Single-molecule enzyme-linked immunosorbent assay detects serum proteins at subfemtomolar concentrations

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Rissin et al. increase the sensitivity of sandwich ELISA by segregating beads bearing a single enzyme-labeled immunoconjugate into femtoliter-volume reaction chambers. As the small volume of each well permits detection of extremely low levels of fluorescence, protein abundance is determined by counting the number of fluorescent wells as a percentage of the number of wells containing beads.

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