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Method of manipulating the surface density of functional molecules on nanoparticles

A technology of functional molecules and nanoparticles, which is applied in the field of functional molecules to modify nanoparticles, can solve the problems of DNA density increase, obstruction, and inability to solve well, and achieve the effect of simple method and good cost-effectiveness

Active Publication Date: 2011-03-30
THE HONG KONG UNIV OF SCI & TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, due to the hindering effect of the BSPP layer on the particle surface, DNA density is difficult to increase by this method
[0010] It can be seen from the above that in order to achieve effective regulation of the DNA surface density in a large range from low to high, the above two methods themselves cannot solve the problem very well.
So far, there is still a lack of a unified and rapid method to achieve this control purpose to meet the requirements of different applications for the modification density of nanoparticles.

Method used

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  • Method of manipulating the surface density of functional molecules on nanoparticles
  • Method of manipulating the surface density of functional molecules on nanoparticles
  • Method of manipulating the surface density of functional molecules on nanoparticles

Examples

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Embodiment 1

[0058] Manipulating the binding density of 103bp thiolated double-stranded DNA (103bp-dsDNA) on the surface of 13nm gold nanoparticles

[0059] The 103bp-dsDNA was prepared by performing polymerase chain reaction on the M13 phage vector with thiol primers (thiol reverse primer: 5'-thiol-C6-CAGGAAACAGCTATGAC-3' and forward primer: 5'-GTAAAACGACGGCCAG-3') ( obtained by PCR) amplification, and the PCR product was further used in a kit (PCRquick-spin TM PCRProductPurificationKit) purification. The final concentration of 103bp-dsDNA can be obtained by measuring the absorbance of the sample at a wavelength of 260nm.

[0060] At the same time, 1100 μL of citrate-stabilized 13nm gold nanoparticles was mixed with ATP at a molar ratio of 1:1000 and incubated for 15 minutes, and then 10 mM sodium phosphate buffer at pH 8.0 was added and incubated for another 15 minutes. Divide the incubated mixed solution into 11 equal parts, and then adjust the sodium chloride concentration of 5 parts...

Embodiment 2

[0064] Preparation of low-density DNA-modified gold nanoparticles to assemble gold nanoparticle dimer or trimer nanostructures

[0065] The two complementary thiol DNA strands (thiol-T30, 5'-TTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTT-C3-thiol-3'; and thiol-A30, 5'-AAAAAAAAAAAAAAAAAAAAAAAAAAAAAA-C3-thiol-3') were prepared according to the procedures similar to those in Example 1. The steps described above are combined with gold nanoparticles, except that the molar ratio of thiolated DNA to gold nanoparticles is adjusted to 120:1, and the introduction time of thiolated oligoethylene glycol is 5 minutes ( Figure 5 Medium (4 and 12)), 10 minutes ( Figure 5 Medium (5 and 13)), 15 minutes ( Figure 5 Medium (6 and 14)), 30 minutes ( Figure 5 Medium (7 and 15)) and overnight ( Figure 5 (8 and 16)), two groups of parallel salt concentrations were 0mM ( Figure 5 In (b)) and 50mM ( Figure 5 middle (c)).

[0066] Two gold nanoparticles modified with complementary DNA strands were hybri...

Embodiment 3

[0068] Preparation of DNA / DNA or DNA / peptide co-modified gold nanoparticles

[0069] Preparation method of DNA / DNA co-modified gold nanoparticles:

[0070] Using similar steps as in Example 1, the two DNA strands (thiol-T5, 5'-TTTTTT-C3-thiol-3; and thiol-T30, 5'-TTTTTTTTTTTTTTTTTTTTTTTTTTTTTT-C3-thiol-3') were sequentially Different densities were bound to gold nanoparticles to obtain the complex 1( Image 6 in (1 and 2)). Among them, low-density thiol-T30 was first incubated with gold nanoparticles at a molar ratio of 50:1 in a buffer containing 0 mM sodium chloride for 15 minutes, and then centrifuged (13,200 rpm, 20 minutes) to remove unreacted substances; after that, the high-density thiol-T5 was incubated with gold nanoparticles at a molar ratio of 250:1 in a buffer containing 0.1M sodium chloride for 30 minutes, and finally added thiol oligoethylene glycol and incubated for another 15 minutes to end reaction. Another DNA / DNA co-modified gold nanoparticle conjugate 2...

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Abstract

Provided herein is a method for manipulating the surface density of functional molecules conjugated to nanoparticles, which method including incubating nanoparticles with nucleotides to form nucleotide-coated nanoparticles, adjusting buffer and salt concentration of the conjugation media, adding thiolated molecules in the conjugation media to incubate with the nucleotie-coated nanoparticles, and adding thiolated oligo(ethylene glycol) in the conjugation media to cease the conjugation process of thiolated molecules to nanoparticles. The method is simple, efficient and cost effective, and the surface density of functional molecules can be quickly manipulated in a wide range for various applications, such as biosensing, molecular diagnostics, nanomedicine, and nano-assembly.

Description

[0001] This application claims priority to US Provisional Patent Application No. 61 / 272,160, filed August 24, 2009, entitled "Methods for Controlling Density of Functional Molecules on Nanoparticle Surfaces." This provisional patent application is incorporated herein by reference in its entirety. technical field [0002] The present invention relates to a technology for modifying nanoparticles with functional molecules, in particular to methods for combining sulfhydrylated or phosphorothioated, synthetic or natural nucleic acids (such as DNA) or peptides with nanoparticles, and using the functionalized Nanoparticles as a method for the detection of biomolecules. Background technique [0003] Nanoparticles, especially those of noble metals such as gold, are well known for having physicochemical properties that are related to particle size. Nanoparticles modified by thiol molecules have been widely used in various fields of molecular diagnosis, nanomedicine and nanotechnology...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07H21/04C09K11/02C07K2/00G01N27/447
CPCG01N33/54346B82Y30/00B82Y40/00
Inventor 邢怡铭赵文婷
Owner THE HONG KONG UNIV OF SCI & TECH
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