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Kelleher, SM,Nooney, RI,Flynn, SP,Clancy, E,Burke, M,Daly, S,Smith, TJ,Daniels, S,McDonagh, C
Multivalent linkers for improved covalent binding of oligonucleotides to dye-doped silica nanoparticles
WOS: 5 ()
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bioconjugation dipodal silanes high brightness silica nanoparticles 8-arm polyethylene glycol hybridization assays miR451 COLLOIDAL STABILITY BIDENTATE LIGANDS DNA MICROARRAYS SURFACE OPTIMIZATION FLUORESCENCE SPECTROSCOPY CHIP DRUG SIZE
This paper describes the fabrication of oligonucleotide-coated Cy5-doped silica nanoparticles using a combination of multivalent linkers and their use in surface-based DNA sandwich hybridization assays. Dipodal silane is introduced as a means to fabricate amine-coated silica nanoparticles and its advantages compared to monopodal silanes are discussed. The use of dipodal silane in conjunction with three different polymer linkers (oxidized dextran, linear and 8-arm polyethylene glycol (PEG)) to immobilize single-stranded DNA to Cy5-doped nanoparticles is investigated and dynamic light scattering measurements and Fourier transform infrared spectroscopy are used to follow the progression of the functionalization of the nanoparticles. We observe a significant improvement in the binding stability of the single-stranded DNA when the dipodal silane and 8-arm PEG are used in combination, when compared to alternative conjugation strategies. Both 8mer and 22mer oligonucleotides are securely conjugated to the high-brightness nanoparticles and their availability to hybridize with a complementary strand is confirmed using solution-based DNA hybridization experiments. In addition, a full surface-based sandwich assay demonstrates the potential these nanoparticles have in the detection of less than 500 femtomolar of a DNA analogue of micro RNA, miR-451.
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