Sensitive cell apoptosis assay based on caspase-3 activity detection with graphene oxide-assisted electrochemical signal amplification
Sensitive cell apoptosis assay based on caspase-3 activity detection with graphene oxide-assisted electrochemical signal amplification
复制标题
基于氧化石墨烯辅助电化学信号放大的 caspase-3 活性检测的敏感细胞凋亡测定
DOI：
10.1016/j.bios.2015.02.007
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发表时间：
2015-06-15
影响因子：
12.6
通讯作者：
Yin, Yongmei
中科院分区：
文献类型：
--
作者：
Chen, Hongxia;Zhang, Jiangjiang;Yin, Yongmei
本文报道了一种利用电化学技术检测细胞凋亡的新方法。在这项研究中，caspase-3的活性，这是检测与微分加伏安法（DPV）作为一种替代传统的光谱法的方法，被用作细胞凋亡的指标，而乙酰化的肽Ac-GGHDEVDHGGGC被用作阻断底物。在酪蛋白酶-3的存在下，封闭肽的水解可能释放出活性胺基，这些胺基可以与氧化石墨烯共价结合。因此，电活性亚甲基蓝分子可以通过π-π堆积和静电相互作用进一步附着到电极表面。该方法灵敏度高，检测限低至0.06 pg/mL，为细胞凋亡的检测提供了一种新的方法。并成功地将此方法应用于人肺癌A549细胞诱导凋亡后的细胞凋亡检测。(C)2015 Elsevier B. V.版权所有。
This paper reports a novel approach for the simple assays of cell apoptosis using electrochemical technique. In this study, caspase-3 activity, which was detected with differential plus voltammetry (DPV) as an alternative to conventional spectrometry approach, was employed as an indicator of cell apoptosis and, while an acetylated peptide Ac-GGHDEVDHGGGC was used as the blocked substrate. In the presence of casepase-3, the hydrolysis of blocked peptide might release active amine groups, which could covalently conjugate with graphene oxide. Therefore, electroactive methylene blue molecules could be further attached to the electrode surface through pi-pi stacking and electrostatic interactions. Using this proposed new method, a very sensitive detection of caspase-3 could be achieved with a low detection limit of 0.06 pg/mL, and a new method for sensitive detection of cell apoptosis was developed. Moreover, we have successfully used this new method to detect cell apoptosis with human pulmonary carcinoma A549 cell after apoptosis inducing. (C) 2015 Elsevier B.V. All rights reserved.