Cargando…

Paramagnetic Nanoparticles as a Platform for FRET-Based Sarcosine Picomolar Detection

Herein, we describe an ultrasensitive specific biosensing system for detection of sarcosine as a potential biomarker of prostate carcinoma based on Förster resonance energy transfer (FRET). The FRET biosensor employs anti-sarcosine antibodies immobilized on paramagnetic nanoparticles surface for spe...

Descripción completa

Detalles Bibliográficos
Autores principales: Heger, Zbynek, Cernei, Natalia, Krizkova, Sona, Masarik, Michal, Kopel, Pavel, Hodek, Petr, Zitka, Ondrej, Adam, Vojtech, Kizek, Rene
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Nature Publishing Group 2015
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4352859/
https://www.ncbi.nlm.nih.gov/pubmed/25746688
http://dx.doi.org/10.1038/srep08868
_version_ 1782360514423160832
author Heger, Zbynek
Cernei, Natalia
Krizkova, Sona
Masarik, Michal
Kopel, Pavel
Hodek, Petr
Zitka, Ondrej
Adam, Vojtech
Kizek, Rene
author_facet Heger, Zbynek
Cernei, Natalia
Krizkova, Sona
Masarik, Michal
Kopel, Pavel
Hodek, Petr
Zitka, Ondrej
Adam, Vojtech
Kizek, Rene
author_sort Heger, Zbynek
collection PubMed
description Herein, we describe an ultrasensitive specific biosensing system for detection of sarcosine as a potential biomarker of prostate carcinoma based on Förster resonance energy transfer (FRET). The FRET biosensor employs anti-sarcosine antibodies immobilized on paramagnetic nanoparticles surface for specific antigen binding. Successful binding of sarcosine leads to assembly of a sandwich construct composed of anti-sarcosine antibodies keeping the Förster distance (Ro) of FRET pair in required proximity. The detection is based on spectral overlap between gold-functionalized green fluorescent protein and antibodies@quantum dots bioconjugate (λ(ex) 400 nm). The saturation curve of sarcosine based on FRET efficiency (F(604)/F(510) ratio) was tested within linear dynamic range from 5 to 50 nM with detection limit down to 50 pM. Assembled biosensor was then successfully employed for sarcosine quantification in prostatic cell lines (PC3, 22Rv1, PNT1A), and urinary samples of prostate adenocarcinoma patients.
format Online
Article
Text
id pubmed-4352859
institution National Center for Biotechnology Information
language English
publishDate 2015
publisher Nature Publishing Group
record_format MEDLINE/PubMed
spelling pubmed-43528592015-03-17 Paramagnetic Nanoparticles as a Platform for FRET-Based Sarcosine Picomolar Detection Heger, Zbynek Cernei, Natalia Krizkova, Sona Masarik, Michal Kopel, Pavel Hodek, Petr Zitka, Ondrej Adam, Vojtech Kizek, Rene Sci Rep Article Herein, we describe an ultrasensitive specific biosensing system for detection of sarcosine as a potential biomarker of prostate carcinoma based on Förster resonance energy transfer (FRET). The FRET biosensor employs anti-sarcosine antibodies immobilized on paramagnetic nanoparticles surface for specific antigen binding. Successful binding of sarcosine leads to assembly of a sandwich construct composed of anti-sarcosine antibodies keeping the Förster distance (Ro) of FRET pair in required proximity. The detection is based on spectral overlap between gold-functionalized green fluorescent protein and antibodies@quantum dots bioconjugate (λ(ex) 400 nm). The saturation curve of sarcosine based on FRET efficiency (F(604)/F(510) ratio) was tested within linear dynamic range from 5 to 50 nM with detection limit down to 50 pM. Assembled biosensor was then successfully employed for sarcosine quantification in prostatic cell lines (PC3, 22Rv1, PNT1A), and urinary samples of prostate adenocarcinoma patients. Nature Publishing Group 2015-03-09 /pmc/articles/PMC4352859/ /pubmed/25746688 http://dx.doi.org/10.1038/srep08868 Text en Copyright © 2015, Macmillan Publishers Limited. All rights reserved http://creativecommons.org/licenses/by/4.0/ This work is licensed under a Creative Commons Attribution 4.0 International License. The images or other third party material in this article are included in the article's Creative Commons license, unless indicated otherwise in the credit line; if the material is not included under the Creative Commons license, users will need to obtain permission from the license holder in order to reproduce the material. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/
spellingShingle Article
Heger, Zbynek
Cernei, Natalia
Krizkova, Sona
Masarik, Michal
Kopel, Pavel
Hodek, Petr
Zitka, Ondrej
Adam, Vojtech
Kizek, Rene
Paramagnetic Nanoparticles as a Platform for FRET-Based Sarcosine Picomolar Detection
title Paramagnetic Nanoparticles as a Platform for FRET-Based Sarcosine Picomolar Detection
title_full Paramagnetic Nanoparticles as a Platform for FRET-Based Sarcosine Picomolar Detection
title_fullStr Paramagnetic Nanoparticles as a Platform for FRET-Based Sarcosine Picomolar Detection
title_full_unstemmed Paramagnetic Nanoparticles as a Platform for FRET-Based Sarcosine Picomolar Detection
title_short Paramagnetic Nanoparticles as a Platform for FRET-Based Sarcosine Picomolar Detection
title_sort paramagnetic nanoparticles as a platform for fret-based sarcosine picomolar detection
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4352859/
https://www.ncbi.nlm.nih.gov/pubmed/25746688
http://dx.doi.org/10.1038/srep08868
work_keys_str_mv AT hegerzbynek paramagneticnanoparticlesasaplatformforfretbasedsarcosinepicomolardetection
AT cerneinatalia paramagneticnanoparticlesasaplatformforfretbasedsarcosinepicomolardetection
AT krizkovasona paramagneticnanoparticlesasaplatformforfretbasedsarcosinepicomolardetection
AT masarikmichal paramagneticnanoparticlesasaplatformforfretbasedsarcosinepicomolardetection
AT kopelpavel paramagneticnanoparticlesasaplatformforfretbasedsarcosinepicomolardetection
AT hodekpetr paramagneticnanoparticlesasaplatformforfretbasedsarcosinepicomolardetection
AT zitkaondrej paramagneticnanoparticlesasaplatformforfretbasedsarcosinepicomolardetection
AT adamvojtech paramagneticnanoparticlesasaplatformforfretbasedsarcosinepicomolardetection
AT kizekrene paramagneticnanoparticlesasaplatformforfretbasedsarcosinepicomolardetection