A Fluorescence Ratiometric Protein Assay Using Light‐Harvesting Conjugated Polymers
A Fluorescence Ratiometric Protein Assay Using Light‐Harvesting Conjugated Polymers
复制标题
使用光捕获共轭聚合物进行荧光比率蛋白质测定
DOI：
10.1002/marc.200600214
复制
发表时间：
2006
影响因子：
4.6
通讯作者：
Daoben Zhu
中科院分区：
文献类型：
--
作者：
Lingling An;Yanli Tang;Shu Wang;Yuliang Li;Daoben Zhu
A highly selective protein assay was created which combines the fluorescent ratiometric technique based on FRET with the light-harvesting properties of conjugated polymers. The cationic poly((9,9-bi.s(6'-WV,A'-trimethylammonium)-hexyl)-tluorene phenylene bromide (PFP-NMe + 3 ) and the negatively charged biotinylated fluorescein probe (Fl-B) were used to detect the target protein streptavidin optically. The strong electrostatic interactions between PFP-NMef and fluorescein result in efficient FRET from PFP-NMe + 3 to fluorescein. In the presence of streptavidin, however, the biotin moiety of Fl-B specifically associates with streptavidin and the fluorescein molecule is buried deeply in the adjacent vacant binding sites. This separates the fluorescein spatially from the PFP-NMe + 3 moiety, resulting in inefficient FRET from PFP-NMe + 3 to fiuorescein. Although a nonspecific protein, such as BSA. shows nonspecific interactions with PFP-NMe + 3 , it does not fleet the fluorescent ratio value of PFP-NMe + 3 to fluorescein. Hence, the charged neutral complex of two oppositely charged conjugated polymers can eliminate the nonspecific interactions, and thus optimize their application in protein assays.
