Future immunoassays and nucleic acid hybridization assays will be performed in miniaturized formats that utilize microchips or microparticles. This will require a sensitive detection technology that allows spatial resolution. By using fluorescent europium chelates and time-resolved microfluorometry, one can detect 11 000 europium molecules on individual microparticles. In a miniaturized noncompetitive immunoassay of prostate-specific antigen (PSA), we quantitatively detected 5 ng/L (0.05 amol per particle) of the analyte on an individual microparticle with excellent precision over the whole measurement range (CV <10%). Using a hybridization assay, we also could detect the ΔF508 mutation for cystic fibrosis on individual microparticles. Consequently, fluorescent lanthanide chelate labels and time-resolved microfluorometry qualify as the next generation of technology in this field.
Three monoclonal antibodies, termed 4E10, lE11:10, and 2D9:1, were generated against rubella virus. lmmunoblot analysis with purified authentic rubella virus or recombinant baculovirus-expressed rubella virus structural proteins El, E2, and C demonstrated that they were directed against the El envelope glycoprotein of the rubella virus particle. By using the yeast Ty virus-like particle system, it was possible to map the binding site of lE11:10 within amino acids 236 to 286 of the El protein and the binding sites of 2D9:1 and 4E10 outside this region. Immunoaffinity purification with these monoclonal antibodies made it evident that they are useful for obtaining large quantities of pure baculovirus-expressed rubella virus envelope protein El. The diagnostic potential of this immunoaffinity-purified recombinant rubella virus El protein compared with that of authentic rubella virus is demonstrated.
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