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Application of nano-gold directly bonded with luminol in immunoassay

An immunoassay and direct bonding technology, applied in its application field in immunoassay, can solve the problems such as the lack of research on the application of nano-gold, the difficulty of measuring with actual samples, and the unsatisfactory stability, so as to overcome the trouble of labeling technology, The effect of stabilizing luminous efficiency and reducing luminous efficiency

Active Publication Date: 2010-12-01
UNIV OF SCI & TECH OF CHINA
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

But measuring H 2 o 2 The sensitivity is low, and its detection limit is only 1×10 -7 mol / L H 2 o 2
In addition, the stability of this analytical method is not ideal, and it is difficult to be used for the determination of actual samples
In the above work, the application of luminol directly bonded gold nanoparticles in bioanalysis such as immunoassay was not studied

Method used

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  • Application of nano-gold directly bonded with luminol in immunoassay
  • Application of nano-gold directly bonded with luminol in immunoassay
  • Application of nano-gold directly bonded with luminol in immunoassay

Examples

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Effect test

preparation example Construction

[0058] The preparation method of the nano-gold immunoassay probe directly bonded to luminol of the present invention comprises the following steps:

[0059] (1) 100 mL of 0.01% (w / w) HAuCl 4 Heat the solution to boiling, under the conditions of condensing and refluxing and fully stirring, add 1.5-2.0mL solution containing 0.01mol / L luminol and 0.01mol / L sodium hydroxide, keep boiling and continue to stir and reflux for 30min, then remove The heating source continues to stir for more than 15 minutes to obtain a nano-gold solution with a particle diameter of 14-25nm;

[0060] (2) The synthesized nano-gold solution was dialyzed at normal temperature for 2 days, and the ultrapure water dialyzate was changed every 8 hours to obtain the nano-gold directly bonded to luminol;

[0061] (3) Add 0.5 mL of 1.0 mg / mL antibody to the gold colloid solution dialyzed in the above step (2) with 0.1 mol / L sodium hydroxide to adjust the pH to 8.0, and incubate at room temperature for 30 minutes ...

Embodiment 1

[0114] Embodiment 1. Preparation of the chemiluminescent immunoprobe of the present invention

[0115] (1) 100 mL of 0.01% (w / w) HAuCl 4 Heat the solution to boiling, under the conditions of condensing and refluxing and fully stirring, quickly add 1.5ml of a solution containing 0.01mol / L luminol and 0.01mol / L sodium hydroxide, keep boiling and continue to stir and reflux for 30min, then remove the heating Continue to stir for more than 15 minutes to obtain a nano-gold solution with a particle size of 25nm, cool to room temperature, and store at 4°C in the dark;

[0116] (2) The nano-gold solution synthesized in step (1) was dialyzed at normal temperature for 2 days, and the ultrapure water dialysate was changed every 8 hours. Free luminol and its oxidation products could be fully removed by the dialyzing operation, so that the nano-gold Luminol molecules in gold only exist in the form of bonding with the surface of gold nanoparticles, that is, gold nanoparticles directly bond...

Embodiment 2

[0118] Example 2. Immobilization of Sandwich Immune Complexes on Nanogold Modified Electrodes

[0119] like figure 2 As shown, the immobilization of the sandwich-type immune complex involved in the chemiluminescent immunoassay method of the present invention on the nano-gold modified electrode comprises the following steps:

[0120] (1) Preparation of nano-gold modified electrode:

[0121] A. Put the polished bare gold electrode into 2mmol / L 1,3'-propanedithiol, soak it at room temperature for 20 hours, take it out and wash it with ethanol and ultrapure water to obtain 1,3'-propanedithiol Dithiol-modified gold electrodes;

[0122] B. Drop 60 μL of streptavidin-linked gold nanoparticles on the 1,3′-propanedithiol-modified electrode prepared in step A, put it in the dark at 4°C for 4 hours, take it out and wash it with ultrapure water Washing to obtain a gold electrode modified by streptavidin / nanogold / 1,3'-propanedithiol;

[0123] (2) Immobilization of the double-antibody ...

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Abstract

The invention discloses an application of nano-gold directly bonded with luminol in an immunoassay. The invention is characterized in that the immunoassay probe of a nano-gold directly bonded with luminol comprises antibodies which are labeled by the nano-golds directly bonded with luminol, wherein the nano-gold directly bonded with luminol are prepared through reducing chloroauric acid by the luminol at a single step; a chemiluminescence immunoassay method is based on the immunoassay probe of the nano-gold directly bonded with luminol; and a kit is used for carrying out the immunoassay method. The chemiluminescence immunoassay method of the invention has the advantages of high sensitivity (for instance, the detection limit can reach 1.0pg / mL for detecting human IgG), wide range of linearity, good repeatability, simple operation, low cost and the like, can be applied to detect multi-analyte in various samples and has key application prospect in fields, such as clinical diagnosis and cure, drug analysis, food security detection, environmental monitoring and the like.

Description

technical field [0001] The invention relates to the application of nano-gold directly bonded with luminol, in particular to its application in immunoassay. Background technique [0002] Chemiluminescence is the emission of light during a chemical reaction. As an optical detection method, it does not require a light source, has the advantages of high sensitivity, wide linear range, low background noise, simple instrument, and low price, and has been widely used in immunoassays (Wu, A.H.B.Clin.Chim.Acta 2006 , 369, 119; Zhan, W.; Bard, A.J.Anal.Chem.2007, 79, 459; Yin, X.B.; Qi, B.; Sun; X.P.; Yang; X.R.; Wang; , 3525; Jie, G.F.; Huang, H.P.; Sun, X.1.; Zhu, J.J.Biosens.Bioelectron.2008, 23, 1896; Egashira, N.; Morita, S.; ; Uda, T.Anal.Chem.2008, 80, 4020), has become one of the main clinical analysis methods in many large hospitals. The performance of various chemiluminescent immunoassay methods depends largely on the probes employed. Existing commercial chemiluminescent...

Claims

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Application Information

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IPC IPC(8): G01N33/53G01N33/543G01N33/532G01N21/66G01N33/552
CPCG01N33/54346G01N33/587G01N33/533G01N33/553G01N33/582
Inventor 崔华田大勇
Owner UNIV OF SCI & TECH OF CHINA
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