An ELISA plate was coated with 2ug/ml of NS1 antigen per well, then blocked with 5% BSA. Primary antibody was titrated as shown in the figure below, starting from a concentration of 1ug/ml, and the detection antibody used was Goat anti-human IgM:HRP (Bio-Rad, 1:8000). The substrate used was TMB (KPL).
An ELISA plate was coated with 10ng and 100ng of NS1 antigen per well, then blocked with 1% BSA. Primary antibody was used at a concentration of 1ug/ml, and the detection antibody used was Goat anti-mouse IgG:HRP (Bio-Rad, 1:2000). The substrate used was TMB (KPL). The assay confirmed high specificity for Zika virus NS1; there was no cross-reactivity with NS1 from Dengue virus serotypes 1-4 (DENV), West Nile virus (WNV), Yellow Fever virus (YFV), Japanese Encephalitis virus (JEV) and Tick-borne Encephalitis virus (TBE).
HUMAN ANTI ZIKA VIRUS NS1 IGM MONOCLONAL ANTIBODY (B4)
Human IgM anti Zika virus NS1 monoclonal antibody is specific for the NS1 protein of Zika virus, detecting NS1 from both the Uganda and Suriname strains. It demonstrates negligible cross-reactivity with NS1 proteins from Dengue virus (all serotypes), Japanese Encephalitis Virus and Yellow Fever Virus. A small amount of cross-reactivity has been observed with the NS1 protein from West Nile Virus in a direct ELISA.
This human IgM anti Zika virus NS1 monoclonal antibody has been prepared by chimerization from the mouse monoclonal antibody (clone B4). The original variable domains of this antibody have been retained, whilst the constant regions have been replaced with human IgM.
The antibody is designed to provide a control for assays in which human serum is tested for antibodies specific for Zika NS1 protein, which will result from the immune response following an infection with Zika virus. The antibody has been tested in ELISA using recombinant Zika NS1 protein as the target antigen.