SINGLE MOLECULE DETECTION OF ALKALINE PHOSPHATASE ENZYME

摘要

Enzymatic triggerable 1,2-dioxetanes are superior in immunoassays and other related applications compared to peroxidase substrates such as luminol and others. Stabilized 1,2-dioxetane substrates provide high signal, low background, wide dynamic range, rapid results and excellent reproducibility. These 1,2-dioxetanes provide substrates which are highly sensitive but can detect an enzyme concentration up to 10~(-21)M (6x10~2 molecules of alkaline phosphatase) in solution as well as on a membrane. Now we wish to report on the detection of a single molecule of alkaline phosphatase, in less than ten minutes, by using a combination of a stabilized 1,2-dioxetanes, an enhancer and an enzyme diluent.