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Quantifying Nanomolar Protein Concentrations Using Designed DNA Carriers and Solid-State Nanopores

[Image: see text] Designed “DNA carriers” have been proposed as a new method for nanopore based specific protein detection. In this system, target protein molecules bind to a long DNA strand at a defined position creating a second level transient current drop against the background DNA translocation...

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Autores principales: Kong, Jinglin, Bell, Nicholas A. W., Keyser, Ulrich F.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: American Chemical Society 2016
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4901370/
https://www.ncbi.nlm.nih.gov/pubmed/27121643
http://dx.doi.org/10.1021/acs.nanolett.6b00627
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author Kong, Jinglin
Bell, Nicholas A. W.
Keyser, Ulrich F.
author_facet Kong, Jinglin
Bell, Nicholas A. W.
Keyser, Ulrich F.
author_sort Kong, Jinglin
collection PubMed
description [Image: see text] Designed “DNA carriers” have been proposed as a new method for nanopore based specific protein detection. In this system, target protein molecules bind to a long DNA strand at a defined position creating a second level transient current drop against the background DNA translocation. Here, we demonstrate the ability of this system to quantify protein concentrations in the nanomolar range. After incubation with target protein at different concentrations, the fraction of DNA translocations showing a secondary current spike allows for the quantification of the corresponding protein concentration. For our proof-of-principle experiments we use two standard binding systems, biotin–streptavidin and digoxigenin–antidigoxigenin, that allow for measurements of the concentration down to the low nanomolar range. The results demonstrate the potential for a novel quantitative and specific protein detection scheme using the DNA carrier method.
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spelling pubmed-49013702016-06-16 Quantifying Nanomolar Protein Concentrations Using Designed DNA Carriers and Solid-State Nanopores Kong, Jinglin Bell, Nicholas A. W. Keyser, Ulrich F. Nano Lett [Image: see text] Designed “DNA carriers” have been proposed as a new method for nanopore based specific protein detection. In this system, target protein molecules bind to a long DNA strand at a defined position creating a second level transient current drop against the background DNA translocation. Here, we demonstrate the ability of this system to quantify protein concentrations in the nanomolar range. After incubation with target protein at different concentrations, the fraction of DNA translocations showing a secondary current spike allows for the quantification of the corresponding protein concentration. For our proof-of-principle experiments we use two standard binding systems, biotin–streptavidin and digoxigenin–antidigoxigenin, that allow for measurements of the concentration down to the low nanomolar range. The results demonstrate the potential for a novel quantitative and specific protein detection scheme using the DNA carrier method. American Chemical Society 2016-04-28 2016-06-08 /pmc/articles/PMC4901370/ /pubmed/27121643 http://dx.doi.org/10.1021/acs.nanolett.6b00627 Text en Copyright © 2016 American Chemical Society This is an open access article published under a Creative Commons Attribution (CC-BY) License (http://pubs.acs.org/page/policy/authorchoice_ccby_termsofuse.html) , which permits unrestricted use, distribution and reproduction in any medium, provided the author and source are cited.
spellingShingle Kong, Jinglin
Bell, Nicholas A. W.
Keyser, Ulrich F.
Quantifying Nanomolar Protein Concentrations Using Designed DNA Carriers and Solid-State Nanopores
title Quantifying Nanomolar Protein Concentrations Using Designed DNA Carriers and Solid-State Nanopores
title_full Quantifying Nanomolar Protein Concentrations Using Designed DNA Carriers and Solid-State Nanopores
title_fullStr Quantifying Nanomolar Protein Concentrations Using Designed DNA Carriers and Solid-State Nanopores
title_full_unstemmed Quantifying Nanomolar Protein Concentrations Using Designed DNA Carriers and Solid-State Nanopores
title_short Quantifying Nanomolar Protein Concentrations Using Designed DNA Carriers and Solid-State Nanopores
title_sort quantifying nanomolar protein concentrations using designed dna carriers and solid-state nanopores
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4901370/
https://www.ncbi.nlm.nih.gov/pubmed/27121643
http://dx.doi.org/10.1021/acs.nanolett.6b00627
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