Part-per-trillion level detection of estradiol by competitive fluorescence immunoassay using DNA/dye conjugate as antibody multiple labels
文献类型：期刊论文
| 作者 | Zhu, Shengchao; Zhang, Qin; Guo, Liang-Hong |
| 刊名 | ANALYTICA CHIMICA ACTA
 |
| 出版日期 | 2008-08-22 |
| 卷号 | 624期号:1页码:141-146 |
| 关键词 | estradiol competitive immunoassay fluorescence DNA/dye label |
| 英文摘要 | Fluorescent organic dyes are currently the standard signal-generating labels used in microarray quantification. However, new labeling strategies are needed to meet the demand for high sensitivity in the detection of low-abundance proteins and small molecules. in this report, a long-chain DNA/dye conjugate was used to attach multiple fluorescence labels on antibodies to improve signal intensity and immunoassay sensitivity. Compared with the 30 base-pair (bp) oligonucleotide used in our previous work [Q. Zhang, L.-H. Guo, Bioconjugate Chem. 18 (2007) 1668-1672], conjugation of a 219bp DNA in solution with a fluorescent DNA binder SYBR Green I resulted in more than sixfold increase in signal intensity, consistent with the increase in bp number. In a direct immunoassay for the detection of goat anti-mouse IgG in a mouse IgG-coated 96-well plate, the long DNA conjugate label also produced higher fluorescence than the short one, accompanied by about 15-fold improvement in the detection limit. To demonstrate its advantage in real applications, the DNA/dye conjugate was employed in the competitive immunoassay of 17 beta-estradiol, a clinically and environmentally important analyte. The biotin-terminated DNA was attached to biotinylated anti-estradiol antibody through the biotin/streptavidin/biotin bridge after the immuno-reaction was completed, followed by conjugation with SYBR Green I. The limit of detection for 17 beta-estradiol is 1.9 pg mL(-1), which is 200-fold lower than the assay using fluorescein-labeled antibodies. The new multiple labeling strategy uses readily available reagents, and is also compatible with current biochip platform. It has great potential in the sensitive detection of protein and antibody microarrays. (C) 2008 Elsevier B.V. All rights reserved. |
| WOS记录号 | WOS:000259354900013 |
| 源URL | [http://ir.rcees.ac.cn/handle/311016/21970]  |
| 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 | Zhu, Shengchao,Zhang, Qin,Guo, Liang-Hong. Part-per-trillion level detection of estradiol by competitive fluorescence immunoassay using DNA/dye conjugate as antibody multiple labels[J]. ANALYTICA CHIMICA ACTA,2008,624(1):141-146. |
| APA | Zhu, Shengchao,Zhang, Qin,&Guo, Liang-Hong.(2008).Part-per-trillion level detection of estradiol by competitive fluorescence immunoassay using DNA/dye conjugate as antibody multiple labels.ANALYTICA CHIMICA ACTA,624(1),141-146. |
| MLA | Zhu, Shengchao,et al."Part-per-trillion level detection of estradiol by competitive fluorescence immunoassay using DNA/dye conjugate as antibody multiple labels".ANALYTICA CHIMICA ACTA 624.1(2008):141-146. |
入库方式： OAI收割
来源：生态环境研究中心
浏览0
下载0
收藏0
其他版本
除非特别说明，本系统中所有内容都受版权保护，并保留所有权利。