Detection of human neutrophil elastase by aptamer affinity capillary electrophoresis coupled with laser-induced fluorescence using specified site fluorescently labeled aptamer

CORC > 生态环境研究中心 > 中国科学院生态环境研究中心 > 环境化学与生态毒理学国家重点实验室

	Detection of human neutrophil elastase by aptamer affinity capillary electrophoresis coupled with laser-induced fluorescence using specified site fluorescently labeled aptamer
	Wang, Hailin; Bai, Yunlong; Zhao, Qiang
刊名	ANALYTICAL AND BIOANALYTICAL CHEMISTRY
	2017-05-21
卷号	409 期号:29 页码:6843-6849
关键词	Human Neutrophil Elastase Aptamer Capillary Electrophoresis Laser-induced Fluorescence Affinity Binding Protein Detection
英文摘要	As a multifunctional serine protease, human neutrophil elastase (HNE) plays critical roles in a variety of physiopathological processes, such as acute lung injury, emphysema, atherosclerosis, and arthritis. The quantification of HNE is important in many applications. In this paper, we report an aptamer affinity capillary electrophoresis coupled with laser-induced fluorescence (CE-LIF) assay for detection of HNE using a tetramethylrhodamine (TMR)-labeled DNA aptamer probe. The affinity complex of HNE and DNA aptamer probe was well separated from the unbound aptamer probe in CE separation based on the difference of electrophoretic mobility. Broad complex peaks appeared due to possible multiple binding. The 45-mer aptamer having TMR labeling on the 40th T base was used as affinity probe, as larger complex peaks were obtained. We investigated the effects of various metal cations (Na+, K+, and Mg2+) in sample buffer on the binding of HNE and the aptamer in CE-LIF analysis. The presence of Na+, K+, or Mg2+ in sample buffer caused a decrease of complex peaks, and Mg2+ showed a larger effect. Under optimized conditions, this aptamer CE-LIF assay enabled the detection of HNE at 0.5 nM. This assay showed good specificity and allowed for detection of HNE spiked in diluted human serum sample.
内容类型	期刊论文
源URL	[http://ir.rcees.ac.cn/handle/311016/39286]
专题	生态环境研究中心_环境化学与生态毒理学国家重点实验室
推荐引用方式 GB/T 7714	Wang, Hailin,Bai, Yunlong,Zhao, Qiang. Detection of human neutrophil elastase by aptamer affinity capillary electrophoresis coupled with laser-induced fluorescence using specified site fluorescently labeled aptamer[J]. ANALYTICAL AND BIOANALYTICAL CHEMISTRY,2017,409(29):6843-6849.
APA	Wang, Hailin,Bai, Yunlong,&Zhao, Qiang.(2017).Detection of human neutrophil elastase by aptamer affinity capillary electrophoresis coupled with laser-induced fluorescence using specified site fluorescently labeled aptamer.ANALYTICAL AND BIOANALYTICAL CHEMISTRY,409(29),6843-6849.
MLA	Wang, Hailin,et al."Detection of human neutrophil elastase by aptamer affinity capillary electrophoresis coupled with laser-induced fluorescence using specified site fluorescently labeled aptamer".ANALYTICAL AND BIOANALYTICAL CHEMISTRY 409.29(2017):6843-6849.