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Sci Rep. 2015 Mar 9;5:8868. doi: 10.1038/srep08868.

Paramagnetic nanoparticles as a platform for FRET-based sarcosine picomolar detection.

Author information

1
1] Department of Chemistry and Biochemistry, Faculty of Agronomy, Mendel University in Brno, Zemedelska 1, CZ-613 00, Czech Republic, European Union [2] Central European Institute of Technology, Brno University of Technology, Technicka 3058/10, CZ-616 00 Brno, Czech Republic, European Union.
2
Department of Pathological Physiology, Faculty of Medicine, Masaryk University, Kamenice 5, CZ-612 00 Brno, Czech Republic, European Union.
3
Department of Biochemistry, Faculty of Science, Charles University in Prague, Hlavova 2030, CZ-128 40 Prague 2, Czech Republic, European Union.

Abstract

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 (λex 400 nm). The saturation curve of sarcosine based on FRET efficiency (F₆₀₄/F₅₁₀ 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.

PMID:
25746688
PMCID:
PMC4352859
DOI:
10.1038/srep08868
[Indexed for MEDLINE]
Free PMC Article

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