Dataset Information

Thrombin Determination Using Graphene Oxide Sensors with Co-Assisted Amplification.

ABSTRACT: Graphene oxide (GO) is widely used in sensors. The detection of proteins based on bare GO has been developed; however, the detection sensitivity needs to be improved. In this paper, a novel GO-DNA sensor for thrombin detection was developed using an aptamer linked to the surface of GO. Polyethylene glycol (PEG) was further used to prevent thrombin from nonspecific adsorption and to improve the sensitivity of the sensor for detection of thrombin. In order to improve the limit of detection for thrombin, we developed a GO and RecJf exonuclease co-assisted signal amplification strategy, and a detection limit of 24.35 fM for thrombin was achieved using this strategy. The results show that it is a promising method in analytical applications.

SUBMITTER: Liu L

PROVIDER: S-EPMC9502102 | biostudies-literature | 2022 Aug

REPOSITORIES: biostudies-literature

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Thrombin Determination Using Graphene Oxide Sensors with Co-Assisted Amplification.

Liu Lei L Li Qin Q Shi Haixia H Gao Li L

Micromachines 20220831 9

Graphene oxide (GO) is widely used in sensors. The detection of proteins based on bare GO has been developed; however, the detection sensitivity needs to be improved. In this paper, a novel GO-DNA sensor for thrombin detection was developed using an aptamer linked to the surface of GO. Polyethylene glycol (PEG) was further used to prevent thrombin from nonspecific adsorption and to improve the sensitivity of the sensor for detection of thrombin. In order to improve the limit of detection for thr ...[more]

PMID: 36144058

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Project description:Graphene oxide (GO), a two-dimensional material with a high aspect ratio and polar functional groups, can physically adsorb single-strand DNA through different types of interactions, such as hydrogen bonding and π-π stacking, making it an attractive nanocarrier for nucleic acids. In this work, we demonstrate a strategy to target exosites I and II of thrombin simultaneously by using programmed hybrid-aptamers for enhanced anticoagulation efficiency and stability. The targeting ligand is denoted as Supra-TBA15/29 (supramolecular TBA15/29), containing TBA15 (a 15-base nucleotide, targeting exosite I of thrombin) and TBA29 (a 29-base nucleotide, targeting exosite II of thrombin), and it is designed to allow consecutive hybridization of TBA15 and TBA29 to form a network of TBAs (i.e., supra-TBA15/29). The programmed hybrid-aptamers (Supra-TBA15/29) were self-assembled on GO to further boost anticoagulation activity by inhibiting thrombin activity, and thus suppress the thrombin-induced fibrin formation from fibrinogen. The Supra-TBA15/29-GO composite was formed mainly through multivalent interaction between poly(adenine) from Supra-TBA15/29 and GO. We controlled the assembly of Supra-TBA15/29 on GO by regulating the preparation temperature and the concentration ratio of Supra-TBA15/29 to GO to optimize the distance between TBA15 and TBA29 units, aptamer density, and aptamer orientation on the GO surfaces. The dose-dependent thrombin clotting time (TCT) delay caused by Supra-TBA15/29-GO was >10 times longer than that of common anticoagulant drugs including heparin, argatroban, hirudin, and warfarin. Supra-TBA15/29-GO exhibits high biocompatibility, which has been proved by in vitro cytotoxicity and hemolysis assays. In addition, the thromboelastography of whole-blood coagulation and rat-tail bleeding assays indicate the anticoagulation ability of Supra-TBA15/29-GO is superior to the most widely used anticoagulant (heparin). Our highly biocompatible Supra-TBA15/29-GO with strong multivalent interaction with thrombin [dissociation constant (K d) = 1.9 × 10-11 M] shows great potential as an effective direct thrombin inhibitor for the treatment of hemostatic disorders.

Dataset Information

Thrombin Determination Using Graphene Oxide Sensors with Co-Assisted Amplification.

Publications

Thrombin Determination Using Graphene Oxide Sensors with Co-Assisted Amplification.

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