Table 1. Characterization of mAb activities against DENV4 by ELISA, IFA, WB, and PRNT50.
mAbs | Isotyping | Class a | ELISA b | IFA c | WB d | Specificity e | DENV4 PRNT50 (μg/ml) f | |||||||||
---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|
D1 | D2 | D3 | D4 | D1 | D2 | D3 | D4 | D1 | D2 | D3 | D4 | |||||
DD1-4 | IgG2a, κ | Type | - | - | - | + | - | - | - | + | - | - | - | + | EDI-II | >40 |
DD3-7 | IgG1, κ | Type | - | - | - | + | - | - | - | + | - | - | - | + | NS1 | n.d. |
DD4-3 | IgG2a, κ | Type | - | - | - | + | - | - | - | + | - | - | - | + | E | >40 |
DD5-1 | IgG1, κ | Type | - | - | - | + | - | - | - | + | - | - | - | + | E | >40 |
DD7-8 | IgG2a, κ | Subcomplex | + | - | - | + | + | - | - | + | + | - | - | + | NS1 | n.d. |
DD9-4 | IgG2a, κ | Type | - | - | - | + | - | - | - | + | - | - | - | + | E | >40 |
DD11-4 | IgG1, κ | Complex | + | + | + | + | + | + | + | + | + | + | + | + | EDI-II | >40 |
DD13-4 | IgG1, κ | Type | - | - | - | + | - | - | - | + | - | - | - | + | EDI-II | >40 |
DD14-1 | IgG2a, κ | Subcomplex | - | - | + | + | - | - | + | + | - | - | + | + | EDI-II | >40 |
DD15-2 | IgG2b, κ | Subcomplex | - | - | + | + | - | - | + | + | - | - | + | + | E | >40 |
DD17-4 | IgG2a, κ | Type | - | - | - | + | - | - | - | + | - | - | - | + | EDIII | >40 |
DD18-5 | IgG2a, κ | Complex | + | + | + | + | + | + | + | + | + | + | + | + | EDI-II | ≧20 |
DD27-8 | IgG2a, κ | Type | - | - | - | + | - | - | - | + | - | - | - | + | EDIII | >40 |
DD30-4 | IgG2a, κ | Type | - | - | - | + | - | - | - | + | - | - | - | + | EDI-II | >40 |
DD31-3 | IgG1, κ | Type | - | - | - | + | - | - | - | + | - | - | - | + | EDI-II | >40 |
DD33-2 | IgG1, κ | Subcomplex | - | + | - | + | - | + | - | + | - | + | - | + | NS1 | n.d. |
a The classes of mAbs include complex-reactive, subcomplex-reactive, and type-specific. The results were determined by ELISA, IFA, and WB.
b Plates coated with rabbit hyper-immune sera against DENV1-4 were used to test the reactivity of mAbs. (+) The OD values above cut off value were positive (+), and those below were negative (-).
c The IFA results using DENV1-4-infected BHK-21 cells were confirmed as described in the Methods.
d Western blotting (WB) using cell lysates from DENV1-4-infected C6/36 cells was performed as described in the Methods.
e The specificities of mAbs were determined using lysates of virus-infected cells. The binding domains of mAbs were identified by recombinant proteins. E: envelope protein; EDI-II, recombinant domain I-II of E protein; EDIII, recombinant domain III of E protein; NS1, nonstructural protein 1.
f The PRNT50 values are shown as the lowest concentration of mAbs that reduced ≧50% of plaques. (n.d.) not determined.