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Lateral flow immunochromatographic assay test paper based on gold magnetic nano-enzyme immune probe as well as preparation method and application of lateral flow immunochromatographic assay test paper

An immunochromatographic detection, gold magnetic nanotechnology, applied in the field of biological analysis, can solve the problems of limiting catalytic effect, catalytic activity influence, etc., and achieves the effect of simple process, low cost, and easy scale production.

Active Publication Date: 2020-08-28
SHAANXI UNIV OF SCI & TECH
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  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, common improved methods include the use of fluorescence, chemiluminescence or magnetic quantitative analysis of detection results, but these analysis methods require specific equipment and professionals to implement the detection
In addition, the catalytic activity of natural enzymes is easily affected by the use environment, which limits the catalytic effect in specific use

Method used

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  • Lateral flow immunochromatographic assay test paper based on gold magnetic nano-enzyme immune probe as well as preparation method and application of lateral flow immunochromatographic assay test paper
  • Lateral flow immunochromatographic assay test paper based on gold magnetic nano-enzyme immune probe as well as preparation method and application of lateral flow immunochromatographic assay test paper

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preparation example Construction

[0039] The invention discloses a method for preparing a lateral flow immunochromatography test paper based on a gold magnetic nano-enzyme immunoprobe, comprising the following steps:

[0040] (1) Fe 3 o 4 / Au Composite Nanoparticles Preparation

[0041] Weigh 100-150 mg of Fe with a particle size of about 20-200 nm 3 o 4 For nanoparticles, add 0.1-0.5g of sodium citrate and 1-5mL of chloroauric acid solution with a concentration of 24mmol / L, dilute to 100-200mL with ultrapure water, and heat the reaction at 100-120°C for 30-60min, namely Available Fe 3 o 4 / Au composite nanoparticles.

[0042] (2) Preparation of gold magnetic nano-enzyme immunoprobe

[0043] By physical adsorption method or chemical method, Fe 3 o 4 Au / Au composite nanoparticles were combined with the first recognition antibody of the target molecule to prepare gold magnetic nano-enzyme immunoprobes.

[0044] The specific method is as follows: the step of preparing the immune probe by chemical method...

Embodiment 1

[0060] Lateral Flow Immunochromatographic Detection of hCG Antigen

[0061] (1) Fe 3 o 4 / Au Composite Nanoparticles Preparation

[0062] Weigh 100mg of Fe with a particle size of about 20nm 3 o 4 For nanoparticles, add 0.1g sodium citrate and 1mL chloroauric acid solution with a concentration of 24mmol / L, dilute to 100mL with ultrapure water, and heat at 100°C for 30min to obtain Fe 3 o 4 / Au composite nanoparticles.

[0063] (2) Preparation of gold magnetic nano-enzyme immunoprobe

[0064] Fe 3 o 4 / Au composite nanoparticles combined with β-hCG to prepare gold magnetic nano-enzyme immunoprobes. The specific method is as follows.

[0065] Activation: weigh 1mg Fe 3 o 4 / Au composite nanoparticles were washed several times with 1 mL of activation buffer (4-morpholineethanesulfonic acid (MES) solution (pH=5.5, 0.01 mol / L)) until the supernatant was clear. Subsequently, continue to dissolve 0.1g carbodiimide (EDC) and 0.1g N-hydroxysulfosuccinamide (NHS) in the abo...

Embodiment 2

[0079] Lateral Flow Immunochromatographic Detection of Antigen NT-proBNP (N-terminal B-type Brain Natriuretic Peptide)

[0080] (1) Fe 3 o 4 / Au Composite Nanoparticles Preparation

[0081] Weigh 130mg of Fe with a particle size of about 150nm 3 o 4 For nanoparticles, add 0.3g citric acid and 3mL chloroauric acid solution with a concentration of 24mmol / L, dilute to 150mL with ultrapure water, and heat at 110°C for 40min to obtain Fe 3 o 4 / Au composite nanoparticles.

[0082] (2) Preparation of gold magnetic nano-enzyme immunoprobe

[0083] Fe by physical adsorption 3 o 4 / Au composite nanoparticles and anti-NT-proBNP monoclonal antibody to prepare gold magnetic nano-enzyme immunoprobe. The specific method is as follows.

[0084] First, use K 2 CO 3 Solution Regulated Fe 3 o 4 The pH value of the / Au composite nanoparticle solution is 8.0; then add 100 μg anti-NT-proBNP monoclonal antibody for coupling; after the coupling is completed, add blocking solution (BS b...

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Abstract

The invention discloses lateral flow immunochromatographic assay test paper based on a gold magnetic nano-enzyme immune probe, as well as a preparation method and an application of the lateral flow immunochromatographic assay test paper, and belongs to the technical field of biological analysis. The preparation method comprises the following steps of: preparing Fe3O4 / Au composite nanoparticles byadopting an in-situ reduction method, then preparing the gold magnetic nano-enzyme immune probe by adopting a physical or chemical method, and finally finishing membrane material treatment and assembly of lateral flow immunochromatographic test paper to obtain the lateral flow immunochromatographic assay test paper based on the gold magnetic nano-enzyme immune probe. In the preparation method, more enzyme catalytic active sites are provided by utilizing a Fe3O4 / Au composite nanoparticle phase, an enzyme catalytic chromogenic reaction rate is effectively improved, and the preparation method hasthe advantages of simple process, low cost and easiness in large-scale production; and compared with the existing lateral flow immunochromatographic test paper, the lateral flow immunochromatographicassay test paper based on the gold magnetic nano-enzyme immune probe has the advantages that the detection sensitivity can be improved by 2-3 orders of magnitudes, the specificity of a detection system is high, and an efficient detection technology is provided for marker analysis with low detection limit.

Description

technical field [0001] The invention belongs to the technical field of biological analysis, and relates to a lateral flow immunochromatographic detection test paper based on a gold magnetic nano-enzyme immunoprobe, a preparation method and an application thereof. Background technique [0002] Lateral flow immunoassay (LFIA) test paper is an ideal instant detection method, which has the advantages of short detection time, simple operation, on-site detection, and strong versatility. It is widely used in medical diagnosis, food Toxicity testing, chemical contamination testing, agriculture, identifying drug abuse and monitoring animal health, etc. However, the traditional colloidal gold-based lateral flow immunochromatographic detection test paper has low detection sensitivity, is prone to false positives, and has poor repeatability, and can only provide qualitative or semi-quantitative detection results, which greatly limits the development of this detection method in the field...

Claims

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Application Information

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IPC IPC(8): G01N33/58G01N33/558G01N33/52
CPCG01N33/583G01N33/587G01N33/558G01N33/52
Inventor 杨冬王丽霞王雪雷蕾高可奕贾彤彤
Owner SHAANXI UNIV OF SCI & TECH
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