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A new two-mode fluorescence signal amplification strategy for protease activity assay based on graphene oxide

机译:基于石墨烯氧化物的蛋白酶活性测定新的两种荧光信号放大策略

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摘要

A graphene oxide (GO)-based two-mode fluorescence signal amplification assay of protease activity has been established. GO can adsorb the FITC-labeled substrate peptide and quench the fluorescence of FITC. In the presence of the target protein, carboxypeptidase Y (CPY), the FITC-labeled substrate peptide is hydrolyzed by CPY, leading to the turn-on of fluorescence. The fluorescence intensity increases significantly after the hydrolysis. More interestingly, it is even much higher than the fluorescence intensity of the added FITC-labeled substrate peptides. It is deduced that the extraordinary growing of fluorescence intensity is attributed to the hydrolysis also. The strong quenching efficiency of GO significantly improved the signal-to-noise ratio (SNR) of the proposed method for protease analysis. By combining the GO-based fluorescence turn-on with the fluorescence signal amplification induced by hydrolysis, the proposed method obtained higher sensitivity and specificity for CPY activity detection. The detection limit for CPY activity assay is estimated to be 1.0 x 10(-5) U mu L-1. The other proteins, proteases and a complicated matrix cannot disturb the assay of CPY activity.
机译:已经建立了基于蛋白酶活性的氧化物氧化物(GO)的双模荧光信号放大测定。 GO可以吸附FITC标记的底物肽并淬灭FITC的荧光。在靶蛋白的存在下,羧肽酶Y(CPY),FyC标记的底物肽通过CPY水解,导致荧光的开启。水解后荧光强度显着增加。更有趣的是,它甚至高于添加的FITC标记的底物肽的荧光强度。推导出荧光强度的非凡生长归因于水解。淬火效率的强烈淬火效率显着改善了所提出的蛋白酶分析方法的信噪比(SNR)。通过将基于荧光导通与水解诱导的荧光信号相结合,所提出的方法得到了更高的CPY活性检测灵敏度和特异性。 CPY活性测定的检测限估计为1.0×10(-5)U mu L-1。其他蛋白质,蛋白酶和复杂基质不能干扰CPY活性的测定。

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  • 来源
    《RSC Advances》 |2017年第76期|共7页
  • 作者单位

    Hebei Univ Coll Chem &

    Environm Sci Key Lab Analyt Sci &

    Technol Hebei Prov Key Lab Med Chem &

    Mol Diag Minist Educ Baoding 071002 Peoples R China;

    Hebei Univ Coll Chem &

    Environm Sci Key Lab Analyt Sci &

    Technol Hebei Prov Key Lab Med Chem &

    Mol Diag Minist Educ Baoding 071002 Peoples R China;

    Hebei Univ Coll Chem &

    Environm Sci Key Lab Analyt Sci &

    Technol Hebei Prov Key Lab Med Chem &

    Mol Diag Minist Educ Baoding 071002 Peoples R China;

    Hebei Univ Coll Chem &

    Environm Sci Key Lab Analyt Sci &

    Technol Hebei Prov Key Lab Med Chem &

    Mol Diag Minist Educ Baoding 071002 Peoples R China;

    Hebei Univ Med Comprehens Expt Ctr Baoding 071000 Peoples R China;

    Hebei Univ Coll Chem &

    Environm Sci Key Lab Analyt Sci &

    Technol Hebei Prov Key Lab Med Chem &

    Mol Diag Minist Educ Baoding 071002 Peoples R China;

    Hebei Univ Coll Chem &

    Environm Sci Key Lab Analyt Sci &

    Technol Hebei Prov Key Lab Med Chem &

    Mol Diag Minist Educ Baoding 071002 Peoples R China;

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  • 原文格式 PDF
  • 正文语种 eng
  • 中图分类 化学;
  • 关键词

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