scholarly journals An Enzyme- and Label-Free Fluorescence Aptasensor for Detection of Thrombin Based on Graphene Oxide and G-Quadruplex

Sensors ◽  
2019 ◽  
Vol 19 (20) ◽  
pp. 4424 ◽  
Author(s):  
Yani Wei ◽  
Luhui Wang ◽  
Yingying Zhang ◽  
Yafei Dong

An enzyme- and label-free aptamer-based assay is described for the determination of thrombin. A DNA strand (S) consisting of two parts was designed, where the first (Sa) is the thrombin-binding aptamer and the second (Se) is a G-quadruplex. In the absence of thrombin, Sa is readily adsorbed by graphene oxide (GO), which has a preference for ss-DNA rather than for ds-DNA. Upon the addition of the N-methyl-mesoporphyrin IX (NMM), its fluorescence (with excitation/emission at 399/610 nm) is quenched by GO. In contrast, in the presence of thrombin, the aptamer will bind thrombin, and thus, be separated from GO. As a result, fluorescence will be enhanced. The increase is linear in the 0.37 µM to 50 µM thrombin concentration range, and the detection limit is 0.37 nM. The method is highly selective over other proteins, cost-effective, and simple. In our perception, it represents a universal detection scheme that may be applied to other targets according to the proper choice of the aptamer sequence and formation of a suitable aptamer-target pair.


Biosensors ◽  
2021 ◽  
Vol 11 (3) ◽  
pp. 85
Author(s):  
Wassa Waiwinya ◽  
Thitirat Putnin ◽  
Dechnarong Pimalai ◽  
Wireeya Chawjiraphan ◽  
Nuankanya Sathirapongsasuti ◽  
...  

An immobilization-free electrochemical sensor coupled with a graphene oxide (GO)-based aptasensor was developed for glycated human serum albumin (GHSA) detection. The concentration of GHSA was monitored by measuring the electrochemical response of free GO and aptamer-bound GO in the presence of glycated albumin; their currents served as the analytical signals. The electrochemical aptasensor exhibited good performance with a base-10 logarithmic scale. The calibration curve was achieved in the range of 0.01–50 µg/mL. The limit of detection (LOD) was 8.70 ng/mL. The developed method was considered a one-drop measurement process because a fabrication step and the probe-immobilization process were not required. This simple sensor offers a cost-effective, rapid, and sensitive detection method, and could be an alternative approach for determination of GHSA levels.



RSC Advances ◽  
2021 ◽  
Vol 11 (20) ◽  
pp. 12227-12234
Author(s):  
Hisham S. M. Abd-Rabboh ◽  
Abd El-Galil E. Amr ◽  
Elsayed A. Elsayed ◽  
Ahmed Y. A. Sayed ◽  
Ayman H. Kamel

Robust, reliable and cost-effective paper-based analytical device for potentiometric pholcodine (opiate derivative drug) ion sensing has been prepared and characterized.



2014 ◽  
Vol 6 (20) ◽  
pp. 8120-8123 ◽  
Author(s):  
Yunfeng Bai ◽  
Lu Zhao ◽  
Zezhong Chen ◽  
Haiyan Wang ◽  
Feng Feng


2020 ◽  
Vol 187 (5) ◽  
Author(s):  
Jagriti Sethi ◽  
Michiel Van Bulck ◽  
Ahmed Suhail ◽  
Mina Safarzadeh ◽  
Ana Perez-Castillo ◽  
...  

AbstractA label-free biosensor is developed for the determination of plasma-based Aβ1–42 biomarker in Alzheimer’s disease (AD). The platform is based on highly conductive dual-layer of graphene and electrochemically reduced graphene oxide (rGO). The modification of dual-layer with 1-pyrenebutyric acid N-hydroxysuccinimide ester (Pyr-NHS) is achieved to facilitate immobilization of H31L21 antibody. The effect of these modifications were studied with morphological, spectral and electrochemical techniques. The response of the biosensor was evaluated using differential pulse voltammetry (DPV). The data was acquired at a working potential of ~ 180 mV and a scan rate of 50 mV s−1. A low limit of detection (LOD) of 2.398 pM is achieved over a wide linear range from 11 pM to 55 nM. The biosensor exhibits excellent specificity over Aβ1–40 and ApoE ε4 interfering species. Thus, it provides a viable tool for electrochemical determination of Aβ1–42. Spiked human and mice plasmas were used for the successful validation of the sensing platform in bio-fluidic samples. The results obtained from mice plasma analysis concurred with the immunohistochemistry (IHC) and magnetic resonance imaging (MRI) data obtained from brain analysis.



2015 ◽  
Vol 7 (22) ◽  
pp. 9615-9618 ◽  
Author(s):  
Yahui Guo ◽  
Yan Sun ◽  
Xiaoqiang Shen ◽  
Xing Chen ◽  
Weirong Yao ◽  
...  

A label-free fluorescence assay was developed for the detection of Zn2+ using GO and a guanine-rich sequence H22.



Molecules ◽  
2021 ◽  
Vol 26 (23) ◽  
pp. 7352
Author(s):  
Yun Zhang ◽  
Xinao Ma ◽  
Jingtian Zhang ◽  
Feixian Luo ◽  
Wenshu Wang ◽  
...  

G-quadruplexes can bind with hemin to form peroxidase-like DNAzymes that are widely used in the design of biosensors. However, the catalytic activity of G-quadruplex/hemin DNAzyme is relatively low compared with natural peroxidase, which hampers its sensitivity and, thus, its application in the detection of nucleic acids. In this study, we developed a high-sensitivity biosensor targeting norovirus nucleic acids through rationally introducing a dimeric G-quadruplex structure into the DNAzyme. In this strategy, two separate molecular beacons each having a G-quadruplex-forming sequence embedded in the stem structure are brought together through hybridization with a target DNA strand, and thus forms a three-way junction architecture and allows a dimeric G-quadruplex to form, which, upon binding with hemin, has a synergistic enhancement of catalytic activities. This provides a high-sensitivity colorimetric readout by the catalyzing H2O2-mediated oxidation of 2,2′-azino-bis(3-ethylbenzothiazoline -6-sulfonic acid) diammonium salt (ABTS). Up to 10 nM of target DNA can be detected through colorimetric observation with the naked eye using our strategy. Hence, our approach provides a non-amplifying, non-labeling, simple-operating, cost-effective colorimetric biosensing method for target nucleic acids, such as norovirus-conserved sequence detection, and highlights the further implication of higher-order multimerized G-quadruplex structures in the design of high-sensitivity biosensors.



Biosensors ◽  
2020 ◽  
Vol 10 (3) ◽  
pp. 24 ◽  
Author(s):  
Shanshan Wei ◽  
Haolin Xiao ◽  
Liangli Cao ◽  
Zhencheng Chen

In this article, a highly sensitive label-free immunosensor based on a graphene oxide (GO)/Fe3O4/Prussian blue (PB) nanocomposite modified electrode was developed for the determination of human hepatitis B surface antigen (HBsAg). In this electrochemical immunoassay system, PB was used as a redox probe, while GO/Fe3O4/PB nanocomposites and AuNPs were prepared and coated on screen-printed electrodes to enhance the detection sensitivity and to immobilize the hepatitis B surface antibody (HBsAb). The immunosensor was fabricated based on the principle that the decrease in peak currents of PB is proportional to the concentration of HBsAg captured on the modified immunosensor. The experimental results revealed that the immunosensor exhibited a sensitive response to HBsAg in the range of 0.5 pg·mL−1 to 200 ng·mL−1, and with a low detection limit of 0.166 pg·mL−1 (S/N = 3). Furthermore, the proposed immunosensor was used to detect several clinical serum samples with acceptable results, and it also showed good reproducibility, selectivity and stability, which may have a promising potential application in clinical immunoassays.



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