title: Rapid Immobilization of Oligonucleotides at High Density on Semiconductor Quantum Dots and Gold Nanoparticles.
abstract: Oligonucleotide-coated nanoparticles (NPs) have been used in numerous applications such as bioassays, as intracellular probes, and for drug delivery. One challenge that is confronted in the preparation of oligonucleotide-NP conjugates is derived from surface charge because nanoparticles are often stabilized and made water-soluble with a coating of negatively charged capping ligands. Therefore, an electrostatic repulsion is present when attempting to conjugate oligonucleotides. The result is that the conjugation can be a slow process, sometimes requiring 1 to 2 days to equilibrate at the highest surface density. The effect is compounded by electrostatic repulsion between neighboring oligonucleotide strands on the NP surfaces, which tends to lower the surface density. Herein, we report a novel method that enables conjugation in less than 1 min with a surface density of oligonucleotides up to the theoretical physical limit of occupancy. Negatively charged NPs are first adsorbed onto the surface of positively charged magnetic beads (MBs) to create MB-NP conjugates. Oligonucleotides are subsequently electrostatically adsorbed onto the MB surfaces when added to a suspension of MB-NP conjugates. This creates an oligonucleotide concentration 105to 106greater than in bulk solution in the vicinity of the nanoparticles, resulting in the promotion of the kinetics by over 1000-fold and achieving the maximum density possible for the conjugation reaction.
authors.affiliation: Chemical and Physical Sciences, Univeristy of Toronto Mississauga , Davis Building, 3359 Mississauga Road, Mississauga, Ontario, Canada L5L 1C6.
authors.forename: Abootaleb
authors.initials: A
authors.lastname: Sedighi
authors.affiliation: Chemical and Physical Sciences, Univeristy of Toronto Mississauga , Davis Building, 3359 Mississauga Road, Mississauga, Ontario, Canada L5L 1C6.
authors.forename: Ulrich J
authors.identifier: 0000-0001-8836-7648
authors.initials: UJ
authors.lastname: Krull
mesh_terms: D006046:Gold; D061925:Immobilized Nucleic Acids / Q000737:chemistry*; D053768:Metal Nanoparticles; D009841:Oligonucleotides / Q000737:chemistry*; D045663:Quantum Dots; D012666:Semiconductors
chemical_list: D061925:Immobilized Nucleic Acids; D009841:Oligonucleotides; D006046:Gold