Anti-EDIII of DENV2 mAbs [3H5](MABL-1467)

MABL-1467

crystallization, in vivo, neutralizing, SPR, ELISA, FC

Engineer antibody

Dengue Virus

3H5

Recombinant Antibody

The antibody binds to domain III of the envelope (E) protein of Dengue virus. In particular, it targets residues buried between E dimers and located close to the viral membrane. The antibody is serotype specific to DENV2. The E protein is a structural protein located on the viral surface and consists of three distinct domains termed EDI, EDII, and EDIII. Of these, EDIII mediates attachment to target cells, prior to internalization into host endosomes.

E-protein; DENV EDIII; Dengue envelope protein domain III; E protein domain III; EDIII; DVC-21; DV 21; DV-21; Envelope protein E

W5RZ25

The original antibody was generated by immunizing mice with DENV2 strain New Guinea C (NGC).

Binding avidity/affinity of both Full length (IgG) and Fab fragments of the antibody to whole virion measured by ELISA. The crystal structure of the Fab fragment in complex with EDIII of DENV2 NGC was determined. Focus reduction neutralization assays showed highly potent neutralization of DENV2 NGC and 16681 for the IgG1 fragment with 50% neutralization titers in the sub-nanomolar range (50% FRNT at 0.044 nM ). Pre- and postattachment neutralization assays showed the antibody is capable of significantly inhibiting infection after the virus had bound to the cells. The antibody promotes little antibody dependent enhancement (ADE). ADE was analyzed using U937 cells, the antibody showed no enhancing capacity for DENV2 NGC and reduced enhancement of DENV2 16681 at a very narrow concentration range. Also the IgG2a format of the antibody showed no enhancement of infection for DENV2 NGC while still potently neutralizing infection. The humanized version of the antibody (human IgG1) showed little enhancement for DENV2/16681 and DENV2/NGC. Surface plasmon resonance assay was performed to evaluate the binding specificity and affinity between the antibody and recombinant EDIII (Kd= 1,0 nM). The antibody showed potential as therapeutic (Renner et al., 2018; PMID: 30323338). Antibody-dependent infection enhancement (ADE) was reported for the antibody using P-388D1 mouse macrophage-like cells (Morens et al., 1987; PMID:3504546). This antibody was used for flow cytometry on wild-type DIII and mutant DIII expressed on the yeast surface (Sukupolvi-Petty et al., 2007; PMID: 17881453). The antibody was reported to cause ADE in THP-1 cells at low doses however, it did not enhance DENV-2 infection of AG129 mice. Fifty-percent plaque reduction neutralization test (PRNT50) values of the antibody determined by using 100 PFU of S221 (PRNT50 values of 0.19 ug) (Watanabe et al., 2015; PMID: 25787279). The antibody was labeled with Eu* conjugate and used in a sandwich ELISA for the detection of DENV-2 (Shukla et al., 2018; PMID: 29367852)

Gentry et al. Identification of distinct antigenic determinants on dengue-2 virus using monoclonal antibodies. Am. J. Trop. Med. Hyg. 31:548– 555. PMID:6177259

The paper describes the generation and characterization of the antibody, which is specific for Dengue virus.

100 μg Purified antibody.

1 mg/ml.

Protein A affinity purified

PBS with 0.02% Proclin 300.

Store at 4⁰C for up to 3 months. For longer storage, aliquot and store at - 20⁰C.