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

See more in PubMed

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

Cernei N. et al. Sarcosine as a Potential Prostate Cancer Biomarker-A Review. Int J Mol Sci 14, 13893–13908 (2013). PubMed PMC

Heger Z. et al. Determination of common urine substances as an assay for improving prostate carcinoma diagnostics. Oncol Rep 31, 1846–1854 (2014). PubMed

Sreekumar A. et al. Metabolomic profiles delineate potential role for sarcosine in prostate cancer progression. Nature 457, 910–914 (2009). 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. et al. Photoelectrochemical Sensor Based on Quantum Dots and Sarcosine Oxidase. ChemPhysChem 14, 2338–2342 (2013). 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. et al. Colorimetric determination of sarcosine in urine samples of prostatic carcinoma by mimic enzyme palladium nanoparticles. Anal Chim Acta 825, 63–68 (2014). 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. et al. Optimized Protocol of Chicken Antibody (IgY) Purification Providing Electrophoretically Homogenous Preparations. Int J Electrochem Sci 8, 113–124 (2013).

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. et al. Nanoparticle-Mediated Cytoplasmic Delivery of Proteins To Target Cellular Machinery. ACS Nano 4, 1493–1500 (2010). PubMed

Love J. C. et al. Self-assembled monolayers of thiolates on metals as a form of nanotechnology. Chem Rev 105, 1103–1169 (2005). 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. et al. Electrophoretic study of peptide-mediated quantum dot-human immunoglobulin bioconjugation. Electrophoresis 34, 2725–2732 (2013). PubMed

Casanova D. et al. Single lanthanide-doped oxide nanoparticles as donors in fluorescence resonance energy transfer experiments. J Phys Chem B 110, 19264–19270 (2006). PubMed

Khan A. P. et al. The Role of Sarcosine Metabolism in Prostate Cancer Progression. Neoplasia 15, 491–501 (2013). 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. et al. Microfluidic chip coupled with modified paramagnetic particles for sarcosine isolation in urine. Electrophoresis 34, 2639–2647 (2013). PubMed

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