当前位置： SCI文献检索 > NATURE BIOTECHNOLOGY期刊下所有文献 > Single-molecule enzyme-linked immunosorbent assay detects serum proteins at subfemtomolar concentrations.

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

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_name

Nat Biotechnol

journal_title

Nature biotechnology

authors

Rissin DM,Kan CW,Campbell TG,Howes SC,Fournier DR,Song L,Piech T,Patel PP,Chang L,Rivnak AJ,Ferrell EP,Randall JD,Provuncher GK,Walt DR,Duffy DC

doi

10.1038/nbt.1641

subject

Has Abstract

pub_date

2010-06-01 00:00:00

pages

595-9

issue

6

eissn

1087-0156

issn

1546-1696

pii

nbt.1641

journal_volume

28

pub_type

杂志文章

相关文献

NATURE BIOTECHNOLOGY文献大全
  • A functional screen in yeast for regulators and antagonizers of heterologous protein tyrosine kinases.

    abstract：:Tyrosine phosphorylation exerts a pivotal role in cell regulation processes of higher eukaryotes. Tight control of the activity of protein tyrosine kinases is crucial for ordered phosphorylation to occur. We have developed a functional screen for tyrosine kinase regulators using c-Src, the first cellular protein tyros...

    journal_title：Nature biotechnology

    pub_type： 杂志文章

    doi：10.1038/nbt0596-600

    authors： Superti-Furga G,Jönsson K,Courtneidge SA

    更新日期：1996-05-01 00:00:00

  • Microarray reality checks in the context of a complex disease.

    abstract：:A problem in analyzing microarray-based gene expression data is the separation of genes causally involved in a disease from innocent bystander genes, whose expression levels have been secondarily altered by primary changes elsewhere. To investigate this issue systematically in the context of a class of complex human d...

    journal_title：Nature biotechnology

    pub_type： 杂志文章

    doi：10.1038/nbt965

    authors： Miklos GL,Maleszka R

    更新日期：2004-05-01 00:00:00

  • Flexibility--the guiding principle for antibody manufacturing.

    abstract：:Ensuring adequate manufacturing capacity for recombinant and monoclonal antibodies is one of the greatest challenges facing the biotech industry. The following factors should be taken into account when designing product development and production strategies. ...

    journal_title：Nature biotechnology

    pub_type： 杂志文章

    doi：10.1038/nbt0905-1054

    authors： Carson KL

    更新日期：2005-09-01 00:00:00

  • Cell penetrating PNA constructs regulate galanin receptor levels and modify pain transmission in vivo.

    abstract：:Peptide nucleic acids (PNAs) form stable and tight complexes with complementary DNA and/or RNA and would be promising antisense reagents if their cellular delivery could be improved. We show that a 21-mer PNA, complementary to the human galanin receptor type 1 mRNA, coupled to the cellular transporter peptides, transp...

    journal_title：Nature biotechnology

    pub_type： 杂志文章

    doi：10.1038/nbt0998-857

    authors： Pooga M,Soomets U,Hällbrink M,Valkna A,Saar K,Rezaei K,Kahl U,Hao JX,Xu XJ,Wiesenfeld-Hallin Z,Hökfelt T,Bartfai T,Langel U

    更新日期：1998-09-01 00:00:00

  • Prevention of systemic lupus erythematosus in MRL/lpr mice by administration of an immunoglobulin-binding peptide.

    abstract：:Systemic lupus erythematosus (SLE) is a multisystem chronic inflammatory disease of unknown etiology that affects many organs, including the kidney. The presence of multiple autoantibodies and other immunological abnormalities point to basic defects in immunoregulatory controls that normally maintain self-tolerance. T...

    journal_title：Nature biotechnology

    pub_type： 杂志文章

    doi：10.1038/77296

    authors： Marino M,Ruvo M,De Falco S,Fassina G

    更新日期：2000-07-01 00:00:00

  • An integrated mass spectrometric and computational framework for the analysis of protein interaction networks.

    abstract：:Biological systems are controlled by protein complexes that associate into dynamic protein interaction networks. We describe a strategy that analyzes protein complexes through the integration of label-free, quantitative mass spectrometry and computational analysis. By evaluating peptide intensity profiles throughout t...

    journal_title：Nature biotechnology

    pub_type： 杂志文章

    doi：10.1038/nbt1289

    authors： Rinner O,Mueller LN,Hubálek M,Müller M,Gstaiger M,Aebersold R

    更新日期：2007-03-01 00:00:00

  • Design of stapled antimicrobial peptides that are stable, nontoxic and kill antibiotic-resistant bacteria in mice.

    abstract：:The clinical translation of cationic α-helical antimicrobial peptides (AMPs) has been hindered by structural instability, proteolytic degradation and in vivo toxicity from nonspecific membrane lysis. Although analyses of hydrophobic content and charge distribution have informed the design of synthetic AMPs with increa...

    journal_title：Nature biotechnology

    pub_type： 杂志文章

    doi：10.1038/s41587-019-0222-z

    authors： Mourtada R,Herce HD,Yin DJ,Moroco JA,Wales TE,Engen JR,Walensky LD

    更新日期：2019-10-01 00:00:00

  • A human beta-cell line for transplantation therapy to control type 1 diabetes.

    abstract：:A human pancreatic beta-cell line that is functionally equivalent to primary beta-cells has not been available. We established a reversibly immortalized human beta-cell clone (NAKT-15) by transfection of primary human beta-cells with a retroviral vector containing simian virus 40 large T-antigen (SV40T) and human telo...

    journal_title：Nature biotechnology

    pub_type： 杂志文章

    doi：10.1038/nbt1145

    authors： Narushima M,Kobayashi N,Okitsu T,Tanaka Y,Li SA,Chen Y,Miki A,Tanaka K,Nakaji S,Takei K,Gutierrez AS,Rivas-Carrillo JD,Navarro-Alvarez N,Jun HS,Westerman KA,Noguchi H,Lakey JR,Leboulch P,Tanaka N,Yoon JW

    更新日期：2005-10-01 00:00:00

  • The molecular properties and applications of Anthozoa fluorescent proteins and chromoproteins.

    abstract：:The green fluorescent protein (GFP) from the jellyfish Aequorea victoria and its fluorescent homologs from Anthozoa corals have become invaluable tools for in vivo imaging of cells and tissues. Despite spectral and chromophore diversity, about 100 cloned members of the GFP-like protein family possess common structural...

    journal_title：Nature biotechnology

    pub_type： 杂志文章,评审

    doi：10.1038/nbt943

    authors： Verkhusha VV,Lukyanov KA

    更新日期：2004-03-01 00:00:00

  • Selection for a periplasmic factor improving phage display and functional periplasmic expression.

    abstract：:The efficiency of both phage display in Escherichia coli and periplasmic expression of recombinant proteins may be limited by the same periplasmic folding steps. To search for E. coli factors that improve the efficiency of both procedures, a library of E. coli proteins was coexpressed in a phagemid vector that contain...

    journal_title：Nature biotechnology

    pub_type： 杂志文章

    doi：10.1038/nbt0498-376

    authors： Bothmann H,Plückthun A

    更新日期：1998-04-01 00:00:00

  • DNA variation and the future of human genetics.

    abstract：:The use of DNA variants in the mapping of the human genome and in the positional cloning of monogenic disease genes is well established. Determining the genetic bases of the more common "multifactorial" diseases, however, presents a major challenge. The genetics of these diseases are complicated by the interplay betwe...

    journal_title：Nature biotechnology

    pub_type： 杂志文章,评审

    doi：10.1038/nbt0198-33

    authors： Schafer AJ,Hawkins JR

    更新日期：1998-01-01 00:00:00

  • Milking the genome for profit.

    abstract：:Despite recent accomplishments, significant challenges remain before cloning can become a routine part of transgenic animal production. ...

    journal_title：Nature biotechnology

    pub_type： 杂志文章

    doi：10.1038/80231

    authors： Dove A

    更新日期：2000-10-01 00:00:00

  • Synthetic CD4 exocyclics inhibit binding of human immunodeficiency virus type 1 envelope to CD4 and virus replication in T lymphocytes.

    abstract：:CD4 functions as a major T-cell surface receptor for human immunodeficiency virus by binding the human immunodeficiency virus type 1 (HIV-1) envelope protein gp120 with relatively high affinity. We have developed constrained aromatically modified analogs of the secondary structures of the first domain of CD4 in order ...

    journal_title：Nature biotechnology

    pub_type： 杂志文章

    doi：10.1038/nbt0297-150

    authors： Zhang X,Gaubin M,Briant L,Srikantan V,Murali R,Saragovi U,Weiner D,Devaux C,Autiero M,Piatier-Tonneau D,Greene MI

    更新日期：1997-02-01 00:00:00

  • Pulsed electric fields for selection of hematopoietic cells and depletion of tumor cell contaminants.

    abstract：:Purging of tumor cells and selection of stem cells are key technologies for enabling stem cell transplantation and stem cell gene therapy. Here we report a strategy for cell selection based on physical properties of the cells. Exposing cells to an external pulsed electric field (PEF) increases the natural potential di...

    journal_title：Nature biotechnology

    pub_type： 杂志文章

    doi：10.1038/78504

    authors： Eppich HM,Foxall R,Gaynor K,Dombkowski D,Miura N,Cheng T,Silva-Arrieta S,Evans RH,Mangano JA,Preffer FI,Scadden DT

    更新日期：2000-08-01 00:00:00

  • Author Correction: Plants with genetically encoded autoluminescence.

    abstract：:An amendment to this paper has been published and can be accessed via a link at the top of the paper. ...

    journal_title：Nature biotechnology

    pub_type： 已发布勘误

    doi：10.1038/s41587-020-0578-0

    authors： Mitiouchkina T,Mishin AS,Somermeyer LG,Markina NM,Chepurnyh TV,Guglya EB,Karataeva TA,Palkina KA,Shakhova ES,Fakhranurova LI,Chekova SV,Tsarkova AS,Golubev YV,Negrebetsky VV,Dolgushin SA,Shalaev PV,Shlykov D,Melnik OA,

    更新日期：2020-08-01 00:00:00

  • Site-specific genomic integration produces therapeutic Factor IX levels in mice.

    abstract：:We used the integrase from phage phiC31 to integrate the human Factor IX (hFIX) gene permanently into specific sites in the mouse genome. A plasmid containing attB and an expression cassette for hFIX was delivered to the livers of mice by using high-pressure tail vein injection. When an integrase expression plasmid wa...

    journal_title：Nature biotechnology

    pub_type： 杂志文章

    doi：10.1038/nbt753

    authors： Olivares EC,Hollis RP,Chalberg TW,Meuse L,Kay MA,Calos MP

    更新日期：2002-11-01 00:00:00

  • Patent protection for protein structure analysis.

    abstract：:Three national patent offices have consulted on patents that cover protein three-dimensional structural data and pharmacophores, with significant implications for the biotechnology industry. ...

    journal_title：Nature biotechnology

    pub_type： 杂志文章

    doi：10.1038/nbt0104-109

    authors： Shimbo I,Nakajima R,Yokoyama S,Sumikura K

    更新日期：2004-01-01 00:00:00

  • RNA interference against viruses: strike and counterstrike.

    abstract：:RNA interference (RNAi) is a conserved sequence-specific, gene-silencing mechanism that is induced by double-stranded RNA. RNAi holds great promise as a novel nucleic acid-based therapeutic against a wide variety of diseases, including cancer, infectious diseases and genetic disorders. Antiviral RNAi strategies have r...

    journal_title：Nature biotechnology

    pub_type： 杂志文章,评审

    doi：10.1038/nbt1369

    authors： Haasnoot J,Westerhout EM,Berkhout B

    更新日期：2007-12-01 00:00:00

  • The complete genome sequence of the meat-borne lactic acid bacterium Lactobacillus sakei 23K.

    abstract：:Lactobacillus sakei is a psychotrophic lactic acid bacterium found naturally on fresh meat and fish. This microorganism is widely used in the manufacture of fermented meats and has biotechnological potential in biopreservation and food safety. We have explored the 1,884,661-base-pair (bp) circular chromosome of strain...

    journal_title：Nature biotechnology

    pub_type： 杂志文章

    doi：10.1038/nbt1160

    authors： Chaillou S,Champomier-Vergès MC,Cornet M,Crutz-Le Coq AM,Dudez AM,Martin V,Beaufils S,Darbon-Rongère E,Bossy R,Loux V,Zagorec M

    更新日期：2005-12-01 00:00:00

  • Large-scale detection of antigen-specific T cells using peptide-MHC-I multimers labeled with DNA barcodes.

    abstract：:Identification of the peptides recognized by individual T cells is important for understanding and treating immune-related diseases. Current cytometry-based approaches are limited to the simultaneous screening of 10-100 distinct T-cell specificities in one sample. Here we use peptide-major histocompatibility complex (...

    journal_title：Nature biotechnology

    pub_type： 杂志文章

    doi：10.1038/nbt.3662

    authors： Bentzen AK,Marquard AM,Lyngaa R,Saini SK,Ramskov S,Donia M,Such L,Furness AJ,McGranahan N,Rosenthal R,Straten PT,Szallasi Z,Svane IM,Swanton C,Quezada SA,Jakobsen SN,Eklund AC,Hadrup SR

    更新日期：2016-10-01 00:00:00

  • Epigenome-wide association data implicate DNA methylation as an intermediary of genetic risk in rheumatoid arthritis.

    abstract：:Epigenetic mechanisms integrate genetic and environmental causes of disease, but comprehensive genome-wide analyses of epigenetic modifications have not yet demonstrated robust association with common diseases. Using Illumina HumanMethylation450 arrays on 354 anti-citrullinated protein antibody-associated rheumatoid a...

    journal_title：Nature biotechnology

    pub_type： 杂志文章

    doi：10.1038/nbt.2487

    authors： Liu Y,Aryee MJ,Padyukov L,Fallin MD,Hesselberg E,Runarsson A,Reinius L,Acevedo N,Taub M,Ronninger M,Shchetynsky K,Scheynius A,Kere J,Alfredsson L,Klareskog L,Ekström TJ,Feinberg AP

    更新日期：2013-02-01 00:00:00

  • Isotopic labeling of terminal amines in complex samples identifies protein N-termini and protease cleavage products.

    abstract：:Effective proteome-wide strategies that distinguish the N-termini of proteins from the N-termini of their protease cleavage products would accelerate identification of the substrates of proteases with broad or unknown specificity. Our approach, named terminal amine isotopic labeling of substrates (TAILS), addresses th...

    journal_title：Nature biotechnology

    pub_type： 杂志文章

    doi：10.1038/nbt.1611

    authors： Kleifeld O,Doucet A,auf dem Keller U,Prudova A,Schilling O,Kainthan RK,Starr AE,Foster LJ,Kizhakkedathu JN,Overall CM

    更新日期：2010-03-01 00:00:00

  • Alteration of Cre recombinase site specificity by substrate-linked protein evolution.

    abstract：:Directed molecular evolution was applied to generate Cre recombinase variants that recognize a new DNA target sequence. Cre was adapted in a three-stage strategy to evolve recombinases to specifically recombine the new site. This complex multicycle task was made feasible by an improved directed-evolution procedure tha...

    journal_title：Nature biotechnology

    pub_type： 杂志文章

    doi：10.1038/nbt1101-1047

    authors： Buchholz F,Stewart AF

    更新日期：2001-11-01 00:00:00

  • Canola engineered with a microalgal polyketide synthase-like system produces oil enriched in docosahexaenoic acid.

    abstract：:Dietary omega-3 long-chain polyunsaturated fatty acids (LC-PUFAs), docosahexaenoic acid (DHA, C22:6) and eicosapentaenoic acid (EPA, C20:5) are usually derived from marine fish. Although production of both EPA and DHA has been engineered into land plants, including Arabidopsis, Camelina sativa and Brassica juncea, nei...

    journal_title：Nature biotechnology

    pub_type： 杂志文章

    doi：10.1038/nbt.3585

    authors： Walsh TA,Bevan SA,Gachotte DJ,Larsen CM,Moskal WA,Merlo PA,Sidorenko LV,Hampton RE,Stoltz V,Pareddy D,Anthony GI,Bhaskar PB,Marri PR,Clark LM,Chen W,Adu-Peasah PS,Wensing ST,Zirkle R,Metz JG

    更新日期：2016-08-01 00:00:00

  • Microbial synthesis of medium-chain chemicals from renewables.

    abstract：:Linear, medium-chain (C8-C12) hydrocarbons are important components of fuels as well as commodity and specialty chemicals. As industrial microbes do not contain pathways to produce medium-chain chemicals, approaches such as overexpression of endogenous enzymes or deletion of competing pathways are not available to the...

    journal_title：Nature biotechnology

    pub_type： 杂志文章,评审

    doi：10.1038/nbt.4022

    authors： Sarria S,Kruyer NS,Peralta-Yahya P

    更新日期：2017-12-01 00:00:00

  • Short hairpin RNA-expressing bacteria elicit RNA interference in mammals.

    abstract：:RNA-interference (RNAi) is a potent mechanism, conserved from plants to humans for specific silencing of genes, which holds promise for functional genomics and gene-targeted therapies. Here we show that bacteria engineered to produce a short hairpin RNA (shRNA) targeting a mammalian gene induce trans-kingdom RNAi in v...

    journal_title：Nature biotechnology

    pub_type： 杂志文章

    doi：10.1038/nbt1211

    authors： Xiang S,Fruehauf J,Li CJ

    更新日期：2006-06-01 00:00:00

  • Engineered off-the-shelf therapeutic T cells resist host immune rejection.

    abstract：:Engineered T cells are effective therapies against a range of malignancies, but current approaches rely on autologous T cells, which are difficult and expensive to manufacture. Efforts to develop potent allogeneic T cells that are not rejected by the recipient's immune system require abrogating both T- and natural kil...

    journal_title：Nature biotechnology

    pub_type： 杂志文章

    doi：10.1038/s41587-020-0601-5

    authors： Mo F,Watanabe N,McKenna MK,Hicks MJ,Srinivasan M,Gomes-Silva D,Atilla E,Smith T,Ataca Atilla P,Ma R,Quach D,Heslop HE,Brenner MK,Mamonkin M

    更新日期：2021-01-01 00:00:00

  • Adeno-associated virus serotype 8 efficiently delivers genes to muscle and heart.

    abstract：:Systemic gene delivery into muscle has been a major challenge for muscular dystrophy gene therapy, with capillary blood vessels posing the principle barrier and limiting vector dissemination. Previous efforts to deliver genes into multiple muscles have relied on isolated vessel perfusion or pharmacological interventio...

    journal_title：Nature biotechnology

    pub_type： 杂志文章

    doi：10.1038/nbt1073

    authors： Wang Z,Zhu T,Qiao C,Zhou L,Wang B,Zhang J,Chen C,Li J,Xiao X

    更新日期：2005-03-01 00:00:00

  • MaxQuant enables high peptide identification rates, individualized p.p.b.-range mass accuracies and proteome-wide protein quantification.

    abstract：:Efficient analysis of very large amounts of raw data for peptide identification and protein quantification is a principal challenge in mass spectrometry (MS)-based proteomics. Here we describe MaxQuant, an integrated suite of algorithms specifically developed for high-resolution, quantitative MS data. Using correlatio...

    journal_title：Nature biotechnology

    pub_type： 杂志文章

    doi：10.1038/nbt.1511

    authors： Cox J,Mann M

    更新日期：2008-12-01 00:00:00

  • Donor cell type can influence the epigenome and differentiation potential of human induced pluripotent stem cells.

    abstract：:We compared bona fide human induced pluripotent stem cells (iPSCs) derived from umbilical cord blood (CB) cells and neonatal keratinocytes (K). As a consequence of both incomplete erasure of tissue-specific methylation and aberrant de novo methylation, CB-iPSCs and K-iPSCs were distinct in genome-wide DNA methylation ...

    journal_title：Nature biotechnology

    pub_type： 杂志文章

    doi：10.1038/nbt.2052

    authors： Kim K,Zhao R,Doi A,Ng K,Unternaehrer J,Cahan P,Huo H,Loh YH,Aryee MJ,Lensch MW,Li H,Collins JJ,Feinberg AP,Daley GQ

    更新日期：2011-11-27 00:00:00