Epitope reactions can be gauged by relative antibody discriminating specificity (RADS) values supported by deletion, substitution and cysteine bridge formation analyses: potential uses in pathogenesis studies

Table 2 Mouse PAb reactions against DENV-2 NS1 glycoprotein peptides prepared in the natural and reverse orientations

Peptide/Epitope^a	N/R^b	Sequence^c	Average ELISA absorbance (492 nm) of PAbs^e
			C57BL or B10 mouse haplotype anti-DENV-2 NS1glycoprotein PAbs^d
C57				S	RIII	G	BR	D2N	A	S(C)
N-TERM	+	5-VVSWKNKELK C-15	0.87	0.92	0.78	0.71	0.67	0.70	0.68	0.18
N-TERM	-	15-CKLEK NKWSVV-5	0.56	0.58	0.54	0.56	0.43	0.51	0.55	0.14
LD2	+	25-VHTWTEQYK-33	0.67	0.54	1.21	1.32	0.69	0.72	1.02	0.27
LD2	-	33-KYQETWTHV-25	0.44	0.37	0.83	1.24	0.26	0.45	0.77	0.25
24A	+	61-TRLE NLMWK-69	0.80	0.77	0.79	0.56	0.43	0.45	0.46	0.28
24A	-	69-KWMLNELR T-61	0.35	0.32	0.52	0.42	0.41	0.28	0.40	0.25
N'LX1	+	109-TELR YSWKT-117	1.23	1.41	1.01	0.97	1.14	1.16	0.96	0.27
N'LX1	-	117-TKWSYRLE T-109	0.98	0.86	0.82	0.88	0.99	1.06	0.89	0.22
LX1	+	113-YSWKTWGKA-121	0.87	1.08	0.44	0.23	0.30	1.16	0.21	0.13
LX1	-	121-AKGWTKWSY-113	0.28	0.39	0.63	0.42	0.34	0.34	0.36	0.18
LX2/1	+	209-TWKIEK ASF-217	0.72	0.68	1.04	1.01	0.72	0.50	0.68	0.20
LX2/1	-	217-FSAKEIK WT-209	0.81	1.20	1.15	1.03	1.14	0.65	0.61	0.24
LX2/2	+	267-PWHLGKLE M-275	0.92	0.79	1.02	1.04	0.76	0.68	0.63	0.21
LX2/2	-	275-MELK GLHWP-267	0.98	1.03	1.06	1.12	0.82	0.54	0.67	0.18
24C	+	301-TTASGKLIT-309	0.65	0.74	0.61	0.66	0.47	0.40	0.43	0.13
24C	-	309-TILKGSATT-301	0.41	0.47	0.51	0.56	0.42	0.39	0.48	0.12
LX2/3	+	331-YGMEIR PLK-339	0.65	0.71	1.24	1.01	0.69	0.81	0.62	0.26
LX2/3	-	339-KLPRIE MGY-331	0.61	1.11	1.21	1.16	0.51	0.66	0.59	0.20

^aName of the epitope or MAb 1G5.4-A1-C3 and/or 1C6.3 defined N’LX1, LX2/1, LX2/2 and LX2/3 peptide sequences that contained ELK/KLE-type or KELK/KLEK-type motifs.
^bSynthetic peptide sequences prepared in either the natural (N: +) or reverse (R: -) orientations.
^cSequence position and amino acid sequence of the DENV-2 NS1 glycoprotein represented by synthetic peptides prepared both the natural and reverse orientations. The ELK/KLE-type and KELK/KLEK-type motifs are underlined.
^dGroups of C57BL (C57) (H-2^b) or congeneic (H-2) B10 mouse strains (B10.S (S), B10.RIII (RIII), B10.BR (BR), B10.D2N (D2N) and B10.A (A)) immunized with the native hexameric DENV-2 e/sNS1 glycoprotein (gp), or the control protein (ovalbumin) (B10.S(C): (S(C)).
^eAverage ELISA absorbance values of mouse PAb pools reacted with duplicate sets of synthetic peptides (standard deviation range: 0.00 (low values) to 0.04 (high values)). PAb absorbance (Abs) values which were ≥ 0.650 against the peptide prepared in the natural orientation that showed ≥ 1.50-fold or ≥ 2.50-fold RADS values (Abs against the natural (+)/reverse orientated peptide sequence), are underlined and shown in bold, respectively.