Histamine immunoassay method based on characteristics of platinum-gold bimetallic nano-particle peroxidase

A technology of bimetallic nanoparticles and gold nanoparticles, which is applied in the direction of analytical materials, measuring devices, instruments, etc., can solve the problem of detecting histamine with Pt-Au bimetallic nanoparticles, and achieve easy large-scale preparation and huge application Prospect, effect of simple preparation method

Inactive Publication Date: 2018-08-03
SOUTH CHINA AGRI UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0006] At present, there is no report on the use of Pt-Au bimetallic nanoparticles as a signal probe for the detection of histamine

Method used

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  • Histamine immunoassay method based on characteristics of platinum-gold bimetallic nano-particle peroxidase
  • Histamine immunoassay method based on characteristics of platinum-gold bimetallic nano-particle peroxidase
  • Histamine immunoassay method based on characteristics of platinum-gold bimetallic nano-particle peroxidase

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0044] 1. Preparation of platinum-gold bimetallic nanoparticles

[0045] 0.9 mL of K 2 PtCl 4 (20 mM), 0.1 mL HAuCl 4 (20 mM) mixed, add 10 mg Pluronic F127, ultrasonic to dissolve, then add 1.0 mL of ascorbic acid (100 mM) as reducing agent, the mixed solution was ultrasonically reacted in a water bath at 30ºC for 5 hours, and the product was separated by centrifugation at 10000r / min for 20min. Wash with ethanol and water repeatedly 5 times to remove residual PluronicF127, and reconstitute with 0.9 mL of water for later use.

[0046] From figure 1 It can be seen that the prepared platinum nanoparticles are porous nanostructures with a particle size of about 50 nm, and the surface is composed of Pt nanoparticles with a uniform distribution of about 5 nm.

[0047] 2. Preparation of platinum-gold labeled antibody nanoprobe complex

[0048] Dilute the platinum-gold nanoparticle solution prepared above to 20 mM, take 1.0 mL and adjust the pH value to 8-9 with 0.1M potassium ...

Embodiment 2

[0058] 1. Preparation of platinum-gold bimetallic nanoparticles

[0059] Same as embodiment 1;

[0060] 2. Preparation of platinum-gold labeled antibody nanoprobe complex

[0061] Basically the same as Example 1, the only difference is that the platinum-gold nanoparticle solution was diluted to 5 mM, and 20 μL of histamine monoclonal antibody (final concentration 5.35 mg / mL) was added.

[0062] 3. Establishment of working curve

[0063] Same as Example 1.

Embodiment 3

[0065] 1. Preparation of platinum-gold bimetallic nanoparticles

[0066] Same as embodiment 1;

[0067] 2. Preparation of platinum-gold labeled antibody nanoprobe complex

[0068] Basically the same as in Example 1, the only difference is that the platinum-gold nanoparticle solution was diluted to 30 mM, and 20 μL of histamine monoclonal antibody (final concentration 5.35 mg / mL) was added.

[0069] 3. Establishment of working curve

[0070] Same as Example 1.

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Abstract

The invention discloses a histamine immunoassay method based on the characteristics of platinum-gold bimetallic nano-particle peroxidase. The method comprises the following steps: S1, labeling a histamine monoclonal antibody with a platinum-gold nano-particle to prepare a platinum-gold standard antibody nano-probe; S2, adding a sample to be tested and the nano-probe obtained in step S1 to an ELISAplate coated with a histamine antigen, and forming an antigen and platinum-gold standard antibody immune compound in a competitive immunoassay mode; S3, adding a coloring substrate solution containing 3,3',5,5'-tetramethylbenzidine and hydrogen peroxide to the ELISA plate, performing color development, and terminating the color development with sulfuric acid; and S4, detecting the absorbance at 450 nm by a the absorption of 450 nm was measured by an enzyme-linked immunometric meter to achieve the quantitative detection of histamine in the sample. The method has the advantages of easiness in operation, high sensitivity, good stability and high sample throughput, is suitable for rapid high-throughput screening of the histamine in foods, and has a great application prospect.

Description

technical field [0001] The invention belongs to the technical field of food processing, and more specifically discloses a histamine immunoassay method based on the peroxidase-like properties of platinum-gold bimetallic nanoparticles. Background technique [0002] Histamine is formed by the decarboxylation reaction of free histidine under the catalysis of histidine decarboxylase. It is a toxic biogenic amine that widely exists in various organisms and foods, mainly aquatic products. Especially pelagic fish with green skin and red meat and their processed products. Studies have shown that excessive intake of histamine can cause a series of allergic reactions such as headache, nausea, nausea and vomiting, diarrhea, heart palpitations, itchy rash, and changes in blood pressure. Ingestion of 8-40 mg histamine will produce mild poisoning symptoms, more than 40 mg will produce moderate poisoning symptoms, and more than 100 mg will produce severe poisoning symptoms. The US FDA has...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/577
CPCG01N33/577
Inventor 罗林徐振林杨金易孙远明雷红涛沈玉栋王弘
Owner SOUTH CHINA AGRI UNIV
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