scholarly journals A Label-Free Fluorescent Assay for the Rapid and Sensitive Detection of Adenosine Deaminase Activity and Inhibition

Sensors ◽  
2018 ◽  
Vol 18 (8) ◽  
pp. 2441 ◽  
Author(s):  
Xinxing Tang ◽  
Kefeng Wu ◽  
Han Zhao ◽  
Mingjian Chen ◽  
Changbei Ma
Keyword(s):  
Adenosine Deaminase ◽  
High Selectivity ◽  
High Sensitivity ◽  
Rnase H ◽  
Fluorescence Signal ◽  
Label Free ◽  
Fluorescent Assay ◽  
Adenosine Deaminase Activity ◽  
G Quadruplex ◽  
Almost All

Adenosine deaminase (ADA), able to catalyze the irreversible deamination of adenosine into inosine, can be found in almost all tissues and plays an important role in several diseases. In this work, we developed a label-free fluorescence method for the detection of adenosine deaminase activity and inhibition. In the presence of ADA, ATP has been shown to be hydrolyzed. The ATP aptamer was shown to form a G-quadruplex/thioflavin T (ThT) complex with ThT and exhibited an obvious fluorescence signal. However, the ATP aptamer could bind with ATP and exhibited a low fluorescence signal because of the absence of ADA. This assay showed high sensitivity to ADA with a detection limit of 1 U/L based on an SNR of 3 and got a good linear relationship within the range of 1–100 U/L with R2 = 0.9909. The LOD is lower than ADA cutoff value (4 U/L) in the clinical requirement and more sensitive than most of the reported methods. This technique exhibited high selectivity for ADA against hoGG I, UDG, RNase H and λexo. Moreover, this strategy was successfully applied for assaying the inhibition of ADA using erythro-9-(2-hydroxy-3-nonyl) adenine (EHNA) and, as such, demonstrated great potential for the future use in the diagnosis of ADA-relevant diseases, particularly in advanced drug development.

A label-free and enzyme-free signal amplification strategy for a sensitive RNase H activity assay

Nanoscale ◽  
2017 ◽  
Vol 9 (42) ◽  
pp. 16149-16153 ◽  
Author(s):  
Chang Yeol Lee ◽  
Hyowon Jang ◽  
Ki Soo Park ◽  
Hyun Gyu Park
Keyword(s):  
Signal Amplification ◽  
Rnase H ◽  
Label Free ◽  
Activity Assay ◽  
G Quadruplex ◽  
Catalytic Hairpin Assembly ◽  
Amplification Strategy

A target-triggered catalytic hairpin assembly with a G-quadruplex specific fluorescent binder, NMM, is employed to develop a novel and sensitive RNase H activity assay.

A Label-Free Fluorescent AND Logic Gate Aptasensor for Carbohydrate Antigen 15-3 Detection Based on Graphene Oxide

2021 ◽  
Vol 24 ◽  
Author(s):  
Wenxiao Hu ◽  
Yafei Dong ◽  
Luhui Wang ◽  
Yue Wang ◽  
Mengyao Qian ◽  
...  
Keyword(s):  
Fluorescent Dyes ◽  
Logic Gate ◽  
High Sensitivity ◽  
Carbohydrate Antigen ◽  
Fluorescence Signal ◽  
Label Free ◽  
Molecular Logic ◽  
Long Stem ◽  
Middle Ring ◽  
Fluorescent Aptasensor

Background: Molecular logic gate always used fluorescent dyes to realize fluorescence signal. The labeling of the fluorophore is relatively expensive, low yield and singly labeled impuritiesaffects the affinity between the target and the aptamer. Label-free fluorescent aptamer biosensor strategy has attracted widespread interest due to lower cost and simple. Objective: Herein, we have designed a AND logic gate fluorescent aptasensor for detecting carbohydrate antigen 15-3(CA15-3) based on label-free fluorescence signal output. Materials and Methods: A hairpin DNA probe consists of CA15-3 aptamer and partly anti-CA15-3 aptamer sequences as a long stem and G-rich sequences of the middle ring as a quadruplex-forming oligomer. G-rich sequences can fold into a quadruplex by K+, and then G-quadruplex interacts specifically with N-methylmesoporphyrin IX(NMM), leading to a dramatic increase in fluorescence of NMM. With CA15-3 and NMM as the two inputs, the fluorescence intensity of the NMM is the output signal. Lacking of CA15-3 or NMM, there is no significant fluorescence enhancing, and the output of the signal is “0”. The fluorescence signal was dramatically increasing and the output of the signal is “1” only when CA15-3 protein and NMM were added at the same time. Results: This biosensor strategy possessed selectivity, high sensitivity for detecting CA15-3 protein from 10 to 500 U mL-1 and the detection limit was 10 U mL-1, and also showed good reproducibility in spiked human serum. Conclusion: In summary, the proposed AND logic gate fluorescent aptasensor could specifically detect CA15-3.

A “turn-on” and label-free fluorescent assay for the rapid detection of exonuclease III activity based on Tb3+-induced G-quadruplex conjugates

2014 ◽  
Vol 406 (18) ◽  
pp. 4535-4540 ◽  
Author(s):  
WeiJuan Yang ◽  
YaJuan Ruan ◽  
WeiHua Wu ◽  
PingPing Chen ◽  
LiangJun Xu ◽  
...  
Keyword(s):  
Rapid Detection ◽  
Label Free ◽  
Fluorescent Assay ◽  
Exonuclease Iii ◽  
Turn On ◽  
G Quadruplex

Improved Detection of Hg2+ in Aqueous Solution with High Sensitivity and Selectivity by Using Mercury-Specific DNA, Sybr Green I and Gold Nanoparticles

2011 ◽  
Vol 239-242 ◽  
pp. 934-939
Author(s):  
Hui Xu ◽  
Shuli Gao ◽  
Jian Nong Chen ◽  
Quan Wen Liu
Keyword(s):  
Gold Nanoparticles ◽  
High Sensitivity ◽  
Sybr Green ◽  
Sybr Green I ◽  
Fluorescence Signal ◽  
Potassium Ions ◽  
Label Free ◽  
Turn On ◽  
Sensitivity And Selectivity ◽  
Fluorescence Turn On

We report a label-free, fast, fluorescence turn on assay for Hg2+detecton by using mercury-specific DNA (MSD), Sybr Green I (SG) and gold nanoparticles (AuNPs). SG efficiently discriminates MSD and MSD/Hg2+complex. The addition of gold nanoparticle decreases the background fluorescence signal further for MSD. The fluorescence intensity of MSD/Hg2+complex keeps constant after addition of AuNPs. This property improves the signal-to-background ratio and decreases the detection limitation further. In addition, the method shows improved selectivity compared with that in the absence of AuNPs. This strategy could be applied to the detection of potassium ions and showed good generality.

Effect of adenosine deaminase on cardiac interstitial adenosine

1992 ◽  
Vol 263 (4) ◽  
pp. H1322-H1326 ◽  
Author(s):  
Q. Zhu ◽  
G. P. Matherne ◽  
R. R. Curnish ◽  
C. G. Tribble ◽  
R. M. Berne
Keyword(s):  
Guinea Pig ◽  
Adenosine Deaminase ◽  
Interstitial Fluid ◽  
Aortic Pressure ◽  
Guinea Pig Heart ◽  
Coronary Pressure ◽  
Flow Perfusion ◽  
Adenosine Deaminase Activity ◽  
Coronary Tone ◽  
Almost All

Adenosine deaminase was infused into isolated perfused guinea pig hearts to determine its effect on myocardial adenosine levels. The enzyme was administered during constant coronary flow perfusion at 6.11 +/- 0.36 ml.min-1.g-1. Venous adenosine was measured in samples of pulmonary artery effluent; epicardial and endocardial adenosine were measured with the porous nylon disk technique. Infusion of adenosine deaminase at 2.4 and 4.8 U/ml produced adenosine deaminase activity of 0.92 +/- 0.09 and 2.33 +/- 0.15 U/ml, respectively, in epicardial fluid and 1.93 +/- 0.28 and 4.84 +/- 0.47 U/ml, respectively, in endocardial fluid. Aortic pressure was unchanged by infusion of adenosine deaminase at both infusion rates. Adenosine deaminase (data from both infusion rates pooled) reduced epicardial adenosine from 0.327 +/- 0.028 to 0.139 +/- 0.022 microM, endocardial adenosine from 4.61 +/- 0.42 to 1.64 +/- 0.20 microM, and venous adenosine from 0.017 +/- 0.02 to 0.003 +/- 0.001 microM. The data indicate that infused adenosine deaminase reaches the epicardial and endocardial interstitial fluid (ISF) compartments. The absence of any effect on coronary pressure suggests that adenosine may not be involved in resting basal coronary tone. The presence of significant residual adenosine despite adenosine deaminase infusion indicates that adenosine production in the unstressed isolated guinea pig heart exceeds the degradative capacity of infused adenosine deaminase. Previous studies in which it was assumed that almost all of the endogenous adenosine is inactivated by the infusion of adenosine deaminase should be reevaluated in light of these observations.

A label-free ultrasensitive and selective strategy for Pb(ii) assay by a multifunctional DNA probe-mediated rolling-circle amplified synthesis of the G-quadruplexes

2018 ◽  
Vol 10 (25) ◽  
pp. 3081-3088 ◽  
Author(s):  
Xiao-Feng Wang ◽  
Yong-Sheng Wang ◽  
Xi-Lin Xiao ◽  
Wen-Bo Lan ◽  
Bin Zhou ◽  
...  
Keyword(s):  
Dna Probe ◽  
Fluorescence Signal ◽  
Label Free ◽  
Rolling Circle ◽  
G Quadruplex ◽  
Selective Strategy ◽  
Dna Template

The cleavage of the S-DNA in a MDP by Pb(ii) can release an E-DNA, which initiates a RCA reaction with a padlock DNA template. The formed G-quadruplex could specifically bind to NMM to result in an amplified fluorescence signal.

scholarly journals Layer-by-layer biofunctionalization of nanostructured porous silicon for high-sensitivity and high-selectivity label-free affinity biosensing

2018 ◽  
Vol 9 (1) ◽  
Author(s):  
Stefano Mariani ◽  
Valentina Robbiano ◽  
Lucanos M. Strambini ◽  
Aline Debrassi ◽  
Gabriela Egri ◽  
...  
Keyword(s):  
Porous Silicon ◽  
High Selectivity ◽  
High Sensitivity ◽  
Layer By Layer ◽  
Label Free

Turn-on and label-free fluorescence detection of lead ions based on target-induced G-quadruplex formation

2015 ◽  
Vol 51 (38) ◽  
pp. 8165-8168 ◽  
Author(s):  
Lijun Xu ◽  
Xiaoqiang Shen ◽  
Shanni Hong ◽  
Jine Wang ◽  
Yuanyuan Zhang ◽  
...  
Keyword(s):  
Fluorescence Detection ◽  
High Sensitivity ◽  
Lead Ions ◽  
Label Free ◽  
Low Background ◽  
Background Fluorescence ◽  
Turn On ◽  
Sensing Systems ◽  
G Quadruplex ◽  
Quadruplex Formation

Using a guanine-rich sequence (AGRO100) and N-methyl mesoporphyrin IX (NMM), a turn-on and label-free fluorescent Pb2+ sensor with high sensitivity and low background fluorescence was presented as a representative of five turn-on sensing systems.

A luminescent G-quadruplex-selective iridium(iii) complex for the label-free detection of lysozyme

2016 ◽  
Vol 4 (14) ◽  
pp. 2407-2411 ◽  
Author(s):  
Lihua Lu ◽  
Wanhe Wang ◽  
Modi Wang ◽  
Tian-Shu Kang ◽  
Jin-Jian Lu ◽  
...  
Keyword(s):  
High Selectivity ◽  
Label Free ◽  
Label Free Detection ◽  
G Quadruplex

A novel Ir(iii) complex 1 displays high selectivity for the G-quadruplex, and was used to establish a label-free G-quadruplex-based detection platform for lysozyme in buffer.

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