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

By David M. Rissin, C. Kan, Todd G. Campbell, S. Howes, David R. Fournier, Linan Song, T. Piech, Purvish P. Patel, Lei Chang, Andrew J. Rivnak and 5 more. Nature Biotechnology, 2010

1,942 citations recorded by the source corpus.

The ability to detect single protein molecules in blood could accelerate the discovery and use of more sensitive diagnostic biomarkers. To detect low-abundance proteins in blood, we captured them on microscopic beads decorated with specific antibodies and then labeled the immunocomplexes (one or zero labeled target protein molecules per bead) with an enzymatic reporter capable of generating a fluorescent product. After isolating the beads in 50-fl reaction chambers designed to hold only a single bead, we used fluorescence imaging to detect single protein molecules. Our single-molecule enzyme-linked immunosorbent assay (digital ELISA) approach detected as few as ∼10–20 enzyme-labeled complexes in 100 μl of sample (∼10−19 M) and routinely allowed detection of clinically relevant proteins in serum at concentrations (<10−15 M) much lower than conventional ELISA. Digital ELISA detected prostate-specific antigen (PSA) in sera from patients who had undergone radical prostatectomy at concentrations as low as 14 fg/ml (0.4 fM).

Subfields: Electrical and Electronic Engineering, Molecular Biology

View Paper: https://www.semanticscholar.org/paper/15d66460b7561dae20aac2a0dbc39b230464a7e2

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Citations in the corpus

• Proteomics: Technologies and Their Applications. (2017, methodology): “This assay was capable to detect 1,1-Dichloro-2,2-bis (p-chlorophenyl) ethylene (p,p′-DDE); a metabolite of insecticide and persistent organic pollutant that accumulates in food chain and environment (50).”
• Small but Perfectly Formed? Successes, Challenges, and Opportunities for Microfluidics in the Chemical and Biological Sciences (2017, methodology): “Reprinted by permission from Macmillan Publishers Ltd: Nature Biotechnology (Rissin et al.(104)), copyright 2010.”