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Target-induced cyclic DNAzyme formation for colorimetric and chemiluminescence imaging assay of protein biomarkers
Analyst ( IF 3.6 ) Pub Date : 2017-08-16 00:00:00 , DOI: 10.1039/c7an00413c Kaili Yang 1, 2, 3, 4, 5 , Min Huo 1, 2, 3, 4, 5 , Yuehua Guo 1, 2, 3, 4, 5 , Yizhuo Yang 1, 2, 3, 4, 5 , Jie Wu 1, 2, 3, 4, 5 , Lin Ding 1, 2, 3, 4, 5 , Huangxian Ju 1, 2, 3, 4, 5
Analyst ( IF 3.6 ) Pub Date : 2017-08-16 00:00:00 , DOI: 10.1039/c7an00413c Kaili Yang 1, 2, 3, 4, 5 , Min Huo 1, 2, 3, 4, 5 , Yuehua Guo 1, 2, 3, 4, 5 , Yizhuo Yang 1, 2, 3, 4, 5 , Jie Wu 1, 2, 3, 4, 5 , Lin Ding 1, 2, 3, 4, 5 , Huangxian Ju 1, 2, 3, 4, 5
Affiliation
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A target-induced cyclic strategy for DNAzyme formation was proposed to achieve simple, sensitive and universal detection of protein biomarkers with convenient colorimetric or chemiluminescence imaging readout. In the assay, the target protein was recognized by a pair of DNA-labeled antibodies (Ab1-DNA1 and Ab2-DNA2) to form a proximate complex, which could hybridize with the conjugate DNA3/DNA4 to release the guanine-rich DNA4 and thus formed G-quadruplex/hemin horseradish peroxidase-mimicking DNAzyme. The process could be further recycled with Exonuclease III by cleaving DNA3 to free the proximate complex, resulting in the cyclic formation of DNAzyme. The G-quadruplex/hemin DNAzyme could catalyze the H2O2-mediated oxidation of 3,3,5,5-tetramethylbenzidine to produce the color change from colorless to blue or enhance the chemiluminescence of a luminol–H2O2 system. Thus the signal could be read out with the naked eye, and by colorimetry and chemiluminescence imaging. Using a carcinoembryonic antigen as a model target, the proposed assay showed a detection range of 4 orders of magnitude along with detection limits of 170 and 16 pg mL−1 for colorimetric and chemiluminescence imaging assays respectively. This assay had the advantages of easy operation, sensitive detection, target flexibility and diversified signal readout, providing a great opportunity for commercial application.
中文翻译：
目标诱导的环状DNA酶的形成，用于比色法和化学发光成像法检测蛋白质生物标志物
提出了一种目标诱导的DNAzyme形成的循环策略，以简便的比色法或化学发光成像读数实现蛋白质生物标志物的简单，灵敏和通用检测。在测定中，靶蛋白被一对DNA标记的抗体（Ab1-DNA1和Ab2-DNA2）识别以形成邻近的复合物，该复合物可与缀合物DNA3 / DNA4杂交以释放富含鸟嘌呤的DNA4，从而形成了G-四链体/血红素辣根过氧化物酶模拟DNAzyme。通过切割DNA3释放邻近的复合物，可以用核酸外切酶III进一步循环利用该过程，从而导致DNA酶的循环形成。G-quadruplex / hemin DNAzyme可以催化H 2 O 2介导的3,3,5,5-四甲基联苯胺的氧化产生从无色到蓝色的颜色变化或增强luminol–H 2 O 2系统的化学发光。因此，可以用裸眼，比色法和化学发光成像读出信号。以癌胚抗原为模型靶标，所提出的测定法显示了比色法和化学发光成像测定法的检测范围为4个数量级，检出限分别为170和16 pg mL -1。该测定法具有操作简便，检测灵敏，靶标灵活，信号读取多样化等优点，为商业应用提供了巨大的机会。
更新日期：2017-09-07
中文翻译：
目标诱导的环状DNA酶的形成，用于比色法和化学发光成像法检测蛋白质生物标志物
提出了一种目标诱导的DNAzyme形成的循环策略，以简便的比色法或化学发光成像读数实现蛋白质生物标志物的简单，灵敏和通用检测。在测定中，靶蛋白被一对DNA标记的抗体（Ab1-DNA1和Ab2-DNA2）识别以形成邻近的复合物，该复合物可与缀合物DNA3 / DNA4杂交以释放富含鸟嘌呤的DNA4，从而形成了G-四链体/血红素辣根过氧化物酶模拟DNAzyme。通过切割DNA3释放邻近的复合物，可以用核酸外切酶III进一步循环利用该过程，从而导致DNA酶的循环形成。G-quadruplex / hemin DNAzyme可以催化H 2 O 2介导的3,3,5,5-四甲基联苯胺的氧化产生从无色到蓝色的颜色变化或增强luminol–H 2 O 2系统的化学发光。因此，可以用裸眼，比色法和化学发光成像读出信号。以癌胚抗原为模型靶标，所提出的测定法显示了比色法和化学发光成像测定法的检测范围为4个数量级，检出限分别为170和16 pg mL -1。该测定法具有操作简便，检测灵敏，靶标灵活，信号读取多样化等优点，为商业应用提供了巨大的机会。
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