TABLE 5 

Epitope mapping of DENV-specific FL region binding human MAbs by binding assays to WNV E protein FL or bc loop mutants or shotgun mutagenesis studies with DENV E protein

Type of infectionSubject
no.
MAbBinding phenotype in ELISA as + or − to the
indicated West Nile virus E protein
DENV E protein
mutation(s) that
disrupts binding
Wild-type E
proteind
QUAD
mutantd
W101R
mutantd
Mutations that disrupted
binding in DENV2 E
yeast display (W101R)
G106R, or L107D)
Primary DENV1
vaccine
491M12.2NDa101, 106, 107R99A, W101A, L107A, F108A, G111A
Primary DENV1
wild type
1061M6.2+++NDW101A, L107A
2J21ND101, 106, 107G78A, E79A, L107A, K110A, G111A
2I23+ND101, 106, 107Not determinedb
101M4+++NDW101R, L107P, L107R, G111R
Primary DENV2
wild type
195M22ND101, 106, 107K26A (prM, D4); N103D, G104E, G111R (D3)
Primary DENV3
wild type
31I17ND101, 106, 107W101A, G106A
2A15++NDW101A
1181M7+++NDW101R, W101C, G111R
Secondary
wild type
92C7+++NDW101R, W101G, W101C, L107P, L107R, F108I, G111R (D3); W101A, F108A (D4)
10891N5++NDW101R, L107P, L107R, G111R
33871F12.2+NDW101A, F108A
1C19NAcNo disruption detectedR73A, G78A (D4); R73Q, G78D, and E79V (D3)
1153H4++NDW101A
1841E4++NDW101A, F108A
1B19+NDW101A
1C18+NDW101R, W101C, L107P, L107R, G111R
1H20+ND101, 106, 107R99A, W101A, L107A, F108A
1I16+NDW101A, F108A
1K16+101, 106, 107L107P, L107R, G111R
1L4+NDW101A
1L6++NDG100A, W101A, F108A
1N8+NDW101R, W101G, W101C, L107P, L107R, G111R
2M11+NDW101A, G104A
3B4++NDW101A, F108A
3D18++NDW101A, F108A
3G5+NDF108A
4E8+NDG100A, W101A, F108A
5C8+NDW101A, F108A
5K17+NDW101A, G104A, G106A
ControlE16 control+++++++++NDND
E18 FL-specific control+++NDND
  • a ND, not done.

  • b Not determined because MAb 2I23 exhibited a high level of nonspecificity and so it was not mapped.

  • c NA indicates not applicable. Since 1C19 did not bind the wild-type WNV protein, loss of binding to the W101R mutant was not tested.

  • d Optical densities at A450 are summarized as follows: –, ≤0.4; +, 0.5 to 1.2; ++, 1.3 to 2.0; +++, ≥2.