A fluorescence ratiometric protein assay using light-harvesting conjugated polymers

被引:25
作者
An, Lingling [1 ]
Tang, Yanli [1 ]
Wang, Shu [1 ]
Li, Yuliang [1 ]
Zhu, Daoben [1 ]
机构
[1] Chinese Acad Sci, Inst Chem, Key Lab Organ Solids, Beijing Natl Lab Mol Sci, Beijing 100080, Peoples R China
关键词
conjugated polymers; fluorescent ratiometric technique; protein assay;
D O I
10.1002/marc.200600214
中图分类号
O63 [高分子化学(高聚物)];
学科分类号
070305 ; 080501 ; 081704 ;
摘要
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-bis(6'-N,N,N-trimethylammonium)-hexyl-fluorene phenylene]-bromide (PFP-NMe3+) and the negatively charged biotinylated fluorescein probe (Fl-B) were used to detect the target protein streptavidin optically. the strong, electrostatic interactions between PFP-NMe3+ and fluorescein result in efficient FRET from PFP-NMe3+ to fluorescein. In the presence of streptavidin, however, the biotin moiety of Fl-B specifically associates with streptavidin and the fluorescein molecule is burned deeply in the adjacent vacant binding sites. this separates the fluorescein spatially from the PFP-NMe3+ moiety, resulting in inefficient FRET from PFP-NMe3+ to fluorescein. Although a nonspecific protein, such as BSA, shows nonspecific interactions with PFP-NMe3+, it does not affect the fluorescent ratio value of PFP-NMe3+ 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.
引用
收藏
页码:993 / 997
页数:5
相关论文
共 27 条
[1]   Fluorescence superquenching of conjugated polyelectrolytes: applications for biosensing and drug discovery [J].
Achyuthan, KE ;
Bergstedt, TS ;
Chen, L ;
Jones, RM ;
Kumaraswamy, S ;
Kushon, SA ;
Ley, KD ;
Lu, L ;
McBranch, D ;
Mukundan, H ;
Rininsland, F ;
Shi, X ;
Xia, W ;
Whitten, DG .
JOURNAL OF MATERIALS CHEMISTRY, 2005, 15 (27-28) :2648-2656
[2]   Poly(aryleneethynylene)s: Syntheses, properties, structures, and applications [J].
Bunz, UHF .
CHEMICAL REVIEWS, 2000, 100 (04) :1605-1644
[3]   Fluorescent polymeric transducer for the rapid, simple, and specific detection of nucleic acids at the zeptomole level [J].
Doré, K ;
Dubus, S ;
Ho, HA ;
Lévesque, I ;
Brunette, M ;
Corbeil, G ;
Boissinot, M ;
Boivin, G ;
Bergeron, MG ;
Boudreau, D ;
Leclerc, M .
JOURNAL OF THE AMERICAN CHEMICAL SOCIETY, 2004, 126 (13) :4240-4244
[4]   Perturbation of fluorescence by nonspecific interactions between anionic poly(phenylenevinylene)s and proteins: Implications for biosensors [J].
Dwight, SJ ;
Gaylord, BS ;
Hong, JW ;
Bazan, GC .
JOURNAL OF THE AMERICAN CHEMICAL SOCIETY, 2004, 126 (51) :16850-16859
[5]   High-efficiency fluorescence quenching of conjugated polymers by proteins [J].
Fan, CH ;
Plaxco, KW ;
Heeger, AJ .
JOURNAL OF THE AMERICAN CHEMICAL SOCIETY, 2002, 124 (20) :5642-5643
[6]   DNA hybridization detection with water-soluble conjugated polymers and chromophore-labeled single-stranded DNA [J].
Gaylord, BS ;
Heeger, AJ ;
Bazan, GC .
JOURNAL OF THE AMERICAN CHEMICAL SOCIETY, 2003, 125 (04) :896-900
[7]   DNA detection using water-soluble conjugated polymers and peptide nucleic acid probes [J].
Gaylord, BS ;
Heeger, AJ ;
Bazan, GC .
PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA, 2002, 99 (17) :10954-10957
[8]  
GREEN NM, 1990, METHOD ENZYMOL, V184, P51
[9]   Fluorescence-amplifying detection of hydrogen peroxide with cationic conjugated polymers, and its application to glucose sensing [J].
He, F ;
Tang, YL ;
Yu, MH ;
Wang, S ;
Li, YL ;
Zhu, DB .
ADVANCED FUNCTIONAL MATERIALS, 2006, 16 (01) :91-94
[10]   Fluorescent amplifying recognition for DNA G-quadruplex folding with a cationic conjugated polymer: A platform for homogeneous potassium detection [J].
He, F ;
Tang, YL ;
Wang, S ;
Li, YL ;
Zhu, DB .
JOURNAL OF THE AMERICAN CHEMICAL SOCIETY, 2005, 127 (35) :12343-12346