The development of nanor eactor for high sensitive chemi luminescence analysis by using liposomes

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Horseradish peroxidase (HRP) -encapsulated liposomes were prepared by freeze-thawing method. The number of HRP molecules trapped in liposomes was three times greater than that prepared by extrusion method HRP trapped in liposomes was directly detected by using luminol chemiluminescence (CL) with H_2O_2 without lysis of liposomes. The CL measurement conditions in both a lipid-free bulk solution and in liposomes were optimized in the concentrations of luminol and H_2O_2 by measuring the CL response curves, in which only one peak appeared and the CL intensity was maximal. The CL intensity observed in HRP-catalysed luminol CL in liposomes was a factor of seven greater than that observed in a lipid-free bulk solution. The detection limit in the direct detection of HRP encapsulated in liposomes was sensitive by a factor of 3 compared with that in HRP-catalysed luminol CL in a lipid-free bulk solution. In addition, HRP-trapped liposomes were combined to antibody in order to apply HRP-trapped liposomes to the marker in immunoassay. The CL intensity observed in antibody combined HRP-trapped liposomes was greater 125 times that in antibody combined HRP by avidin-biotin bond.