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.

] 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

Department of Biochemistry Faculty of Science Charles University Prague Hlavova 2030 CZ 128 40 Prague 2 Czech Republic European Union

Department of Pathological Physiology Faculty of Medicine Masaryk University Kamenice 5 CZ 612 00 Brno Czech Republic European Union

Zobrazit více v PubMed

Siegel R., Naishadham D. & Jemal A. Cancer statistics, 2013. CA-Cancer J Clin 63, 11–30 (2013). PubMed

Cernei N. PubMed PMC

Heger Z. PubMed

Sreekumar A. PubMed PMC

Jiang Y. Q., Cheng X. L., Wang C. A. & Ma Y. F. Quantitative Determination of Sarcosine and Related Compounds in Urinary Samples by Liquid Chromatography with Tandem Mass Spectrometry. Anal Chem 82, 9022–9027 (2010). PubMed

Riedel M. PubMed

Burton C., Gamagedara S. & Ma Y. F. A novel enzymatic technique for determination of sarcosine in urine samples. Anal Methods 4, 141–146 (2012).

Lan J. M. PubMed

Chen J., Zhang J., Zhang W. P. & Chen Z. L. Sensitive determination of the potential biomarker sarcosine for prostate cancer by LC-MS with N,N '-dicyclohexylcarbodiimide derivatization. J Sep Sci 37, 14–19 (2014). PubMed

Haviv A. H., Greneche J. M. & Lellouche J. P. Aggregation Control of Hydrophilic Maghemite (gamma-Fe2O3) Nanoparticles by Surface Doping Using Cerium Atoms. J Am Chem Soc 132, 12519–12521 (2010). PubMed

Guo Q. J., Teng X. W., Rahman S. & Yang H. Patterned Langmuir-Blodgett films of mondisperse nanoparticles of iron oxide using soft lithography. J Am Chem Soc 125, 630–631 (2003). PubMed

El-Boubbou K., Gruden C. & Huang X. Magnetic glyco-nanoparticles: A unique tool for rapid pathogen detection, decontamination, and strain differentiation. J Am Chem Soc 129, 13392–13393 (2007). PubMed

Schwegmann H., Feitz A. J. & Frimmel F. H. Influence of the zeta potential on the sorption and toxicity of iron oxide nanoparticles on S. cerevisiae and E. coli. J Colloid Interface Sci 347, 43–48 (2010). PubMed

Hodek P.

Shaner N. C., Steinbach P. A. & Tsien R. Y. A guide to choosing fluorescent proteins. Nat Methods 2, 905–909 (2005). PubMed

Bale S. S. PubMed

Love J. C. PubMed

Qu L. H. & Peng X. G. Control of photoluminescence properties of CdSe nanocrystals in growth. J Am Chem Soc 124, 2049–2055 (2002). PubMed

Janu L. PubMed

Casanova D. PubMed

Khan A. P. PubMed PMC

Ryan D., Robards K., Prenzler P. D. & Kendall M. Recent and potential developments in the analysis of urine: A review. Anal Chim Acta 684, 17–29 (2011). PubMed

Abate-Shen C. & Shen M. M. Diagnostics The prostate-cancer metabolome. Nature 457, 799–800 (2009). PubMed

Zitka O. PubMed

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