Fluorescent signal amplification of carbocyanine dyes using engineered viral nanoparticles.
We report enhancement in the fluorescent signal of the carbocyanine dye Cy5 by using an engineered virus as a scaffold to attach >40 Cy5 reporter molecules at fixed locations on the viral capsid. Although cyanine dye loading is often accompanied by fluorescence quenching, our results demonstrate that organized spatial distribution of Cy5 reporter molecules on the capsid obviates this commonly encountered problem. In addition, we observe energy transfer from the virus to adducted dye molecules, resulting in a highly fluorescent viral nanoparticle. We have used this enhanced fluorescence for the detection of DNA-DNA hybridization. When compared with the most often used detection methods in a microarray-based genotyping assay for Vibrio cholerae O139, these viral nanoparticles markedly increased assay sensitivity, thus demonstrating their applicability for existing DNA microarray protocols.
Soto
Carissa M
CM
Center for Bio/Molecular Science and Engineering, Naval Research Laboratory, 4555 Overlook Avenue SW, Washington, DC 20375, USA. cmsoto@cbmse.nrl.navy.mil
Blum
Amy Szuchmacher
AS
Vora
Gary J
GJ
Lebedev
Nikolai
N
Meador
Carolyn E
CE
Won
Angela P
AP
Chatterji
Anju
A
Johnson
John E
JE
Ratna
Banahalli R
BR
D001360:Avidin / Q000737:chemistry; D002232:Carbocyanines / Q000737:chemistry*; D017798:Comovirus / Q000737:chemistry* / Q000235:genetics; D004247:DNA / Q000032:analysis; D015342:DNA Probes / Q000737:chemistry; D004279:DNA, Viral / Q000032:analysis; D005456:Fluorescent Dyes / Q000737:chemistry*; D005818:Genetic Engineering; D008301:Maleimides / Q000737:chemistry; D008958:Models, Molecular; D053758:Nanoparticles / Q000737:chemistry*; D012235:Rhodamines / Q000737:chemistry; D013050:Spectrometry, Fluorescence; D014734:Vibrio cholerae / Q000737:chemistry / Q000235:genetics
D002232:Carbocyanines; D015342:DNA Probes; D004279:DNA, Viral; D005456:Fluorescent Dyes; D008301:Maleimides; D012235:Rhodamines; C085321:cyanine dye 5; C093483:neutravidin; D001360:Avidin; C043592:maleimide; D004247:DNA