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A covalent labeling method of colloidal gold for rapid detection

A labeling method and technology of colloidal gold particles, which can be used in measurement devices, instruments, scientific instruments, etc., can solve the problems of false positive or signal light color rendering, instability, separation, etc., to achieve selective identification enhancement, improve stability, The effect of increasing sensitivity

Active Publication Date: 2016-06-15
ZHUHAI LIVZON DIAGNOSTICS
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0004] Since the electrostatic and hydrophobic interactions are relatively weak forces, the gold-labeled complex is very unstable during the chromatographic surge process, and separation will occur, resulting in problems such as false positives or light color of the signal.

Method used

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  • A covalent labeling method of colloidal gold for rapid detection
  • A covalent labeling method of colloidal gold for rapid detection
  • A covalent labeling method of colloidal gold for rapid detection

Examples

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preparation example Construction

[0022] (1) Preparation of colloidal gold:

[0023] a. Prepare colloidal gold particles with a small particle size of 10-15nm by traditional sodium citrate reduction method: heat and stir 0.01% (w / v) chloroauric acid solution to boiling, quickly add reducing agent trisodium citrate solution, wait After the color of the solution turns bright red until it stops changing, continue heating and boiling for 5 minutes, stop stirring, and cool to room temperature for later use;

[0024] b. Method (1): The technical route is as follows: figure 1 As shown, use Traut's reagent to react with amino groups on antibodies or proteins to introduce sulfhydryl groups, so as to form Au-S coordination bonds with gold nanoparticles: dilute small-sized colloidal gold particles at a certain concentration as a seed solution and place them in a three-necked flask During the process, stir at room temperature, and slowly and uniformly drop the chloroauric acid solution and the weak reducing agent ascorbi...

example 1

[0034] 1. Dilute 1% (w / v) chloroauric acid into 100mL of 0.01% solution with secondary deionized purified water, heat chloroauric acid to boiling, quickly add 4.0mL 1% (w / v) trisodium citrate Solution, until the color of the solution turns bright red until no longer changes, continue to heat and boil for 5 minutes, cool to room temperature and add purified water to make the volume to 100mL, that is, colloidal gold particles with small particle size are obtained;

[0035] 2. Method (I): Take 30mL of small particle size colloidal gold and dilute it to 100mL with deionized purified water in a three-neck flask, add 2.0mL1% (w / v) chloroauric acid dropwise through a feeding tube, and control the dropping rate to 0.8mL / min; add 1.2mL of ascorbic acid solution dropwise into another feeding tube, and control the dropping rate to 0.5mL / min. After the color is stable, scan the particle size distribution and size with a UV-visible spectrophotometer to obtain colloidal gold particles of ab...

example 2

[0041] 1. Dilute 1% (w / v) chloroauric acid into 100mL of 0.01% solution with secondary deionized purified water, heat chloroauric acid to boiling, quickly add 4.0mL 1% (w / v) trisodium citrate Solution, until the color of the solution turns bright red until no longer changes, continue to heat and boil for 5 minutes, cool to room temperature and add purified water to make the volume to 100mL, that is, colloidal gold particles with small particle size are obtained;

[0042] 2. Method (I): Take 30mL of small particle size colloidal gold and dilute it to 100mL with deionized purified water in a three-neck flask, add 2.0mL1% (w / v) chloroauric acid dropwise through a feeding tube, and control the dropping rate to 0.8mL / min; add 2.5mL of hydroxylamine hydrochloride solution dropwise to the other feeding tube, and control the dropping rate to 0.6mL / min. After the color is stable, scan the particle size distribution and size with a UV-visible spectrophotometer to obtain colloidal gold p...

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Abstract

The invention provides a labeling method for immunological detection of nanometer colloidal gold. The covalent labeling method comprises the following steps: preparing a gold nanoparticle with a small particle diameter through a sodium citrate reduction method, using the prepared gold nanoparticle with the small particle diameter as seed gold, and slowly and dropwise adding chloroauric acid and a weak reducing agent at a constant speed so as to prepare a colloidal gold particle with a large particle diameter; treating an antibody to be labeled or protein with a Traut's reagent so as to combine the treated antibody or the treated protein with the prepared colloidal gold in a coupling manner; or, using the prepared gold nanoparticle with the small particle diameter as the seed gold, slowly and dropwise adding the chloroauric acid and a carboxylic acid reducing agent containing sulfhydryl at a constant speed so as to prepare a colloidal gold particle with the large particle diameter and the sulfhydryl on the surface, activating the prepared colloidal gold particle with a cross-linking agent so as to obtain a functional colloidal gold particle, and combining the treated antibody or the treated protein with the prepared colloidal gold particle in the coupling manner so as to form a gold labelled antibody protein compound. Through the use of the covalent labeling method disclosed by the invention, the stability of the gold labelled antibody protein compound in a complex medium is improved, and the selective recognition to objective protein is greatly enhanced, so that the sensitivity of a gold labelled test paper strip is relatively improved.

Description

technical field [0001] The invention relates to the field of biological detection, in particular to a covalent labeling method of nano colloidal gold for rapid detection. Background technique [0002] Colloidal gold is made from chloroauric acid (HAuCl 4 ) is reduced to gold atoms under the action of reducing agents (such as white phosphorus, ascorbic acid, sodium citrate, tannic acid, etc.), and the obtained gold atoms will adsorb and aggregate into gold atom clusters and further form gold nanoparticles, which are due to electrostatic repulsion. Dispersed in solution to become a stable colloidal state. Colloidal gold labeling is essentially a coating process in which polymers such as proteins are adsorbed to the surface of colloidal gold particles. Part of the adsorption mechanism is that colloidal gold is negatively charged in a weak alkaline environment, and the positively charged groups of proteins are due to electrostatic adsorption. formed combination. Proteins cont...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): G01N33/532
CPCG01N33/532
Inventor 曾敏霞李重阳朱越谭
Owner ZHUHAI LIVZON DIAGNOSTICS
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