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Single Molecule Array (SiMoA™) technology is based upon the isolation of individual immunocomplexes on beads using standard ELISA reagents.
Beads with or without labeled immunocomplexes are loaded into arrays containing femtoliter-sized wells, and the arrays are sealed in the presence of the enzyme substrate and fluorescently imaged. Fluorescent product molecules of the enzyme-substrate reaction are confined in a 50 femtoliter volume, giving rise to a local high concentration of product within a matter of a few seconds that can be easily detected.
By utilizing high density arrays containing 50,000-500,000 wells, hundreds to thousands of individual immunocomplexes can be detected simultaneously, permitting rapid data acquisition. Isolation of single immunocomplexes in this way gives rise to a dramatic increase in sensitivity to enzyme labels over bulk, ensemble detection methods employed in traditional immunoassay technology. SiMoA assays are easy to construct because they use the same reagents as traditional ELISA (antibodies, antigens and enzymes), but are routinely more than 1000-fold more sensitive than comparable ELISAs due to the single molecule nature of the technology. |
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