Determination of the platelet-derived growth factor BB by a competitive thrombin-linked aptamer-based Fluorometric assay
文献类型：期刊论文
| 作者 | Guo, Limin; Zhao, Qiang |
| 刊名 | MICROCHIMICA ACTA
 |
| 出版日期 | 2016-12 |
| 卷号 | 183期号:12页码:3229-3235 |
| 关键词 | Aptasensor Fluorescence Enzyme label Microplate assay Microtiter plate Fluorogenic enzyme substrate Serine protease DNA probe Affinity binding Dual functional probe Displacement Indirect analysis |
| 英文摘要 | The authors describe a competitive aptamer based assay for detection of the platelet-derived growth factor BB (PDGF-BB; used as a model protein). The assay is making use of thrombin (a serine protease) as an enzyme label for reporting signals. It is taking advantage of a highly selective aptamer and of the fairly specific enzymatic activity of thrombin in terms of cleaving artificial fluorogenic peptide substrates. In a first step, the surface of wells of microplates is coated with PDGF-BB. On addition of a sample containing PDGF-BB, free and bound PDGF-BB compete with each other for binding to a DNA probe that consists of an aptamer sequence for PDGF-BB and a 29-mer aptamer sequence for thrombin. After washing, thrombin is added and will attach to the DNA probe that bound to the PDGF-BB on the microplates. Following addition of a fluorogenic peptide substrate, the bound thrombin will catalyze the cleavage of the substrate to generate a fluorescent product whose fluorescence intensity is measured at excitation/emission wavelengths of 370/440 nm. Fluorescence intensity decreases with increasing PDGF-BB concentration in the sample because less thrombin will bind to the PDGF-BB coated surface of the microplate. Under optimal conditions, PDGF-BB can be quantified in the 0.125 to 3 nM concentration range. This assay was successfully applied to the determination of PDGF-BB in spiked 100-fold diluted human serum. |
| 收录类别 | SCI |
| 源URL | [http://ir.rcees.ac.cn/handle/311016/35834]  |
| 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 | Guo, Limin,Zhao, Qiang. Determination of the platelet-derived growth factor BB by a competitive thrombin-linked aptamer-based Fluorometric assay[J]. MICROCHIMICA ACTA,2016,183(12):3229-3235. |
| APA | Guo, Limin,&Zhao, Qiang.(2016).Determination of the platelet-derived growth factor BB by a competitive thrombin-linked aptamer-based Fluorometric assay.MICROCHIMICA ACTA,183(12),3229-3235. |
| MLA | Guo, Limin,et al."Determination of the platelet-derived growth factor BB by a competitive thrombin-linked aptamer-based Fluorometric assay".MICROCHIMICA ACTA 183.12(2016):3229-3235. |
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。