Development of indirect competitive fluorescence immunoassay for 2,2 ',4,4 '-tetrabromodiphenyl ether using DNA/dye conjugate as antibody multiple labels

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

	Development of indirect competitive fluorescence immunoassay for 2,2 ',4,4 '-tetrabromodiphenyl ether using DNA/dye conjugate as antibody multiple labels
	Fan, Zi-Yan ; Keum, Young Soo ; Li, Qing-Xiao ; Shelver, Weilin L. ; Guo, Liang-Hong
刊名	JOURNAL OF ENVIRONMENTAL SCIENCES-CHINA
	2012
卷号	24 期号:7 页码:1334－1340
关键词	polybrominated diphenyl ethers fluorescence immunoassay microplate multiple labeling
ISSN号	1001-0742
中文摘要	An indirect competitive fluorescence immunoassay using a DNA/dye conjugate as antibody multiple labels was developed on 96-well plates for the identification and quantification of 2,2',4,4'-tetrabromodiphenyl ether (BDE-47) in aqueous samples. A hapten, 2,4,2'-tribromodiphenyl ether-4'-aldehyde, was synthesized, and was conjugated to bovine serum albumin to form a coating antigen. Specific recognition of the antigen by anti-PBDE antiserum was confirmed by a surface plasmon resonance measurement. In the immunoassay, the coating antigen was adsorbed on a 96-well plate first, and a sample, antiserum and biotinylated goat anti-rabbit secondary antibody were then added and reacted sequentially. A biotinylatecl, double-stranded DNA with 219 base pairs was attached to the secondary antibody by using streptavidin as a molecular bridge. In situ multiple labeling of the antibody was accomplished after addition of a DNA-binding fluorescent dye, SYBR Green I. The working range of the immunoassay for the BDE-47 standard was 3.1-390 mu g/L, with an IC50 value of 15.6 mu g/L. The calculated LOD of the immunoassay is 0.73 mu g/L. The immunoassay demonstrated relatively high selectivity for BDE-47, showing very low cross-reactivity (< 3%) with BDE-15, BDE-153 and BDE-209. With a spiked river water sample containing 50 mu g/L BDE-47, quantification by the immunoassay was 41.9 mu g/L, which compared well with the standard GC-ECD method (45.7 mu g/L). The developed immunoassay provides a rapid screening tool for polybrominated diphenyl ethers in environmental samples.
WOS记录号	WOS:000306581100022
公开日期	2014-11-04
内容类型	期刊论文
源URL	[http://ir.rcees.ac.cn/handle/311016/7780]
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	Fan, Zi-Yan,Keum, Young Soo,Li, Qing-Xiao,et al. Development of indirect competitive fluorescence immunoassay for 2,2 ',4,4 '-tetrabromodiphenyl ether using DNA/dye conjugate as antibody multiple labels[J]. JOURNAL OF ENVIRONMENTAL SCIENCES-CHINA,2012,24(7):1334－1340.
APA	Fan, Zi-Yan,Keum, Young Soo,Li, Qing-Xiao,Shelver, Weilin L.,&Guo, Liang-Hong.(2012).Development of indirect competitive fluorescence immunoassay for 2,2 ',4,4 '-tetrabromodiphenyl ether using DNA/dye conjugate as antibody multiple labels.JOURNAL OF ENVIRONMENTAL SCIENCES-CHINA,24(7),1334－1340.
MLA	Fan, Zi-Yan,et al."Development of indirect competitive fluorescence immunoassay for 2,2 ',4,4 '-tetrabromodiphenyl ether using DNA/dye conjugate as antibody multiple labels".JOURNAL OF ENVIRONMENTAL SCIENCES-CHINA 24.7(2012):1334－1340.