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Porous nanozymes: the peroxidase-mimetic activity of mesoporous iron oxide for the colorimetric and electrochemical detection of global DNA methylation†
Journal of Materials Chemistry B ( IF 6.1 ) Pub Date : 2018-06-22 00:00:00 , DOI: 10.1039/c8tb01132j Ripon Bhattacharjee 1, 2, 3, 4, 5 , Shunsuke Tanaka 6, 7, 8, 9, 10 , Sofia Moriam 1, 2, 3, 4, 5 , Mostafa Kamal Masud 1, 2, 3, 4, 5 , Jianjian Lin 6, 7, 8, 9 , Saad M. Alshehri 11, 12, 13, 14, 15 , Tansir Ahamad 11, 12, 13, 14, 15 , Rahul R. Salunkhe 16, 17, 18, 19 , Nam-Trung Nguyen 1, 2, 3, 4, 5 , Yusuke Yamauchi 5, 20, 21, 22, 23 , Md. Shahriar A. Hossain 5, 20, 21, 22, 23 , Muhammad J. A. Shiddiky 1, 2, 3, 4, 5
Journal of Materials Chemistry B ( IF 6.1 ) Pub Date : 2018-06-22 00:00:00 , DOI: 10.1039/c8tb01132j Ripon Bhattacharjee 1, 2, 3, 4, 5 , Shunsuke Tanaka 6, 7, 8, 9, 10 , Sofia Moriam 1, 2, 3, 4, 5 , Mostafa Kamal Masud 1, 2, 3, 4, 5 , Jianjian Lin 6, 7, 8, 9 , Saad M. Alshehri 11, 12, 13, 14, 15 , Tansir Ahamad 11, 12, 13, 14, 15 , Rahul R. Salunkhe 16, 17, 18, 19 , Nam-Trung Nguyen 1, 2, 3, 4, 5 , Yusuke Yamauchi 5, 20, 21, 22, 23 , Md. Shahriar A. Hossain 5, 20, 21, 22, 23 , Muhammad J. A. Shiddiky 1, 2, 3, 4, 5
Affiliation
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Nanomaterials (nanozymes) with peroxidase-mimetic activity have been widely used in biosensing platforms as low-cost, relatively stable and prevailing alternatives to natural enzymes. Herein, we report on the synthesis and application of the peroxidase-mimetic activity of mesoporous iron oxide (MIO) for the detection of global DNA methylation in colorectal cancer cell lines. The target DNA was extracted and denatured to get ssDNA followed by direct adsorption onto the surface of a bare screen-printed gold electrode (SPGE). A 5-methylcytosine antibody (5mC) functionalized nanomaterial (MIO-5mC) was then used to recognise the methylcytosine groups present on the SPGE. The MIO-5mC conjugates catalyse the TMB solution in the presence of hydrogen peroxide to give the colorimetric (i.e., naked-eye observation) and electrochemical detection of DNA methylation. The assay could successfully detect as low as 10% difference in the global DNA methylation level in synthetic samples and cell lines with good reproducibility and specificity (%RSD = <5%, for n = 3). This strategy avoids the use of natural enzyme horseradish peroxidase (HRP), traditional PCR based amplification and bisulfite treatment steps that are generally used in many conventional DNA methylation assays. We envisage that our assay could be a low-cost platform with great potential for genome-wide DNA methylation analysis in point-of-care applications.
中文翻译：
多孔纳米酶：介孔氧化铁的过氧化物酶模拟活性，用于比色和电化学检测整体DNA甲基化†
具有过氧化物酶模拟活性的纳米材料（纳米酶）已被广泛用作生物传感平台，是低成本，相对稳定且占优势的天然酶替代品。在此，我们报道了介孔氧化铁（MIO）的过氧化物酶模拟活性的合成和应用，用于检测结直肠癌细胞系中的整体DNA甲基化。提取目标DNA并变性，以获得ssDNA，然后直接吸附到裸丝网印刷金电极（SPGE）的表面上。然后使用5-甲基胞嘧啶抗体（5mC）功能化的纳米材料（MIO-5mC）识别SPGE上存在的甲基胞嘧啶基团。MIO-5mC共轭物在过氧化氢的存在下催化TMB溶液，以提供比色法（即，肉眼观察）和DNA甲基化的电化学检测。该方法可以成功地检测出合成样品和细胞系中总体DNA甲基化水平低至10％的差异，并且具有良好的可重复性和特异性（％RSD = <5％，n = 3）。该策略避免使用天然酶辣根过氧化物酶（HRP），传统的基于PCR的扩增和亚硫酸氢盐处理步骤，这些步骤通常在许多常规DNA甲基化测定中使用。我们设想，我们的检测方法可能是一种低成本平台，在即时医疗应用中对全基因组DNA甲基化分析具有巨大潜力。
更新日期：2018-06-22
中文翻译：
多孔纳米酶：介孔氧化铁的过氧化物酶模拟活性，用于比色和电化学检测整体DNA甲基化†
具有过氧化物酶模拟活性的纳米材料（纳米酶）已被广泛用作生物传感平台，是低成本，相对稳定且占优势的天然酶替代品。在此，我们报道了介孔氧化铁（MIO）的过氧化物酶模拟活性的合成和应用，用于检测结直肠癌细胞系中的整体DNA甲基化。提取目标DNA并变性，以获得ssDNA，然后直接吸附到裸丝网印刷金电极（SPGE）的表面上。然后使用5-甲基胞嘧啶抗体（5mC）功能化的纳米材料（MIO-5mC）识别SPGE上存在的甲基胞嘧啶基团。MIO-5mC共轭物在过氧化氢的存在下催化TMB溶液，以提供比色法（即，肉眼观察）和DNA甲基化的电化学检测。该方法可以成功地检测出合成样品和细胞系中总体DNA甲基化水平低至10％的差异，并且具有良好的可重复性和特异性（％RSD = <5％，n = 3）。该策略避免使用天然酶辣根过氧化物酶（HRP），传统的基于PCR的扩增和亚硫酸氢盐处理步骤，这些步骤通常在许多常规DNA甲基化测定中使用。我们设想，我们的检测方法可能是一种低成本平台，在即时医疗应用中对全基因组DNA甲基化分析具有巨大潜力。
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