Potentiometric sensing of nuclease activities and oxidative damage of single-stranded DNA using a polycation-sensitive membrane electrode
A simple, general and label-free potentiometric method to measure nuclease activities and oxidative DNA damage in a homogeneous solution using a polycation-sensitive membrane electrode is reported. Protamine, a linear polyionic species, is used as an indicator to report the cleavage of DNA by nuclea...
Ausführliche Beschreibung
Gespeichert in:
| Autor*in: |
Ding, Jiawang [verfasserIn] |
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| Format: |
E-Artikel |
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| Sprache: |
Englisch |
| Erschienen: |
2013transfer abstract |
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| Schlagwörter: |
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| Umfang: |
7 |
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| Übergeordnetes Werk: |
Enthalten in: Vertical differentiation via multi-tier geographical indications and the consumer perception of quality: The case of Chianti wines - Costanigro, Marco ELSEVIER, 2019, the principal international journal devoted to research, design development and application of biosensors and bioelectronics, Amsterdam [u.a.] |
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| Übergeordnetes Werk: |
volume:47 ; year:2013 ; day:15 ; month:09 ; pages:559-565 ; extent:7 |
| Links: |
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| DOI / URN: |
10.1016/j.bios.2013.03.066 |
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ELV022060898 |
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| 245 | 1 | 0 | |a Potentiometric sensing of nuclease activities and oxidative damage of single-stranded DNA using a polycation-sensitive membrane electrode |
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| 520 | |a A simple, general and label-free potentiometric method to measure nuclease activities and oxidative DNA damage in a homogeneous solution using a polycation-sensitive membrane electrode is reported. Protamine, a linear polyionic species, is used as an indicator to report the cleavage of DNA by nucleases such as restriction and nonspecific nucleases, and the damage of DNA induced by hydroxyl radicals. Measurements can be done with a titration mode or a direct detection mode. For the potentiometric titration mode, the enzymatic cleavage dramatically affects the electrostatical interaction between DNA and protamine and thus shifts the response curve for the potentiometric titration of the DNA with protamine. Under the optimized conditions, the enzyme activities can be sensed potentiometrically with detection limits of 2.7×10−4 U/µL for S1 nuclease, and of 3.9×10−4 U/µL for DNase I. For the direct detection mode, a biocomplex between protamine and DNA is used as a substrate. The nuclease of interest cleaves the DNA from the protamine/DNA complex into smaller fragments, so that free protamine is generated and can be detected potentiometrically via the polycation-sensitive membrane electrode. Using a direct measurement, the nuclease activities could be rapidly detected with detection limits of 3.2×10−4 U/µL for S1 nuclease, and of 4.5×10−4 U/µL for DNase I. Moreover, the proposed potentiometric assays demonstrate the potential applications in the detection of hydroxyl radicals. It is anticipated that the present potentiometric strategy will provide a promising platform for high-throughput screening of nucleases, reactive oxygen species and the drugs with potential inhibition abilities. | ||
| 520 | |a A simple, general and label-free potentiometric method to measure nuclease activities and oxidative DNA damage in a homogeneous solution using a polycation-sensitive membrane electrode is reported. Protamine, a linear polyionic species, is used as an indicator to report the cleavage of DNA by nucleases such as restriction and nonspecific nucleases, and the damage of DNA induced by hydroxyl radicals. Measurements can be done with a titration mode or a direct detection mode. For the potentiometric titration mode, the enzymatic cleavage dramatically affects the electrostatical interaction between DNA and protamine and thus shifts the response curve for the potentiometric titration of the DNA with protamine. Under the optimized conditions, the enzyme activities can be sensed potentiometrically with detection limits of 2.7×10−4 U/µL for S1 nuclease, and of 3.9×10−4 U/µL for DNase I. For the direct detection mode, a biocomplex between protamine and DNA is used as a substrate. The nuclease of interest cleaves the DNA from the protamine/DNA complex into smaller fragments, so that free protamine is generated and can be detected potentiometrically via the polycation-sensitive membrane electrode. Using a direct measurement, the nuclease activities could be rapidly detected with detection limits of 3.2×10−4 U/µL for S1 nuclease, and of 4.5×10−4 U/µL for DNase I. Moreover, the proposed potentiometric assays demonstrate the potential applications in the detection of hydroxyl radicals. It is anticipated that the present potentiometric strategy will provide a promising platform for high-throughput screening of nucleases, reactive oxygen species and the drugs with potential inhibition abilities. | ||
| 650 | 7 | |a Nucleases |2 Elsevier | |
| 650 | 7 | |a Polyion sensors |2 Elsevier | |
| 650 | 7 | |a Protamine |2 Elsevier | |
| 650 | 7 | |a Hydroxyl radicals |2 Elsevier | |
| 650 | 7 | |a Potentiometry |2 Elsevier | |
| 700 | 1 | |a Qin, Wei |4 oth | |
| 773 | 0 | 8 | |i Enthalten in |n Elsevier Science |a Costanigro, Marco ELSEVIER |t Vertical differentiation via multi-tier geographical indications and the consumer perception of quality: The case of Chianti wines |d 2019 |d the principal international journal devoted to research, design development and application of biosensors and bioelectronics |g Amsterdam [u.a.] |w (DE-627)ELV001931067 |
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10.1016/j.bios.2013.03.066 doi GBVA2013015000020.pica (DE-627)ELV022060898 (ELSEVIER)S0956-5663(13)00236-4 DE-627 ger DE-627 rakwb eng 570 610 570 DE-600 610 DE-600 630 640 VZ 49.00 bkl Ding, Jiawang verfasserin aut Potentiometric sensing of nuclease activities and oxidative damage of single-stranded DNA using a polycation-sensitive membrane electrode 2013transfer abstract 7 nicht spezifiziert zzz rdacontent nicht spezifiziert z rdamedia nicht spezifiziert zu rdacarrier A simple, general and label-free potentiometric method to measure nuclease activities and oxidative DNA damage in a homogeneous solution using a polycation-sensitive membrane electrode is reported. Protamine, a linear polyionic species, is used as an indicator to report the cleavage of DNA by nucleases such as restriction and nonspecific nucleases, and the damage of DNA induced by hydroxyl radicals. Measurements can be done with a titration mode or a direct detection mode. For the potentiometric titration mode, the enzymatic cleavage dramatically affects the electrostatical interaction between DNA and protamine and thus shifts the response curve for the potentiometric titration of the DNA with protamine. Under the optimized conditions, the enzyme activities can be sensed potentiometrically with detection limits of 2.7×10−4 U/µL for S1 nuclease, and of 3.9×10−4 U/µL for DNase I. For the direct detection mode, a biocomplex between protamine and DNA is used as a substrate. The nuclease of interest cleaves the DNA from the protamine/DNA complex into smaller fragments, so that free protamine is generated and can be detected potentiometrically via the polycation-sensitive membrane electrode. Using a direct measurement, the nuclease activities could be rapidly detected with detection limits of 3.2×10−4 U/µL for S1 nuclease, and of 4.5×10−4 U/µL for DNase I. Moreover, the proposed potentiometric assays demonstrate the potential applications in the detection of hydroxyl radicals. It is anticipated that the present potentiometric strategy will provide a promising platform for high-throughput screening of nucleases, reactive oxygen species and the drugs with potential inhibition abilities. A simple, general and label-free potentiometric method to measure nuclease activities and oxidative DNA damage in a homogeneous solution using a polycation-sensitive membrane electrode is reported. Protamine, a linear polyionic species, is used as an indicator to report the cleavage of DNA by nucleases such as restriction and nonspecific nucleases, and the damage of DNA induced by hydroxyl radicals. Measurements can be done with a titration mode or a direct detection mode. For the potentiometric titration mode, the enzymatic cleavage dramatically affects the electrostatical interaction between DNA and protamine and thus shifts the response curve for the potentiometric titration of the DNA with protamine. Under the optimized conditions, the enzyme activities can be sensed potentiometrically with detection limits of 2.7×10−4 U/µL for S1 nuclease, and of 3.9×10−4 U/µL for DNase I. For the direct detection mode, a biocomplex between protamine and DNA is used as a substrate. The nuclease of interest cleaves the DNA from the protamine/DNA complex into smaller fragments, so that free protamine is generated and can be detected potentiometrically via the polycation-sensitive membrane electrode. Using a direct measurement, the nuclease activities could be rapidly detected with detection limits of 3.2×10−4 U/µL for S1 nuclease, and of 4.5×10−4 U/µL for DNase I. Moreover, the proposed potentiometric assays demonstrate the potential applications in the detection of hydroxyl radicals. It is anticipated that the present potentiometric strategy will provide a promising platform for high-throughput screening of nucleases, reactive oxygen species and the drugs with potential inhibition abilities. Nucleases Elsevier Polyion sensors Elsevier Protamine Elsevier Hydroxyl radicals Elsevier Potentiometry Elsevier Qin, Wei oth Enthalten in Elsevier Science Costanigro, Marco ELSEVIER Vertical differentiation via multi-tier geographical indications and the consumer perception of quality: The case of Chianti wines 2019 the principal international journal devoted to research, design development and application of biosensors and bioelectronics Amsterdam [u.a.] (DE-627)ELV001931067 volume:47 year:2013 day:15 month:09 pages:559-565 extent:7 https://doi.org/10.1016/j.bios.2013.03.066 Volltext GBV_USEFLAG_U GBV_ELV SYSFLAG_U 49.00 Hauswirtschaft: Allgemeines VZ AR 47 2013 15 0915 559-565 7 045F 570 |
| spelling |
10.1016/j.bios.2013.03.066 doi GBVA2013015000020.pica (DE-627)ELV022060898 (ELSEVIER)S0956-5663(13)00236-4 DE-627 ger DE-627 rakwb eng 570 610 570 DE-600 610 DE-600 630 640 VZ 49.00 bkl Ding, Jiawang verfasserin aut Potentiometric sensing of nuclease activities and oxidative damage of single-stranded DNA using a polycation-sensitive membrane electrode 2013transfer abstract 7 nicht spezifiziert zzz rdacontent nicht spezifiziert z rdamedia nicht spezifiziert zu rdacarrier A simple, general and label-free potentiometric method to measure nuclease activities and oxidative DNA damage in a homogeneous solution using a polycation-sensitive membrane electrode is reported. Protamine, a linear polyionic species, is used as an indicator to report the cleavage of DNA by nucleases such as restriction and nonspecific nucleases, and the damage of DNA induced by hydroxyl radicals. Measurements can be done with a titration mode or a direct detection mode. For the potentiometric titration mode, the enzymatic cleavage dramatically affects the electrostatical interaction between DNA and protamine and thus shifts the response curve for the potentiometric titration of the DNA with protamine. Under the optimized conditions, the enzyme activities can be sensed potentiometrically with detection limits of 2.7×10−4 U/µL for S1 nuclease, and of 3.9×10−4 U/µL for DNase I. For the direct detection mode, a biocomplex between protamine and DNA is used as a substrate. The nuclease of interest cleaves the DNA from the protamine/DNA complex into smaller fragments, so that free protamine is generated and can be detected potentiometrically via the polycation-sensitive membrane electrode. Using a direct measurement, the nuclease activities could be rapidly detected with detection limits of 3.2×10−4 U/µL for S1 nuclease, and of 4.5×10−4 U/µL for DNase I. Moreover, the proposed potentiometric assays demonstrate the potential applications in the detection of hydroxyl radicals. It is anticipated that the present potentiometric strategy will provide a promising platform for high-throughput screening of nucleases, reactive oxygen species and the drugs with potential inhibition abilities. A simple, general and label-free potentiometric method to measure nuclease activities and oxidative DNA damage in a homogeneous solution using a polycation-sensitive membrane electrode is reported. Protamine, a linear polyionic species, is used as an indicator to report the cleavage of DNA by nucleases such as restriction and nonspecific nucleases, and the damage of DNA induced by hydroxyl radicals. Measurements can be done with a titration mode or a direct detection mode. For the potentiometric titration mode, the enzymatic cleavage dramatically affects the electrostatical interaction between DNA and protamine and thus shifts the response curve for the potentiometric titration of the DNA with protamine. Under the optimized conditions, the enzyme activities can be sensed potentiometrically with detection limits of 2.7×10−4 U/µL for S1 nuclease, and of 3.9×10−4 U/µL for DNase I. For the direct detection mode, a biocomplex between protamine and DNA is used as a substrate. The nuclease of interest cleaves the DNA from the protamine/DNA complex into smaller fragments, so that free protamine is generated and can be detected potentiometrically via the polycation-sensitive membrane electrode. Using a direct measurement, the nuclease activities could be rapidly detected with detection limits of 3.2×10−4 U/µL for S1 nuclease, and of 4.5×10−4 U/µL for DNase I. Moreover, the proposed potentiometric assays demonstrate the potential applications in the detection of hydroxyl radicals. It is anticipated that the present potentiometric strategy will provide a promising platform for high-throughput screening of nucleases, reactive oxygen species and the drugs with potential inhibition abilities. Nucleases Elsevier Polyion sensors Elsevier Protamine Elsevier Hydroxyl radicals Elsevier Potentiometry Elsevier Qin, Wei oth Enthalten in Elsevier Science Costanigro, Marco ELSEVIER Vertical differentiation via multi-tier geographical indications and the consumer perception of quality: The case of Chianti wines 2019 the principal international journal devoted to research, design development and application of biosensors and bioelectronics Amsterdam [u.a.] (DE-627)ELV001931067 volume:47 year:2013 day:15 month:09 pages:559-565 extent:7 https://doi.org/10.1016/j.bios.2013.03.066 Volltext GBV_USEFLAG_U GBV_ELV SYSFLAG_U 49.00 Hauswirtschaft: Allgemeines VZ AR 47 2013 15 0915 559-565 7 045F 570 |
| allfields_unstemmed |
10.1016/j.bios.2013.03.066 doi GBVA2013015000020.pica (DE-627)ELV022060898 (ELSEVIER)S0956-5663(13)00236-4 DE-627 ger DE-627 rakwb eng 570 610 570 DE-600 610 DE-600 630 640 VZ 49.00 bkl Ding, Jiawang verfasserin aut Potentiometric sensing of nuclease activities and oxidative damage of single-stranded DNA using a polycation-sensitive membrane electrode 2013transfer abstract 7 nicht spezifiziert zzz rdacontent nicht spezifiziert z rdamedia nicht spezifiziert zu rdacarrier A simple, general and label-free potentiometric method to measure nuclease activities and oxidative DNA damage in a homogeneous solution using a polycation-sensitive membrane electrode is reported. Protamine, a linear polyionic species, is used as an indicator to report the cleavage of DNA by nucleases such as restriction and nonspecific nucleases, and the damage of DNA induced by hydroxyl radicals. Measurements can be done with a titration mode or a direct detection mode. For the potentiometric titration mode, the enzymatic cleavage dramatically affects the electrostatical interaction between DNA and protamine and thus shifts the response curve for the potentiometric titration of the DNA with protamine. Under the optimized conditions, the enzyme activities can be sensed potentiometrically with detection limits of 2.7×10−4 U/µL for S1 nuclease, and of 3.9×10−4 U/µL for DNase I. For the direct detection mode, a biocomplex between protamine and DNA is used as a substrate. The nuclease of interest cleaves the DNA from the protamine/DNA complex into smaller fragments, so that free protamine is generated and can be detected potentiometrically via the polycation-sensitive membrane electrode. Using a direct measurement, the nuclease activities could be rapidly detected with detection limits of 3.2×10−4 U/µL for S1 nuclease, and of 4.5×10−4 U/µL for DNase I. Moreover, the proposed potentiometric assays demonstrate the potential applications in the detection of hydroxyl radicals. It is anticipated that the present potentiometric strategy will provide a promising platform for high-throughput screening of nucleases, reactive oxygen species and the drugs with potential inhibition abilities. A simple, general and label-free potentiometric method to measure nuclease activities and oxidative DNA damage in a homogeneous solution using a polycation-sensitive membrane electrode is reported. Protamine, a linear polyionic species, is used as an indicator to report the cleavage of DNA by nucleases such as restriction and nonspecific nucleases, and the damage of DNA induced by hydroxyl radicals. Measurements can be done with a titration mode or a direct detection mode. For the potentiometric titration mode, the enzymatic cleavage dramatically affects the electrostatical interaction between DNA and protamine and thus shifts the response curve for the potentiometric titration of the DNA with protamine. Under the optimized conditions, the enzyme activities can be sensed potentiometrically with detection limits of 2.7×10−4 U/µL for S1 nuclease, and of 3.9×10−4 U/µL for DNase I. For the direct detection mode, a biocomplex between protamine and DNA is used as a substrate. The nuclease of interest cleaves the DNA from the protamine/DNA complex into smaller fragments, so that free protamine is generated and can be detected potentiometrically via the polycation-sensitive membrane electrode. Using a direct measurement, the nuclease activities could be rapidly detected with detection limits of 3.2×10−4 U/µL for S1 nuclease, and of 4.5×10−4 U/µL for DNase I. Moreover, the proposed potentiometric assays demonstrate the potential applications in the detection of hydroxyl radicals. It is anticipated that the present potentiometric strategy will provide a promising platform for high-throughput screening of nucleases, reactive oxygen species and the drugs with potential inhibition abilities. Nucleases Elsevier Polyion sensors Elsevier Protamine Elsevier Hydroxyl radicals Elsevier Potentiometry Elsevier Qin, Wei oth Enthalten in Elsevier Science Costanigro, Marco ELSEVIER Vertical differentiation via multi-tier geographical indications and the consumer perception of quality: The case of Chianti wines 2019 the principal international journal devoted to research, design development and application of biosensors and bioelectronics Amsterdam [u.a.] (DE-627)ELV001931067 volume:47 year:2013 day:15 month:09 pages:559-565 extent:7 https://doi.org/10.1016/j.bios.2013.03.066 Volltext GBV_USEFLAG_U GBV_ELV SYSFLAG_U 49.00 Hauswirtschaft: Allgemeines VZ AR 47 2013 15 0915 559-565 7 045F 570 |
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10.1016/j.bios.2013.03.066 doi GBVA2013015000020.pica (DE-627)ELV022060898 (ELSEVIER)S0956-5663(13)00236-4 DE-627 ger DE-627 rakwb eng 570 610 570 DE-600 610 DE-600 630 640 VZ 49.00 bkl Ding, Jiawang verfasserin aut Potentiometric sensing of nuclease activities and oxidative damage of single-stranded DNA using a polycation-sensitive membrane electrode 2013transfer abstract 7 nicht spezifiziert zzz rdacontent nicht spezifiziert z rdamedia nicht spezifiziert zu rdacarrier A simple, general and label-free potentiometric method to measure nuclease activities and oxidative DNA damage in a homogeneous solution using a polycation-sensitive membrane electrode is reported. Protamine, a linear polyionic species, is used as an indicator to report the cleavage of DNA by nucleases such as restriction and nonspecific nucleases, and the damage of DNA induced by hydroxyl radicals. Measurements can be done with a titration mode or a direct detection mode. For the potentiometric titration mode, the enzymatic cleavage dramatically affects the electrostatical interaction between DNA and protamine and thus shifts the response curve for the potentiometric titration of the DNA with protamine. Under the optimized conditions, the enzyme activities can be sensed potentiometrically with detection limits of 2.7×10−4 U/µL for S1 nuclease, and of 3.9×10−4 U/µL for DNase I. For the direct detection mode, a biocomplex between protamine and DNA is used as a substrate. The nuclease of interest cleaves the DNA from the protamine/DNA complex into smaller fragments, so that free protamine is generated and can be detected potentiometrically via the polycation-sensitive membrane electrode. Using a direct measurement, the nuclease activities could be rapidly detected with detection limits of 3.2×10−4 U/µL for S1 nuclease, and of 4.5×10−4 U/µL for DNase I. Moreover, the proposed potentiometric assays demonstrate the potential applications in the detection of hydroxyl radicals. It is anticipated that the present potentiometric strategy will provide a promising platform for high-throughput screening of nucleases, reactive oxygen species and the drugs with potential inhibition abilities. A simple, general and label-free potentiometric method to measure nuclease activities and oxidative DNA damage in a homogeneous solution using a polycation-sensitive membrane electrode is reported. Protamine, a linear polyionic species, is used as an indicator to report the cleavage of DNA by nucleases such as restriction and nonspecific nucleases, and the damage of DNA induced by hydroxyl radicals. Measurements can be done with a titration mode or a direct detection mode. For the potentiometric titration mode, the enzymatic cleavage dramatically affects the electrostatical interaction between DNA and protamine and thus shifts the response curve for the potentiometric titration of the DNA with protamine. Under the optimized conditions, the enzyme activities can be sensed potentiometrically with detection limits of 2.7×10−4 U/µL for S1 nuclease, and of 3.9×10−4 U/µL for DNase I. For the direct detection mode, a biocomplex between protamine and DNA is used as a substrate. The nuclease of interest cleaves the DNA from the protamine/DNA complex into smaller fragments, so that free protamine is generated and can be detected potentiometrically via the polycation-sensitive membrane electrode. Using a direct measurement, the nuclease activities could be rapidly detected with detection limits of 3.2×10−4 U/µL for S1 nuclease, and of 4.5×10−4 U/µL for DNase I. Moreover, the proposed potentiometric assays demonstrate the potential applications in the detection of hydroxyl radicals. It is anticipated that the present potentiometric strategy will provide a promising platform for high-throughput screening of nucleases, reactive oxygen species and the drugs with potential inhibition abilities. Nucleases Elsevier Polyion sensors Elsevier Protamine Elsevier Hydroxyl radicals Elsevier Potentiometry Elsevier Qin, Wei oth Enthalten in Elsevier Science Costanigro, Marco ELSEVIER Vertical differentiation via multi-tier geographical indications and the consumer perception of quality: The case of Chianti wines 2019 the principal international journal devoted to research, design development and application of biosensors and bioelectronics Amsterdam [u.a.] (DE-627)ELV001931067 volume:47 year:2013 day:15 month:09 pages:559-565 extent:7 https://doi.org/10.1016/j.bios.2013.03.066 Volltext GBV_USEFLAG_U GBV_ELV SYSFLAG_U 49.00 Hauswirtschaft: Allgemeines VZ AR 47 2013 15 0915 559-565 7 045F 570 |
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10.1016/j.bios.2013.03.066 doi GBVA2013015000020.pica (DE-627)ELV022060898 (ELSEVIER)S0956-5663(13)00236-4 DE-627 ger DE-627 rakwb eng 570 610 570 DE-600 610 DE-600 630 640 VZ 49.00 bkl Ding, Jiawang verfasserin aut Potentiometric sensing of nuclease activities and oxidative damage of single-stranded DNA using a polycation-sensitive membrane electrode 2013transfer abstract 7 nicht spezifiziert zzz rdacontent nicht spezifiziert z rdamedia nicht spezifiziert zu rdacarrier A simple, general and label-free potentiometric method to measure nuclease activities and oxidative DNA damage in a homogeneous solution using a polycation-sensitive membrane electrode is reported. Protamine, a linear polyionic species, is used as an indicator to report the cleavage of DNA by nucleases such as restriction and nonspecific nucleases, and the damage of DNA induced by hydroxyl radicals. Measurements can be done with a titration mode or a direct detection mode. For the potentiometric titration mode, the enzymatic cleavage dramatically affects the electrostatical interaction between DNA and protamine and thus shifts the response curve for the potentiometric titration of the DNA with protamine. Under the optimized conditions, the enzyme activities can be sensed potentiometrically with detection limits of 2.7×10−4 U/µL for S1 nuclease, and of 3.9×10−4 U/µL for DNase I. For the direct detection mode, a biocomplex between protamine and DNA is used as a substrate. The nuclease of interest cleaves the DNA from the protamine/DNA complex into smaller fragments, so that free protamine is generated and can be detected potentiometrically via the polycation-sensitive membrane electrode. Using a direct measurement, the nuclease activities could be rapidly detected with detection limits of 3.2×10−4 U/µL for S1 nuclease, and of 4.5×10−4 U/µL for DNase I. Moreover, the proposed potentiometric assays demonstrate the potential applications in the detection of hydroxyl radicals. It is anticipated that the present potentiometric strategy will provide a promising platform for high-throughput screening of nucleases, reactive oxygen species and the drugs with potential inhibition abilities. A simple, general and label-free potentiometric method to measure nuclease activities and oxidative DNA damage in a homogeneous solution using a polycation-sensitive membrane electrode is reported. Protamine, a linear polyionic species, is used as an indicator to report the cleavage of DNA by nucleases such as restriction and nonspecific nucleases, and the damage of DNA induced by hydroxyl radicals. Measurements can be done with a titration mode or a direct detection mode. For the potentiometric titration mode, the enzymatic cleavage dramatically affects the electrostatical interaction between DNA and protamine and thus shifts the response curve for the potentiometric titration of the DNA with protamine. Under the optimized conditions, the enzyme activities can be sensed potentiometrically with detection limits of 2.7×10−4 U/µL for S1 nuclease, and of 3.9×10−4 U/µL for DNase I. For the direct detection mode, a biocomplex between protamine and DNA is used as a substrate. The nuclease of interest cleaves the DNA from the protamine/DNA complex into smaller fragments, so that free protamine is generated and can be detected potentiometrically via the polycation-sensitive membrane electrode. Using a direct measurement, the nuclease activities could be rapidly detected with detection limits of 3.2×10−4 U/µL for S1 nuclease, and of 4.5×10−4 U/µL for DNase I. Moreover, the proposed potentiometric assays demonstrate the potential applications in the detection of hydroxyl radicals. It is anticipated that the present potentiometric strategy will provide a promising platform for high-throughput screening of nucleases, reactive oxygen species and the drugs with potential inhibition abilities. Nucleases Elsevier Polyion sensors Elsevier Protamine Elsevier Hydroxyl radicals Elsevier Potentiometry Elsevier Qin, Wei oth Enthalten in Elsevier Science Costanigro, Marco ELSEVIER Vertical differentiation via multi-tier geographical indications and the consumer perception of quality: The case of Chianti wines 2019 the principal international journal devoted to research, design development and application of biosensors and bioelectronics Amsterdam [u.a.] (DE-627)ELV001931067 volume:47 year:2013 day:15 month:09 pages:559-565 extent:7 https://doi.org/10.1016/j.bios.2013.03.066 Volltext GBV_USEFLAG_U GBV_ELV SYSFLAG_U 49.00 Hauswirtschaft: Allgemeines VZ AR 47 2013 15 0915 559-565 7 045F 570 |
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potentiometric sensing of nuclease activities and oxidative damage of single-stranded dna using a polycation-sensitive membrane electrode |
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Potentiometric sensing of nuclease activities and oxidative damage of single-stranded DNA using a polycation-sensitive membrane electrode |
| abstract |
A simple, general and label-free potentiometric method to measure nuclease activities and oxidative DNA damage in a homogeneous solution using a polycation-sensitive membrane electrode is reported. Protamine, a linear polyionic species, is used as an indicator to report the cleavage of DNA by nucleases such as restriction and nonspecific nucleases, and the damage of DNA induced by hydroxyl radicals. Measurements can be done with a titration mode or a direct detection mode. For the potentiometric titration mode, the enzymatic cleavage dramatically affects the electrostatical interaction between DNA and protamine and thus shifts the response curve for the potentiometric titration of the DNA with protamine. Under the optimized conditions, the enzyme activities can be sensed potentiometrically with detection limits of 2.7×10−4 U/µL for S1 nuclease, and of 3.9×10−4 U/µL for DNase I. For the direct detection mode, a biocomplex between protamine and DNA is used as a substrate. The nuclease of interest cleaves the DNA from the protamine/DNA complex into smaller fragments, so that free protamine is generated and can be detected potentiometrically via the polycation-sensitive membrane electrode. Using a direct measurement, the nuclease activities could be rapidly detected with detection limits of 3.2×10−4 U/µL for S1 nuclease, and of 4.5×10−4 U/µL for DNase I. Moreover, the proposed potentiometric assays demonstrate the potential applications in the detection of hydroxyl radicals. It is anticipated that the present potentiometric strategy will provide a promising platform for high-throughput screening of nucleases, reactive oxygen species and the drugs with potential inhibition abilities. |
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A simple, general and label-free potentiometric method to measure nuclease activities and oxidative DNA damage in a homogeneous solution using a polycation-sensitive membrane electrode is reported. Protamine, a linear polyionic species, is used as an indicator to report the cleavage of DNA by nucleases such as restriction and nonspecific nucleases, and the damage of DNA induced by hydroxyl radicals. Measurements can be done with a titration mode or a direct detection mode. For the potentiometric titration mode, the enzymatic cleavage dramatically affects the electrostatical interaction between DNA and protamine and thus shifts the response curve for the potentiometric titration of the DNA with protamine. Under the optimized conditions, the enzyme activities can be sensed potentiometrically with detection limits of 2.7×10−4 U/µL for S1 nuclease, and of 3.9×10−4 U/µL for DNase I. For the direct detection mode, a biocomplex between protamine and DNA is used as a substrate. The nuclease of interest cleaves the DNA from the protamine/DNA complex into smaller fragments, so that free protamine is generated and can be detected potentiometrically via the polycation-sensitive membrane electrode. Using a direct measurement, the nuclease activities could be rapidly detected with detection limits of 3.2×10−4 U/µL for S1 nuclease, and of 4.5×10−4 U/µL for DNase I. Moreover, the proposed potentiometric assays demonstrate the potential applications in the detection of hydroxyl radicals. It is anticipated that the present potentiometric strategy will provide a promising platform for high-throughput screening of nucleases, reactive oxygen species and the drugs with potential inhibition abilities. |
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A simple, general and label-free potentiometric method to measure nuclease activities and oxidative DNA damage in a homogeneous solution using a polycation-sensitive membrane electrode is reported. Protamine, a linear polyionic species, is used as an indicator to report the cleavage of DNA by nucleases such as restriction and nonspecific nucleases, and the damage of DNA induced by hydroxyl radicals. Measurements can be done with a titration mode or a direct detection mode. For the potentiometric titration mode, the enzymatic cleavage dramatically affects the electrostatical interaction between DNA and protamine and thus shifts the response curve for the potentiometric titration of the DNA with protamine. Under the optimized conditions, the enzyme activities can be sensed potentiometrically with detection limits of 2.7×10−4 U/µL for S1 nuclease, and of 3.9×10−4 U/µL for DNase I. For the direct detection mode, a biocomplex between protamine and DNA is used as a substrate. The nuclease of interest cleaves the DNA from the protamine/DNA complex into smaller fragments, so that free protamine is generated and can be detected potentiometrically via the polycation-sensitive membrane electrode. Using a direct measurement, the nuclease activities could be rapidly detected with detection limits of 3.2×10−4 U/µL for S1 nuclease, and of 4.5×10−4 U/µL for DNase I. Moreover, the proposed potentiometric assays demonstrate the potential applications in the detection of hydroxyl radicals. It is anticipated that the present potentiometric strategy will provide a promising platform for high-throughput screening of nucleases, reactive oxygen species and the drugs with potential inhibition abilities. |
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