Development of a sensitive enzyme-linked immunosorbent assay for measurement of DNase I in human serum

Nakajima, Tamiko; Takagi, Ryukichi; Tsutsumi, Yutaka; Makita, Chikako; Kominato, Yoshihiko; Kuribara, Jun; Ohshima, Shigeru; Tada, Hiroshi; Tsurugaya, Hideki; Kobayashi, Yasuyuki; Takeshita, Hitoshi; Kawai, Yasuyuki; Yasuda, Toshihiro

doi:10.1016/j.cca.2009.03.005

Cited by 19 publications

(4 citation statements)

References 17 publications

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“…The endeavors to increase the sensitivity of measurement continued, and Nakajima et al developed the sensitive enzyme-linked immunosorbent assay (ELISA) for the measurement of DNase I in human serum. This sandwich type of ELISA measured the DNase I protein using a polyclonal antibody against this protein and a biotinylated monoclonal antibody for the detection [ 90 ]. However, it is clear that ELISA is used to quantify the protein, in this case the enzyme, but not its activity, which may be dependent on activators and inhibitors and not only on the quantity of the enzyme.…”

Section: Methods Of the Measurement Of Dnase Activitymentioning

confidence: 99%

Deoxyribonucleases and Their Applications in Biomedicine

et al. 2020

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Extracellular DNA, also called cell-free DNA, released from dying cells or activated immune cells can be recognized by the immune system as a danger signal causing or enhancing inflammation. The cleavage of extracellular DNA is crucial for limiting the inflammatory response and maintaining homeostasis. Deoxyribonucleases (DNases) as enzymes that degrade DNA are hypothesized to play a key role in this process as a determinant of the variable concentration of extracellular DNA. DNases are divided into two families—DNase I and DNase II, according to their biochemical and biological properties as well as the tissue-specific production. Studies have shown that low DNase activity is both, a biomarker and a pathogenic factor in systemic lupus erythematosus. Interventional experiments proved that administration of exogenous DNase has beneficial effects in inflammatory diseases. Recombinant human DNase reduces mucus viscosity in lungs and is used for the treatment of patients with cystic fibrosis. This review summarizes the currently available published data about DNases, their activity as a potential biomarker and methods used for their assessment. An overview of the experiments with systemic administration of DNase is also included. Whether low-plasma DNase activity is involved in the etiopathogenesis of diseases remains unknown and needs to be elucidated.

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Section: Methods Of the Measurement Of Dnase Activitymentioning

confidence: 99%

Deoxyribonucleases and Their Applications in Biomedicine

et al. 2020

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“…Recently, novel approaches such as microelectrophoresis [19] and enzyme-linked immunosorbent assay (ELISA)-based assays [20,21] have also been reported.…”

Section: Introductionmentioning

confidence: 99%

Determination of DNase activity by degradation of ethidium bromide–DNA complexes using a fluorescence plate reader

Vogel

Frantz

2015

Analytical Biochemistry

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“…Traditional methods for the determination of DNase I activity include single radial enzyme diffusion (SRED) method, 9 electrochemical detection, 10 enzyme-linked immunosorbent assay (ELISA), 11 microchip electrophoresis 12 and uorescent probes. 13 Among them, the SRED method is convenient and highly sensitive, but requires approximately 20 h to complete a single test.…”

Section: Introductionmentioning

confidence: 99%

A label-free near-infrared fluorescent assay for the determination of deoxyribonuclease I activity based on malachite green/G-quadruplexes

Sun

Wang

Yan

2013

Analyst

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Owing to the biological and clinical significance of deoxyribonuclease I (DNase I), it is highly desirable to develop near-infrared (NIR) fluorescent assays for the determination of DNase I activity. Here we report a label-free NIR fluorescent assay for selective determination of DNase I activity based on malachite green (MG)/G-quadruplexes. In the presence of Na(+) or K(+), single stranded DNA (ssDNA) is able to form a G-quadruplex structure, thus to increase the rigidity of MG structure and result in a remarkable NIR fluorescence. As DNase I is capable of cleaving all types of DNA indiscriminately to release nucleotide products, the G-quadruplexes are cleaved into oligonucleotides in the presence of DNase I. As a result, the rigidity of MG structure is reduced, and the NIR fluorescence of the solution decreases with increase of DNase I activity, providing a useful platform for low-cost, label-free and convenient detection of DNase I activity. Under the optimum conditions, the proposed label-free NIR fluorescent assay gave a detection limit of 1 u mL(-1), and a relative standard deviation of 3.2% for eleven replicate detections of 50 u mL(-1) DNase I. The proposed assay was applied to the determination of DNase I activity in spiked human urine samples with recoveries from 99.1 to 109.0%.

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Development of a sensitive enzyme-linked immunosorbent assay for measurement of DNase I in human serum

Cited by 19 publications

References 17 publications

Deoxyribonucleases and Their Applications in Biomedicine

Deoxyribonucleases and Their Applications in Biomedicine

Determination of DNase activity by degradation of ethidium bromide–DNA complexes using a fluorescence plate reader

A label-free near-infrared fluorescent assay for the determination of deoxyribonuclease I activity based on malachite green/G-quadruplexes

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