Simultaneous Detection of L‐Lactate and D‐Glucose Using DNA Aptamers in Human Blood Serum**

Huang, Po‐Jung Jimmy; Liu, Juewen

doi:10.1002/anie.202212879

Cited by 54 publications

(29 citation statements)

References 53 publications

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“…Recently, many new aptamers have been reported, [ 33 , 35 , 36 , 37 ] and the development of reliable binding assays is ever more important. [ 1 , 2 , 38 ] In this work, we examined the stability of the fluorescence signal of various DNA staining dyes with various aptamers in both polystyrene microplates and a quartz cuvette.…”

Section: Discussionmentioning

confidence: 99%

Surfactant-Assisted Label-Free Fluorescent Aptamer Biosensors and Binding Assays

Zhang

Liu

2023

Biosensors

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Using DNA staining dyes such as SYBR Green I (SGI) and thioflavin T (ThT) to perform label-free detection of aptamer binding has been performed for a long time for both binding assays and biosensor development. Since these dyes are cationic, they can also adsorb to the wall of reaction vessels leading to unstable signals and even false interpretations of the results. In this work, the stability of the signal was first evaluated using ThT and the classic adenosine aptamer. In a polystyrene microplate, a drop in fluorescence was observed even when non-binding targets or water were added, whereas a more stable signal was achieved in a quartz cuvette. Equilibrating the system can also improve signal stability. In addition, a few polymers and surfactants were also screened, and 0.01% Triton X-100 was found to have the best protection effect against fluorescence signal decrease due to dye adsorption. Three aptamers for Hg2+, adenosine, and cortisol were tested for their sensitivity and signal stability in the absence and presence of Triton X-100. In each case, the sensitivity was similar, whereas the signal stability was better for the surfactant. This study indicates that careful control experiments need to be designed to ensure reliable results and that the reliability can be improved by using Triton X-100 and a long equilibration time.

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Section: Discussionmentioning

confidence: 99%

Surfactant-Assisted Label-Free Fluorescent Aptamer Biosensors and Binding Assays

Zhang

Liu

2023

Biosensors

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“…In addition to artificial small molecules that act as carbohydrate-binding agents via non-covalent bonding, DNA and lectin have also been explored [ 63 ]. Glycosylation, a prevalent post-translational modification of proteins and lipids in eukaryotic cells, has been associated with neurological diseases and brain cancers due to abnormal cell surface glycan expression [ 64 ].…”

Section: Dna and Lectin-based Probesmentioning

confidence: 99%

Sensitive and specific detection of saccharide species based on fluorescence: update from 2016

Huang

Han

et al. 2023

Anal Bioanal Chem

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Increasing evidence supports the critical role of saccharides in various pathophysiological steps of tumor progression, where they regulate tumor proliferation, invasion, hematogenic metastasis, and angiogenesis. The identification and recognition of these saccharides provide a solid foundation for the development of targeted drug preparations, which are however not fully understood due to their complex and similar structures. In order to achieve fluorescence sensing of saccharides, extensive research has been conducted to design molecular probes and nanoparticles made of different materials. This paper aims to provide in-depth discussion of three main topics that cover the current status of the carbohydrate sensing based on the fluorescence sensing mechanism, including a phenylboronic acid-based sensing platform, non-boronic acid entities, as well as an enzyme-based sensing platform. It also highlights efforts made to understand the recognition mechanisms and improve the sensing properties of these systems. Finally, we present the challenge of achieving high selectivity and sensitivity recognition of saccharides, and suggest possible future avenues for exploration. Graphical Abstract

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“…By immobilizing a DNA library that can fold to a hairpin structure, − we recently used adenosine and ATP as targets and also observed this classical aptamer especially in the early rounds of the selections when the target concentrations were high . In the late rounds, when the target concentrations were dropped to as low as 1 μM, this sequence was suppressed and two higher affinity aptamers ( K d ∼ 200 nM adenosine) dominated.…”

Section: Introductionmentioning

confidence: 99%

Cross-Binding of Four Adenosine/ATP Aptamers to Caffeine, Theophylline, and Other Methylxanthines

Ding

Xie

et al. 2023

Biochemistry

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The classical DNA aptamer for adenosine and ATP was selected twice using ATP as the target in 1995 and 2005, respectively. In 2022, this motif appeared four more times from selections using adenosine, ATP, theophylline, and caffeine as targets, suggesting that this aptamer can also bind methylxanthines. In this work, using thioflavin T fluorescence spectroscopy, this classical DNA aptamer showed K d values for adenosine, theophylline, and caffeine of 9.5, 101, and 131 μM, respectively, and similar K d values were obtained using isothermal titration calorimetry. Binding to the methylxanthines was also observed for the newly selected Ade1301 aptamer but not for the Ade1304 aptamer. The RNA aptamer for ATP also had no binding to the methylxanthines. Molecular dynamics simulations were performed using the classical DNA and RNA aptamers based on their NMR structures, and the simulation results were consistent with the experimental observations, explaining the selectivity profiles. This study suggests that a broader range of target analogues need to be tested for aptamers. For the detection of adenosine and ATP, the Ade1304 aptamer is a better choice due to its better selectivity.

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Simultaneous Detection of L‐Lactate and D‐Glucose Using DNA Aptamers in Human Blood Serum**

Cited by 54 publications

References 53 publications

Surfactant-Assisted Label-Free Fluorescent Aptamer Biosensors and Binding Assays

Surfactant-Assisted Label-Free Fluorescent Aptamer Biosensors and Binding Assays

Sensitive and specific detection of saccharide species based on fluorescence: update from 2016

Cross-Binding of Four Adenosine/ATP Aptamers to Caffeine, Theophylline, and Other Methylxanthines

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