schliessen

Filtern

 

Bibliotheken

Fluorescence anisotropy reduction of allosteric aptamer for sensitive and specific protein signaling

Real time protein signaling in a complex medium may provide a promising way for high-throughput protein analysis, but it is largely unmet due to the challenge of signal transduction and the interferences of nonspecific binding and high background. Our recent work indicates that a fluorescent aptamer... Full description

Journal Title: Analytical chemistry 03 April 2012, Vol.84(7), pp.3070-4
Main Author: Zhang, Dapeng
Other Authors: Zhao, Qiang , Zhao, Bailin , Wang, Hailin
Format: Electronic Article Electronic Article
Language: English
Subjects:
ID: E-ISSN: 1520-6882 ; PMID: 22455347 Version:1 ; DOI: 10.1021/ac3004133
Link: http://pubmed.gov/22455347
Zum Text:
SendSend as email Add to Book BagAdd to Book Bag
Staff View
recordid: medline22455347
title: Fluorescence anisotropy reduction of allosteric aptamer for sensitive and specific protein signaling
format: Article
creator:
  • Zhang, Dapeng
  • Zhao, Qiang
  • Zhao, Bailin
  • Wang, Hailin
subjects:
  • Aptamers, Nucleotide -- Metabolism
  • Biological Assay -- Methods
  • Fluorescence Polarization -- Methods
  • Thrombin -- Metabolism
ispartof: Analytical chemistry, 03 April 2012, Vol.84(7), pp.3070-4
description: Real time protein signaling in a complex medium may provide a promising way for high-throughput protein analysis, but it is largely unmet due to the challenge of signal transduction and the interferences of nonspecific binding and high background. Our recent work indicates that a fluorescent aptamer can display a protein binding-induced reduction of fluorescence anisotropy (FA) (Zhang, D.; Lu, M.; Wang, H. J. Am. Chem. Soc. 2011, 133, 9188-9191), which is exclusively different from a traditionally simplified concept hinting a molecular size increase-induced FA increase. Inspired by this unexpected observation, we describe a novel FA reduction approach for protein signaling. The feasibility of this approach is demonstrated through the assays of a blood protein human α-thrombin and an oncoprotein human platelet-derived growth factor B-chain (PDGF-BB) using two screened fluorescent aptamers, respectively. By the developed FA reduction method, the spiked human α-thrombin in diluted serum can be detected at the concentration as low as 250 pM. In contrast, in a traditional molecular size-dependent FA assay, the thrombin spiked in diluted serum cannot induce reliable FA change even at a 256-fold higher concentration (64 nM). The results clearly show that the FA reduction approach has a dramatically enhanced specificity against target protein and high sensitivity in complex medium and is applicable to the no-separation based detection of proteins in biological matrixes.
language: eng
source:
identifier: E-ISSN: 1520-6882 ; PMID: 22455347 Version:1 ; DOI: 10.1021/ac3004133
fulltext: no_fulltext
issn:
  • 15206882
  • 1520-6882
url: Link


@attributes
ID1546228048
RANK0.07
NO1
SEARCH_ENGINEprimo_central_multiple_fe
SEARCH_ENGINE_TYPEPrimo Central Search Engine
LOCALfalse
PrimoNMBib
record
control
sourcerecordid22455347
sourceidmedline
recordidTN_medline22455347
sourceformatXML
sourcesystemOther
pqid964196319
galeid305486432
display
typearticle
titleFluorescence anisotropy reduction of allosteric aptamer for sensitive and specific protein signaling
creatorZhang, Dapeng ; Zhao, Qiang ; Zhao, Bailin ; Wang, Hailin
ispartofAnalytical chemistry, 03 April 2012, Vol.84(7), pp.3070-4
identifier
subjectAptamers, Nucleotide -- Metabolism ; Biological Assay -- Methods ; Fluorescence Polarization -- Methods ; Thrombin -- Metabolism
descriptionReal time protein signaling in a complex medium may provide a promising way for high-throughput protein analysis, but it is largely unmet due to the challenge of signal transduction and the interferences of nonspecific binding and high background. Our recent work indicates that a fluorescent aptamer can display a protein binding-induced reduction of fluorescence anisotropy (FA) (Zhang, D.; Lu, M.; Wang, H. J. Am. Chem. Soc. 2011, 133, 9188-9191), which is exclusively different from a traditionally simplified concept hinting a molecular size increase-induced FA increase. Inspired by this unexpected observation, we describe a novel FA reduction approach for protein signaling. The feasibility of this approach is demonstrated through the assays of a blood protein human α-thrombin and an oncoprotein human platelet-derived growth factor B-chain (PDGF-BB) using two screened fluorescent aptamers, respectively. By the developed FA reduction method, the spiked human α-thrombin in diluted serum can be detected at the concentration as low as 250 pM. In contrast, in a traditional molecular size-dependent FA assay, the thrombin spiked in diluted serum cannot induce reliable FA change even at a 256-fold higher concentration (64 nM). The results clearly show that the FA reduction approach has a dramatically enhanced specificity against target protein and high sensitivity in complex medium and is applicable to the no-separation based detection of proteins in biological matrixes.
languageeng
source
version5
lds50peer_reviewed
links
openurl$$Topenurl_article
backlink$$Uhttp://pubmed.gov/22455347$$EView_this_record_in_MEDLINE/PubMed
openurlfulltext$$Topenurlfull_article
addlink$$Uhttp://exlibris-pub.s3.amazonaws.com/aboutMedline.html$$EView_the_MEDLINE/PubMed_Copyright_Statement
search
creatorcontrib
0Zhang, Dapeng
1Zhao, Qiang
2Zhao, Bailin
3Wang, Hailin
titleFluorescence anisotropy reduction of allosteric aptamer for sensitive and specific protein signaling
descriptionReal time protein signaling in a complex medium may provide a promising way for high-throughput protein analysis, but it is largely unmet due to the challenge of signal transduction and the interferences of nonspecific binding and high background. Our recent work indicates that a fluorescent aptamer can display a protein binding-induced reduction of fluorescence anisotropy (FA) (Zhang, D.; Lu, M.; Wang, H. J. Am. Chem. Soc. 2011, 133, 9188-9191), which is exclusively different from a traditionally simplified concept hinting a molecular size increase-induced FA increase. Inspired by this unexpected observation, we describe a novel FA reduction approach for protein signaling. The feasibility of this approach is demonstrated through the assays of a blood protein human α-thrombin and an oncoprotein human platelet-derived growth factor B-chain (PDGF-BB) using two screened fluorescent aptamers, respectively. By the developed FA reduction method, the spiked human α-thrombin in diluted serum can be detected at the concentration as low as 250 pM. In contrast, in a traditional molecular size-dependent FA assay, the thrombin spiked in diluted serum cannot induce reliable FA change even at a 256-fold higher concentration (64 nM). The results clearly show that the FA reduction approach has a dramatically enhanced specificity against target protein and high sensitivity in complex medium and is applicable to the no-separation based detection of proteins in biological matrixes.
subject
0Aptamers, Nucleotide -- Metabolism
1Biological Assay -- Methods
2Fluorescence Polarization -- Methods
3Thrombin -- Metabolism
general
022455347
1English
2MEDLINE/PubMed (U.S. National Library of Medicine)
310.1021/ac3004133
4MEDLINE/PubMed (NLM)
sourceidmedline
recordidmedline22455347
issn
015206882
11520-6882
rsrctypearticle
creationdate2012
addtitleAnalytical chemistry
searchscope
0medline
1nlm_medline
2MEDLINE
scope
0medline
1nlm_medline
2MEDLINE
lsr4120120403
citationpf 3070 vol 84 issue 7
startdate20120403
enddate20120403
lsr30VSR-Enriched:[galeid, pages, issn, pqid]
sort
titleFluorescence anisotropy reduction of allosteric aptamer for sensitive and specific protein signaling
authorZhang, Dapeng ; Zhao, Qiang ; Zhao, Bailin ; Wang, Hailin
creationdate20120403
lso0120120403
facets
frbrgroupid8526693266657746688
frbrtype5
newrecords20190701
languageeng
creationdate2012
topic
0Aptamers, Nucleotide–Metabolism
1Biological Assay–Methods
2Fluorescence Polarization–Methods
3Thrombin–Metabolism
collectionMEDLINE/PubMed (NLM)
prefilterarticles
rsrctypearticles
creatorcontrib
0Zhang, Dapeng
1Zhao, Qiang
2Zhao, Bailin
3Wang, Hailin
jtitleAnalytical Chemistry
toplevelpeer_reviewed
delivery
delcategoryRemote Search Resource
fulltextno_fulltext
addata
aulast
0Zhang
1Zhao
2Wang
aufirst
0Dapeng
1Qiang
2Bailin
3Hailin
au
0Zhang, Dapeng
1Zhao, Qiang
2Zhao, Bailin
3Wang, Hailin
atitleFluorescence anisotropy reduction of allosteric aptamer for sensitive and specific protein signaling
jtitleAnalytical chemistry
risdate20120403
volume84
issue7
spage3070
pages3070-3074
eissn1520-6882
formatjournal
genrearticle
ristypeJOUR
abstractReal time protein signaling in a complex medium may provide a promising way for high-throughput protein analysis, but it is largely unmet due to the challenge of signal transduction and the interferences of nonspecific binding and high background. Our recent work indicates that a fluorescent aptamer can display a protein binding-induced reduction of fluorescence anisotropy (FA) (Zhang, D.; Lu, M.; Wang, H. J. Am. Chem. Soc. 2011, 133, 9188-9191), which is exclusively different from a traditionally simplified concept hinting a molecular size increase-induced FA increase. Inspired by this unexpected observation, we describe a novel FA reduction approach for protein signaling. The feasibility of this approach is demonstrated through the assays of a blood protein human α-thrombin and an oncoprotein human platelet-derived growth factor B-chain (PDGF-BB) using two screened fluorescent aptamers, respectively. By the developed FA reduction method, the spiked human α-thrombin in diluted serum can be detected at the concentration as low as 250 pM. In contrast, in a traditional molecular size-dependent FA assay, the thrombin spiked in diluted serum cannot induce reliable FA change even at a 256-fold higher concentration (64 nM). The results clearly show that the FA reduction approach has a dramatically enhanced specificity against target protein and high sensitivity in complex medium and is applicable to the no-separation based detection of proteins in biological matrixes.
doi10.1021/ac3004133
pmid22455347
issn00032700
oafree_for_read
date2012-04-03