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Preparation and use of novel nano-Au composite substrate NCS

A nano-gold and composite bottom technology, applied in biological testing, material inspection products, etc., can solve the problems of easy agglutination, different particle sizes, and inability to directly catalyze, so as to improve the detection rate, simplify the operation steps, and achieve high sensitivity. The effect of visual inspection

Active Publication Date: 2009-07-01
INST OF RADIATION MEDICINE ACAD OF MILITARY MEDICAL SCI OF THE PLA
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Problems solved by technology

[0003] Colloidal gold has some inherent disadvantages: different particle sizes, easy agglutination, electrostatic adsorption with biomolecules, unstable labeling and high false positive rate, etc.
The disadvantage of this method is that the detection process is relatively cumbersome. This is because: experiments have shown that HRP cannot directly catalyze the deposition of gold nanoparticles linked by tyramide, but must undergo a two-step reaction to lead to the deposition of gold nanoparticles, that is, HRP first catalyzes the biotinylated tyrosine Amine reaction and subsequent conjugation of biotin to streptavidin-gold nanoparticles

Method used

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  • Preparation and use of novel nano-Au composite substrate NCS
  • Preparation and use of novel nano-Au composite substrate NCS
  • Preparation and use of novel nano-Au composite substrate NCS

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Embodiment 1

[0023] Implementation Example 1, Preparation of Nanogold Composite Substrate NCS and Application in Biochip Detection

[0024] 1. Preparation and purification of NCS

[0025] 1) Preparation:

[0026] Dissolve 10 nmol of Nanogold-NHS (purchased from Nanoprobes Inc, Yaphank NY, USA) in 100 μL of DMSO, then add 400 μL of distilled water and mix well; take 100-200 nmol of DAB (purchased from Sigma-Aldrich, St. Dissolve in 200 μL DMSO first, then add 300 μL HEPES (PH=7.5) and mix well; mix the above two solutions, adjust the pH to 7.8-8.0 with 0.1M NaOH solution, and react at room temperature for 4 hours in the dark, during which every 20-30 minutes vigorously Mix to fully react.

[0027] 2) Purification:

[0028] The reaction mixture was filtered with Sephadex-G25 gel chromatography, and 2×PBS was used as the dilution buffer to collect the light yellow component that first flowed out from the column, which was the nano-gold composite substrate NCS; or it was separated by ultraf...

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Abstract

The invention relates to a preparation method of novel nanometer gold composite substrate NCS and an application, wherein the nanometer gold composite substrate NCS is a compound formed by reacting the NHS group of one molecule of Nanogold-NHS and one amino of one molecule of 3, 3', 4, 4'-quadr-aminobiphenyl (DAB) to form amide bond. The preparation method comprises: dissolving and mixing Nanogold-NHS with DAB of the amount over of 10 to 20 times; reacting for 3 to 5h under room temperature without light; separating and purifying the product via Sephadex-G25 gel chromatography or Amicon Centricon-3 ultra-filtration centrifugal tube. The nanometer gold composite substrate NCS under the catalysis of HRP can induce the direct and massive deposition of nanometer gold particles, thereby being applied in high sensitivity detection of biological chip and the like and the research of novel nanometer gold immunity diagnosis agents.

Description

technical field [0001] The invention relates to the preparation of a new nano-gold composite substrate NCS and its application in the fields of molecular detection and the like. Background technique [0002] Nanogold originally refers to tiny gold particles with a diameter of 1-100nm, and generally exists in the form of a sol dispersed in water, so it is usually called colloidal gold. Colloidal gold has unique physical and chemical properties such as surface effect, small size effect, macroscopic quantum tunneling effect, and good biocompatibility, and is very suitable for labeling and detection of biological macromolecules. Colloidal gold has been used in biological detection research for nearly a century. The emergence of immuno-gold-silver staining (IGSS) technology in the early 1980s is a milestone in the application of colloidal gold in biological detection. This technique first labels the antibody or antigen to be tested with colloidal gold, and then introduces silver...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/52
Inventor 王升启戚红卷陈苏红
Owner INST OF RADIATION MEDICINE ACAD OF MILITARY MEDICAL SCI OF THE PLA
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