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A kind of nano-gold composite and its preparation and application

A composite, nano-gold technology, applied in the field of biotechnology and biological detection, can solve the problems of reducing the stability of nano-gold particles, sample contamination, and high cost

Active Publication Date: 2017-05-10
SHANGHAI INST OF APPLIED PHYSICS - CHINESE ACAD OF SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

Applying this method to prepare DNA-nanogold complex requires chemical modification of DNA, the operation process is very complicated, and the cost is high. Unwanted disulfide bonds may be formed during the operation, or there may be problems such as sample contamination.
The common disadvantage of this method is that due to the non-specific adsorption of gold nanoparticles on the surface of incompletely functionalized gold nanoparticles, the adsorption of DNA on the surface of gold nanoparticles presents a lodging state; in addition, although this method carries a high density of DNA, it is not effective in highly functionalized On the gold nanoparticles, the high density of DNA makes the target detection molecules difficult to access, which affects the detection efficiency
[0004] The use of competing small molecules, such as mercaptohexanol (MCH), can improve hybridization efficiency by displacing the DNA adsorbed on the gold surface and forcing the DNA sequence to remain upright, but the number of competing small molecules used in this method and the reaction The control of AuNPs is very complicated, and the introduction of competing small molecules will reduce the stability of AuNPs

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  • A kind of nano-gold composite and its preparation and application
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[0132] The present invention also provides a preparation method of the nano-gold complex: the functional molecule is contacted with the nano-gold particle, so that the functional molecule is bound to the nano-gold particle through the oligonucleotide binding region rich in adenine nucleotide or its analogue. surface, thereby forming nano-gold complexes.

[0133] In a preferred example of the present invention, steps are included: mixing functional molecules with gold nanoparticles to obtain a mixed solution of functional molecules and gold nanoparticles; The connection of the nucleotide binding region and the surface of the nano-gold particle obtains a solution system containing the nano-gold complex; and the nano-gold complex is separated from the solution. Preferably, it also includes the step of washing and / or verifying the performance of the gold nanocomposite. Those skilled in the art can choose conventional aging agents, such as sodium chloride solution, potassium chlor...

Embodiment 1

[0161] Assembly of the gold nanocomposites

[0162] First, mix the gold nanoparticles and the functional molecules containing the A-rich oligonucleotide binding region in a certain ratio (DNA / nano-gold: 200:1), gently blow to mix, and incubate at room temperature for 16 hours, then add 1MPBS (1M NaCl / 0.1M PB, pH 7.0) to make the final concentration 0.1M PBS (add 5-6 times, 5min / time interval), place at room temperature for 16h, centrifuge at 12000r, 4°C, 20min, and discard Add an equal amount of 0.1M PBS (0.1M NaCl / 10mM PB, pH 7.0) to wash off three times, suspend in the hybridization solution (0.3M NaCl / 10mM PB, pH 7.0), and store in a refrigerator at 4°C for a long time. figure 1 Schematic diagrams of functional molecules and gold nanocomposites are shown.

Embodiment 2

[0164] Stability of DNA-nanogold complexes containing polyA-binding domains in electrolyte solutions

[0165] In order to determine that the oligonucleotide DNA can be stably connected to the surface of gold nanoparticles through the polyA binding region in its structure, this embodiment takes three kinds of DNA samples: (a) no poly adenine nucleotide (polyA) in the DNA (b) binding region A10 composed of polyadenine nucleotides in (b) DNA, (c) sulfhydryl group modified at the end of DNA, the transition region in the above DNA is T5, and the functional regions are all ATgAT gTTCg TTgTg . The above three kinds of DNA were respectively combined with 10nm nano-gold particles to form DNA-nano-gold complexes, and their stability in electrolyte solutions with different ionic strengths was compared.

[0166] Experimental principle: Nanogold with a particle size of 10nm dispersed in the solution is red. After the nanogold is aggregated, the color will be blue-shifted due to the surfac...

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Abstract

The invention relates to a nanometer gold complex, as well as a preparation method and an application of the nanometer gold complex. Specifically, the nanometer gold complex provided by the invention comprises nanometer gold particles and a functional molecule, wherein the functional molecule is bound to the surfaces of the nanometer gold particles through an oligonucleotide binding region which is rich in adenine nucleotide or the analogue of the adenine nucleotide, and the functional molecule comprises the oligonucleotide binding region which is rich in the adenine nucleotide or the analogue of the adenine nucleotide, a functional region and a transition area located between the binding region and the functional region. The invention also provides the preparation method and the application of the nanometer gold complex.

Description

technical field [0001] The invention belongs to the field of biotechnology and biodetection, in particular, the invention relates to a nano-gold composite and its preparation and application. Background technique [0002] Nano-gold composite is a commonly used biological nanomaterial, which is composed of functional molecules connected to nano-gold particles in a certain way, and has a wide range of applications in biological diagnosis and other aspects. [0003] DNA-nanogold complex is a common form of nanogold complex. Usually, this DNA-nanogold complex is obtained by the self-assembly of sulfhydryl-modified DNA on the gold surface through gold-sulfur bonds. Applying this method to prepare DNA-nanogold complex requires chemical modification of DNA, which is very complicated and costly, and unnecessary disulfide bonds may be formed during the operation, or there may be problems such as sample contamination. The common disadvantage of this method is that due to the non-spe...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12Q1/68C12Q1/44G01N33/53A61K45/00A61P43/00
CPCA61K45/00C12Q1/6816G01N33/54346C12Q2563/137C12Q2563/155C12Q2521/337
Inventor 樊春海裴昊黄庆
Owner SHANGHAI INST OF APPLIED PHYSICS - CHINESE ACAD OF SCI
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