TABLE 3.
Cross-reactions of MAbs generated against the DENV-2 NS1 glycoprotein with synthetic peptides representing surface-exposed regions of DENV-2 E glycoproteins
| Peptide sequencea | DENV-2 genotype(s) (strains)b | Average MAb ELISA absorbance (% of maximum absorbance)c
|
|||||
|---|---|---|---|---|---|---|---|
| 1H7.4 | 5H4.3 | 3D1.4 | 1G5.3 | 1G5.4.A1.C3 | 1C6.3 | ||
| 44-ELIKTEAKQP-53 | Nearly all (TR, NG-C, M2) | 0.03 | 0.08 | 0.06 | 0.08 | 0.55 | 0.37 |
| 48-TEAKQPATLR-57 | Nearly all (TR, NG-C, M2) | 0.10 | 0.07 | 0.03 | 0.25 | 0.43 | 0.29 |
| 67-NTTTESRCPT-76 | As1, As2, Cos, Am/As (M2) | 0.04 | 0.04 | 0.05 | 0.15 | 0.92 (80.7) | 0.53 |
| 67-NTTTDSRCPT-76 | Am, some As2 (TR, NG-C) | 0.04 | 0.09 | 0.06 | 0.23 | 0.52 | 0.32 |
| 67-NTTTASRCPT-76 | Cos | 0.03 | 0.07 | 0.04 | 0.45 | 0.23 | 0.22 |
| 74-CPTQGEPSLN-83 | Nearly all (TR, NG-C, M2) | 0.04 | 0.05 | 0.03 | 0.03 | 0.06 | 0.05 |
| 74-CPTLGEPSLN-83 | A few As1, Am/As, Cos | 0.03 | 0.04 | 0.03 | 0.03 | 0.02 | 0.04 |
| 93-KHSMVDRGW-101 | All (TR, NG-C, M2) | 0.06 | 0.16 | 0.10 | 0.22 | 0.11 | 0.09 |
| 95-SMVDRGWGN-103 | All (TR, NG-C, M2) | 0.04 | 0.21 | 0.13 | 0.21 | 0.24 | 0.13 |
| 97-VDRGWGNGC-105 | All (TR, NG-C, M2) | 0.04 | 0.04 | 0.15 | 0.81 (20.8) | 0.48 | 0.25 |
| 99-RGWGNGCGL-107 | All (TR, NG-C, M2) | 0.04 | 0.06 | 0.07 | 0.38 | 0.24 | 0.21 |
| 101-WGNGCGLFG-109 | All (TR, NG-C, M2) | 0.02 | 0.11 | 0.13 | 0.67 (17.3) | 0.21 | 0.14 |
| 103-NGCGLFGKG-111 | All (TR, NG-C, M2) | 0.01 | 0.23 | 0.06 | 0.16 | 0.10 | 0.08 |
| 127-GKIVQPENLE-136 | Many (TR, M2) | 0.03 | 0.06 | 0.10 | 0.57 | 0.21 | 0.15 |
| 156-GKHGKEIKIT-165 | As1, As2, Cos, Am/As (NG-C, M2) | 0.03 | 0.02 | 0.14 | 0.67 (17.3) | 0.94 (82.5) | 1.06 (86.9) |
| 156-GKHGKEVKIT-165 | Am (TR) | 0.04 | 0.02 | 0.10 | 0.48 | 0.64 | 0.98 (80.3) |
| 273-SSGNLLFTGH-282 | All (TR, NG-C, M2) | 0.13 | 0.08 | 0.52 | 1.47 (37.9) | 0.16 | 0.08 |
| 273-SSGNL-277 | All (TR, NG-C, M2) | 0.03 | 0.03 | 0.05 | 0.69 (17.8) | 0.06 | 0.06 |
| 288-RMDKLQLKG-296 | All (TR, NG-C, M2) | 0.10 | 0.04 | 0.06 | 0.15 | 0.61 | 0.23 |
| 304-GKFKVVKEIA-313 | As1, As2, Cos, Am (TR, NG-C) | 0.04 | 0.05 | 0.09 | 0.18 | 0.22 | 0.36 |
| 304-GKFKIVKEIA-313 | Am/As, one Am | 0.05 | 0.04 | 0.10 | 0.14 | 0.23 | 0.32 |
| 304-GKFKVVEEIA-313 | A few As1 (M2) | 0.03 | 0.03 | 0.04 | 0.07 | 0.42 | 0.43 |
| 338-EIMDLEKRHV-347 | Nearly all (TR, NG-C) | 0.03 | 0.03 | 0.05 | 0.05 | 0.34 | 0.26 |
| 338-EIMDLDNRHV-347 | One As1 (M2) | 0.03 | 0.03 | 0.06 | 0.05 | 0.80 (70.2) | 0.28 |
| 385-GQLKLNWFKK-394 | Nearly all (TR, NG-C, M2) | 0.07 | 0.25 | 0.10 | 0.03 | 0.15 | 0.23 |
Amino acid sequences of synthetic peptides from DENV-2 E glycoproteins. Surface-exposed sequences are underlined, and amino acid substitutions in the different DENV-2 strains are shown in bold.
Sequences are present in the E glycoproteins of all, nearly all, many, a few, or one DENV-2 strain of the Asian 1 (As1), Asian 2 (As2), American/Asian (Am/As), American (Am), and Cosmopolitan (Cos) genotypes and present in the mildly pathogenic Trinidad 1751 (TR) (American), moderately pathogenic New Guinea-C (NG-C) (Asian 2), or highly pathogenic Malaysia M725 (M2) (Asian 1) DENV-2 strain (in parentheses).
Average ELISA absorbances (492 nm) of MAbs diluted to 100 times their ELISA titer (1/log10t50) obtained against the DENV-2 NS1 glycoprotein and incubated with each synthetic peptide in duplicate (standard deviation range, 0.00 to 0.02). Values above the absorbance cutoffs of 0.650 and 1.000 are shown in bold and underlined, respectively, and the percentages of maximum absorbance obtained against the 25-33, 61-69, 113-119, 301-309, 109-117, and 208-216 peptide sequences from the DENV NS1 glycoprotein, using MAb 1H7.4 (314 ng/ml), MAb 5H4.3 (1,714 ng/ml), MAb 3D1.4 (442 ng/ml), MAb 1G5.3 (575 ng/ml), MAb 1G5.4-A1-C3 (1,595 ng/ml), and MAb 1C6.3 (4,439 ng/ml), respectively (Table 2), are shown in parentheses.