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顺磁性纳米颗粒作为基于荧光共振能量转移的肌氨酸皮摩尔检测平台。

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

作者信息

Heger Zbynek, Cernei Natalia, Krizkova Sona, Masarik Michal, Kopel Pavel, Hodek Petr, Zitka Ondrej, Adam Vojtech, Kizek Rene

机构信息

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.

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

出版信息

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

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.

摘要

在此,我们描述了一种基于Förster共振能量转移(FRET)的超灵敏特异性生物传感系统,用于检测肌氨酸作为前列腺癌的潜在生物标志物。该FRET生物传感器利用固定在顺磁性纳米颗粒表面的抗肌氨酸抗体进行特异性抗原结合。肌氨酸的成功结合导致由抗肌氨酸抗体组成的夹心结构的组装,使FRET对的Förster距离(Ro)保持在所需的接近度。检测基于金功能化绿色荧光蛋白与抗体@量子点生物共轭物之间的光谱重叠(激发波长400 nm)。基于FRET效率(F₆₀₄/F₅₁₀比值)的肌氨酸饱和曲线在5至50 nM的线性动态范围内进行了测试,检测限低至50 pM。然后,组装好的生物传感器成功用于前列腺癌细胞系(PC3、22Rv1、PNT1A)和前列腺腺癌患者尿液样本中肌氨酸的定量分析。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/6b4c/4352859/92145fb2f66a/srep08868-f1.jpg

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