Plasmonic AuNP/g‑C3N4 Nanohybrid-based Photoelectrochemical Sensing Platform for Ultrasensitive Monitoring of Polynucleotide Kinase Activity Accompanying DNAzyme-Catalyzed Precipitation Amplification
A convenient and feasible photoelectrochemical (PEC) sensing platform based on gold nanoparticles-decorated g-C3N4 nanosheets (AuNP/g-C3N4) was designed for highly sensitive monitoring of T4 polynucleotide kinase (PNK) activity, using DNAzyme-mediated catalytic precipitation amplification. To realiz...
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Published in | ACS applied materials & interfaces Vol. 7; no. 15; pp. 8330 - 8338 |
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Main Authors | , , , , |
Format | Journal Article |
Language | English |
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United States
American Chemical Society
22.04.2015
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Abstract | A convenient and feasible photoelectrochemical (PEC) sensing platform based on gold nanoparticles-decorated g-C3N4 nanosheets (AuNP/g-C3N4) was designed for highly sensitive monitoring of T4 polynucleotide kinase (PNK) activity, using DNAzyme-mediated catalytic precipitation amplification. To realize our design, the AuNP/g-C3N4 nanohybrid was initially synthesized through in situ reduction of Au(III) on the g-C3N4 nanosheets, which was utilized for the immobilization of hairpin DNA1 (HP1) on the sensing interface. Thereafter, a target-induced isothermal amplification was automatically carried out on hairpin DNA2 (HP2) in the solution phase through PNK-catalyzed 5′-phosphorylation accompanying formation of numerous trigger DNA fragments, which could induce generation of hemin/G-quadruplex-based DNAzyme on hairpin DNA1. Subsequently, the DNAzyme could catalyze the 4-chloro-1-naphthol (4-CN) oxidation to produce an insoluble precipitation on the AuNP/g-C3N4 surface, thereby resulting in the local alternation of the photocurrent. Experimental results revealed that introduction of AuNP on the g-C3N4 could cause a ∼100% increase in the photocurrent because of surface plasmon resonance-enhanced light harvesting and separation of photogenerated e–/h+ pairs. Under the optimal conditions, the percentage of photocurrent decrement (ΔI/I 0, relative to background signal) increased with the increasing PNK activity in a dynamic working range from 2 to 100 mU mL–1 with a low detection limit (LOD) of 1.0 mU mL–1. The inhibition effect of adenosine diphosphate also received a good performance in PNK inhibitor screening research, thereby providing a useful scheme for practical use in quantitative PNK activity assay for life science and biological research. |
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AbstractList | A convenient and feasible photoelectrochemical (PEC) sensing platform based on gold nanoparticles-decorated g-C3N4 nanosheets (AuNP/g-C3N4) was designed for highly sensitive monitoring of T4 polynucleotide kinase (PNK) activity, using DNAzyme-mediated catalytic precipitation amplification. To realize our design, the AuNP/g-C3N4 nanohybrid was initially synthesized through in situ reduction of Au(III) on the g-C3N4 nanosheets, which was utilized for the immobilization of hairpin DNA1 (HP1) on the sensing interface. Thereafter, a target-induced isothermal amplification was automatically carried out on hairpin DNA2 (HP2) in the solution phase through PNK-catalyzed 5′-phosphorylation accompanying formation of numerous trigger DNA fragments, which could induce generation of hemin/G-quadruplex-based DNAzyme on hairpin DNA1. Subsequently, the DNAzyme could catalyze the 4-chloro-1-naphthol (4-CN) oxidation to produce an insoluble precipitation on the AuNP/g-C3N4 surface, thereby resulting in the local alternation of the photocurrent. Experimental results revealed that introduction of AuNP on the g-C3N4 could cause a ∼100% increase in the photocurrent because of surface plasmon resonance-enhanced light harvesting and separation of photogenerated e–/h+ pairs. Under the optimal conditions, the percentage of photocurrent decrement (ΔI/I 0, relative to background signal) increased with the increasing PNK activity in a dynamic working range from 2 to 100 mU mL–1 with a low detection limit (LOD) of 1.0 mU mL–1. The inhibition effect of adenosine diphosphate also received a good performance in PNK inhibitor screening research, thereby providing a useful scheme for practical use in quantitative PNK activity assay for life science and biological research. A convenient and feasible photoelectrochemical (PEC) sensing platform based on gold nanoparticles-decorated g-C3N4 nanosheets (AuNP/g-C3N4) was designed for highly sensitive monitoring of T4 polynucleotide kinase (PNK) activity, using DNAzyme-mediated catalytic precipitation amplification. To realize our design, the AuNP/g-C3N4 nanohybrid was initially synthesized through in situ reduction of Au(III) on the g-C3N4 nanosheets, which was utilized for the immobilization of hairpin DNA1 (HP1) on the sensing interface. Thereafter, a target-induced isothermal amplification was automatically carried out on hairpin DNA2 (HP2) in the solution phase through PNK-catalyzed 5'-phosphorylation accompanying formation of numerous trigger DNA fragments, which could induce generation of hemin/G-quadruplex-based DNAzyme on hairpin DNA1. Subsequently, the DNAzyme could catalyze the 4-chloro-1-naphthol (4-CN) oxidation to produce an insoluble precipitation on the AuNP/g-C3N4 surface, thereby resulting in the local alternation of the photocurrent. Experimental results revealed that introduction of AuNP on the g-C3N4 could cause a ∼100% increase in the photocurrent because of surface plasmon resonance-enhanced light harvesting and separation of photogenerated e-/h+ pairs. Under the optimal conditions, the percentage of photocurrent decrement (ΔI/I0, relative to background signal) increased with the increasing PNK activity in a dynamic working range from 2 to 100 mU mL(-1) with a low detection limit (LOD) of 1.0 mU mL(-1). The inhibition effect of adenosine diphosphate also received a good performance in PNK inhibitor screening research, thereby providing a useful scheme for practical use in quantitative PNK activity assay for life science and biological research. |
Author | Lai, Wenqiang Xu, Mingdi Zhou, Qian Tang, Dianping Zhuang, Junyang |
AuthorAffiliation | Department of Chemistry Fuzhou University |
AuthorAffiliation_xml | – name: Department of Chemistry – name: Fuzhou University |
Author_xml | – sequence: 1 givenname: Junyang surname: Zhuang fullname: Zhuang, Junyang – sequence: 2 givenname: Wenqiang surname: Lai fullname: Lai, Wenqiang – sequence: 3 givenname: Mingdi surname: Xu fullname: Xu, Mingdi – sequence: 4 givenname: Qian surname: Zhou fullname: Zhou, Qian – sequence: 5 givenname: Dianping surname: Tang fullname: Tang, Dianping email: dianping.tang@fzu.edu.cn |
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Keywords | gold-decorated g-C3N4 nanosheets photoelectrochemistry surface plasmon resonance T4 polynucleotide kinase isothermal amplification |
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SubjectTerms | Biosensing Techniques - instrumentation Catalysis Conductometry - instrumentation DNA, Catalytic - chemistry Enzyme Activation Equipment Design Equipment Failure Analysis Gold - chemistry Materials Testing Metal Nanoparticles - chemistry Metal Nanoparticles - ultrastructure Nitriles - chemistry Nucleic Acid Amplification Techniques - instrumentation Photochemistry - instrumentation Polynucleotide 5'-Hydroxyl-Kinase - analysis Polynucleotide 5'-Hydroxyl-Kinase - chemistry Surface Plasmon Resonance - instrumentation |
Title | Plasmonic AuNP/g‑C3N4 Nanohybrid-based Photoelectrochemical Sensing Platform for Ultrasensitive Monitoring of Polynucleotide Kinase Activity Accompanying DNAzyme-Catalyzed Precipitation Amplification |
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