Mouse monoclonal antibodies (MAbs) were raised to dengue-2 virus (D-2V) Cuban (A15) strain and the cell culture supernatants were screened by ELISA using the homologous virus. Three MAb-secreting lines were further characterized using immunoblot, haemaglutination, complement-fixation, and neutralization assays. One of these, MAb 8H8, weakly reacted with the viral nonstructural-1 protein (NS1) but more specifically identified the capsid protein (C), MAb 3E1 recognized in serological assays D-2V A15 but it had a weak reaction to C protein by immunodetection whereas another, MAb 4G3, weakly neutralized the homologous virus isolate and blocked the binding of specific anti-envelope (E) Mab, and its reaction with this protein could not be confirmed using immunoblot assays. These reagents are now being used to compare virulent plus avirulent Caribbean viruses antibody dependent enhancement (ADE) assays.

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An ELISA has been set up for quantifying mouse monoclonal antibodies in culture supernatant. The assay includes rabbit anti-mouse IgG antibodies chromatographycally purified. This preparation was used as coating and as conjugated antibodies in the ELISA. The assay can detect IgG1 with sensitivity of 0.2 ng/mL, IgG2a (0.85 ng/mL), IgG2b (0.13 ng/mL), and IgG3 (3.19 ng/mL) in culture supernatants. The effective working range was from subnanogram per mL quantities to 30 ng/mL by using a computer statistical program. Variation coefficient of ELISA was below 7%. Correlation estimates with a similar ELISA using commercial reagents were performed for each mouse antibody subclass. The assay was able to detect the four mouse monoclonal antibody subclasses in pure human serum as compared with the same ELISA using commercial antibodies. A 24-h pharmacokinetic profile of 1 patient treated with an IgG2a monoclonal antibody is presented.