{"database":"biostudies-literature","file_versions":[],"scores":null,"additional":{"submitter":["Rissin DM"],"funding":["NCI NIH HHS"],"pagination":["595-9"],"full_dataset_link":["https://www.ebi.ac.uk/biostudies/studies/S-EPMC2919230"],"repository":["biostudies-literature"],"omics_type":["Unknown"],"volume":["28(6)"],"pubmed_abstract":["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 approximately 10-20 enzyme-labeled complexes in 100 microl of sample (approximately 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)."],"journal":["Nature biotechnology"],"pubmed_title":["Single-molecule enzyme-linked immunosorbent assay detects serum proteins at subfemtomolar concentrations."],"pmcid":["PMC2919230"],"funding_grant_id":["R43CA133987","R43 CA133987-01","R43 CA133987-02","R43 CA133987","R44 CA133987"],"pubmed_authors":["Chang L","Rissin DM","Fournier DR","Provuncher GK","Howes SC","Song L","Randall JD","Rivnak AJ","Walt DR","Kan CW","Piech T","Campbell TG","Patel PP","Ferrell EP","Duffy DC"],"additional_accession":[]},"is_claimable":false,"name":"Single-molecule enzyme-linked immunosorbent assay detects serum proteins at subfemtomolar concentrations.","description":"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 approximately 10-20 enzyme-labeled complexes in 100 microl of sample (approximately 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).","dates":{"release":"2010-01-01T00:00:00Z","publication":"2010 Jun","modification":"2025-04-04T11:32:45.458Z","creation":"2019-03-27T00:33:08Z"},"accession":"S-EPMC2919230","cross_references":{"pubmed":["20495550"],"doi":["10.1038/nbt.1641"]}}