Figure 6

Detection sensitivities of MAb 2C4.6 against the DENV s/e NS1 glycoproteins in a 3 h assay. The maximum detection sensitivity of the assay was assessed against the same serial dilutions of immunoaffinity-purified s/e NS1 glycoproteins when the total dot-blot assay time was reduced to 3 h. For this assay, serial two-fold dilutions from 512 to 2 ng/ml of immunoaffinity-purified s/e NS1 glycoprotein of each DENV serotype (DENV-1 to DENV-4) or buffer only (0), prepared in undiluted human sera were added at 10 μl volumes to paired 5-mm squares marked on a nitrocellulose membrane. After being dried and treated with 3% H₂O₂/H₂O for 5 min to quench the endogenous human serum peroxidases, this membrane was washed and blocked before being processed by sequential reaction steps with MAb 2C4.6, peroxidase-labelled goat anti-mouse IgG (H&L) and 4-chloro-1-naphthol/3,3’diaminobenzidine (CND) substrate. The results were obtained from one of duplicate experiments performed that gave similar results.