The utilization of biotin-antibiotin interaction for the detection of antibody to hepatitis B surface antigen (anti-HBs).

A biotin-antibiotin solid-phase enzyme-linked immunoassay for the detection and quantitation of the antibody to hepatitis B surface antigen (anti-HBs) is described. The assay utilizes hepatitis B surface antigen as a solid-phase 'capture' reagent and a mixture of biotinylated HBsAg and antibiotin-conjugated horseradish peroxidase as a detector reagent. The assay was compared to a commercial enzyme immunoassay (AUSAB EIA) which used the biotin-avidin system for anti-HBs detection. The two assays were found to measure the same molecules and to correlate well regarding anti-HBs titers.

Preparation and characterization of monoclonal antibodies to rotavirus.

By utilizing a strain of cultivable simian rotavirus (SA-11) as an immunizing antigen, we prepared 4 clones of mouse-mouse hybridoma, namely C127, C139, C172, and C214 which secreted monoclonal antibodies against the immunogen itself, SA-11 and also against other group A strains such as Wa and S2. Western blot analyses revealed that all of these antibodies are directed against VP6, a 42 kDa major inner capsid protein of group A rotavirus. Competitive experiments suggested that C127, C172 and C214 recognized three distinct epitopes on VP6, while C139 appeared to react with an epitope at or near the same epitope recognized by C172. We developed a two-step ELISA with excellent sensitivity and specificity for rotavirus detection by utilizing C127 and/or C214 as a capture antibody and rabbit anti-rotavirus conjugated with horseradish peroxidase as a probe. Also, when both monoclonal C127 capture antibody and polyclonal rabbit anti-rotavirus-HRP were incubated with rotavirus simultaneously in a one-step assay, equivalent sensitivity and specificity were observed. The data show that these generated anti-rotavirus antibodies can be utilized effectively as reagents for the detection of human rotaviruses in stool specimens.