E-E311R-P364R*	EDIIFP−EDIIICR	1.26×105	5.00×102–3.98×105	<1–91	58 (67)
	K305E-K388D	EDIIITS	1.79×104	2.19×102–7.98×104	<1–20	6 (4.5)
IgG
	WT*†	All	2.40×106	1.24×104–1.10×107	100	100
	G106R-L107D*	EDIIFP	1.54×106	7.37×102–4.79×106	<1–94	36 (36)
	K310E-E311R-P364R†	EDIIICR	2.46×106	1.16×104–1.05×107	<1–26	5.8 (13)
	G106R-L107D-K310E-E311R-P364R	EDIIFP−EDIIICR	6.76×105	4.74×102–1.70×106	<1–96	72 (64)
	K305E-K388D	EDIIITS	2.40×104	1.24×102–4.50×105	<1–8	1 (<1)

¹The mutant antigens used are all knock-out mutants. The symbols * and † depict significantly different mean endpoint titers determined with antigens sharing the same symbol. Mean endpoint titers were considered significantly different when the 95% confidence interval for the difference between the means did not cross zero.

²Epitope specific antibody populations targeted by the different knock-out antigens. WT antigen measures antibody recognizing all E-protein epitopes. EDII_FP denotes broadly cross-reactive epitopes incorporating the E-protein structural domain II fusion peptide. EDIII_CR denotes predominately complex cross-reactive epitopes incorporating residues within E-protein structural domain III. EDII_FP−EDIII_CR denotes individual or overlapping epitopes incorporating either or both the EDII fusion peptide or EDIII. EDIII_TS denotes EDIII DENV-2 type-specific epitopes and were determined by the reactivity difference between the K305E and K388D antigens (see methods for details).

³Endpoint titers determined with the knock-out antigen, thus representing immunoglobulins recognizing epitopes not targeted by the knock-out antigen. Because the EDIII DENV-2 type-specific response was calculated as the percent difference between K305E and K388D reactivities, the titers for EDIII_TS were calculated as the WT titers multiplied by the percent EDIII_TS response.

⁴Because the mutant antigens knock-out antibody recognition of specific epitopes, the percent of immunoglobulin recognizing a particular epitope was determined for each individual sera by calculating the percent reactivity measured with a mutant antigen relative to that determined with the WT antigen and subtracting this value from 1.0; (1- [Endpoint_mutant/Endpoint_wt])×100; for the percent DENV-2 specific reactivity we used (1-[(Endpoint_K305E/Endpoint_WT)−(Endpoint_K388D)/Endpoint_WT)])×100.