Cat. No.
MABL-1466
Application
crystallization, neutralizing, SPR, ELISA
Isotype
Engineer antibody
Species Reactivity
Dengue Virus
Clone No.
2C8
From
Recombinant Antibody
Specificity
The antibody binds to domain III of the envelope (E) protein of Dengue virus. In particular, it recognizes the N-terminal region of EDIII (DIII-DI hinge residues 298-303) and the BC loop (residues 328-334), and residues K361, V282, E383 and P384. 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.
Alternative Names
E-protein; DENV EDIII; Dengue envelope protein domain III; E protein domain III; EDIII; DVC-21; DV 21; DV-21; Envelope protein E
UniProt
W5RZ25
Immunogen
The original antibody was generated by immunizing mice with DENV2 16681.
Application Notes
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 antibody (IgG2a) with 50% neutralization titers in the sub-nanomolar range (50% FRNT at 0.025 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 has strong capacity to promote antibody dependent enhancement (ADE). ADE was analysed using U937 cells, the antibody demonstrated typical infection enhancement with peak titers of over 1000-fold enhancement over background and a wide range of concentrations at which enhancement occurred. The humanized version of the antibody (human IgG1) showed 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= 176.7 nM) (Renner et al., 2018; PMID: 30323338).
Antibody First Published
Renner et al. Characterization of a potent and highly unusual minimally enhancing antibody directed against dengue virus. Nat Immunol. 2018 Nov;19(11):1248-1256. PMID:30323338
Note on publication
The paper describes the generation and characterization of the antibody, which is specific for the envelope protein of Dengue virus. The antibody shows neutralizing activity, but promotes antibody dependent enhancement.
Size
100 μg Purified antibody.
Concentration
1 mg/ml.
Purification
Protein A affinity purified
Buffer
PBS with 0.02% Proclin 300.
Storage Recommendation
Store at 4⁰C for up to 3 months. For longer storage, aliquot and store at - 20⁰C.
