A competitive thrombin-linked aptamer assay for small molecule: aflatoxin B-1

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

	A competitive thrombin-linked aptamer assay for small molecule: aflatoxin B-1
	Wang, Chao 1; Zhao, Qiang 1
刊名	ANALYTICAL AND BIOANALYTICAL CHEMISTRY
	2019-10-01
卷号	411 期号:25 页码:6637-6644
关键词	Aptamer Thrombin Aflatoxin B-1 Enzyme Fluorescence Absorbance
ISSN号	1618-2642
英文摘要	We described a competitive thrombin-linked aptamer assay for small molecule, using aflatoxin B-1 (AFB(1)) as a model, taking advantage of aptamer affinity binding and enzymatic activity of thrombin. We designed a dual functional DNA probe that contained the aptamer sequence for thrombin and the aptamer sequence for AFB(1). Thrombin was labeled on the DNA probe by affinity binding between thrombin and anti-thrombin aptamer. This thrombin-labeled DNA probe was attached on AFB(1)-bovine serum albumin conjugate (BSA-AFB(1)) coated on a microplate through the affinity interaction between AFB(1) and anti-AFB(1) aptamer. The thrombin attached on the microplate catalyzed the cleavage of peptide substrate into detectable product, generating signal for detection. In the presence of AFB(1), free AFB(1) competed with BSA-AFB(1) in the binding to the limited amount of DNA probe, leading to signal decrease. Detection of AFB(1) was achieved by measuring the signal change. Under optimized conditions, AFB(1) was successfully detected in the range from 0.5 nM to 1 mu M when fluorogenic peptide substrate of thrombin and fluorescence analysis were applied. The use of chromogenic peptide substrate in the assay allowed the detection of AFB(1) ranging from 0.5 to 125 nM by simple absorbance analysis. The thrombin-linked aptamer assay showed good selectivity towards AFB(1), and enabled the detection of AFB(1) spiked in diluted beer and corn flour. This TLAA strategy extends the analytical application of thrombin and aptamers in detection of small molecules.
内容类型	期刊论文
源URL	[http://ir.rcees.ac.cn/handle/311016/42491]
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
作者单位	1.Chinese Acad Sci, Res Ctr Ecoenvironm Sci, State Key Lab Environm Chem & Ecotoxicol, Beijing 100085, Peoples R China 2.Univ Chinese Acad Sci, Beijing 100049, Peoples R China
推荐引用方式 GB/T 7714	Wang, Chao,Zhao, Qiang. A competitive thrombin-linked aptamer assay for small molecule: aflatoxin B-1[J]. ANALYTICAL AND BIOANALYTICAL CHEMISTRY,2019,411(25):6637-6644.
APA	Wang, Chao,&Zhao, Qiang.(2019).A competitive thrombin-linked aptamer assay for small molecule: aflatoxin B-1.ANALYTICAL AND BIOANALYTICAL CHEMISTRY,411(25),6637-6644.
MLA	Wang, Chao,et al."A competitive thrombin-linked aptamer assay for small molecule: aflatoxin B-1".ANALYTICAL AND BIOANALYTICAL CHEMISTRY 411.25(2019):6637-6644.