Chemiluminescence immune detection reagent kit for detecting salbutamol

A technique of chemiluminescence immunity and albuterol, which is applied in the field of immunological detection, can solve problems such as prolonging detection time, and achieve the effects of increasing sensitivity, shortening detection time, and reducing incubation time.

Inactive Publication Date: 2009-04-22
SHANGHAI JIAO TONG UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The disadvantage of the method used by Xu Chuanlai et al. is that the primary antibody and the secondary antibody need to be used at the same time (the secondary antibody is an enzyme-labeled antibody). After the addition of the two antibodies, incubation is required, which prolongs the detection time.

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0049] 1) Preparation of each component of the kit

[0050] Preparation of salbutamol hapten: acidify salbutamol, react with sodium nitrite in a low-temperature environment without light at 4°C, and generate intermediates containing diazonium cations. The diazotized albuterol was used as a hapten for subsequent synthesis of immune antigens and coating antigens.

[0051] Synthesis of bovine serum albumin-salbutamol immune antigen: the immune antigen was obtained by coupling salbutamol and bovine serum albumin (BSA) by diazotization method.

[0052] Synthesis of human serum albumin-salbutamol-coated antigen: the immune antigen was obtained by coupling salbutamol and human serum albumin (HSA) by diazotization.

[0053] Preparation of albuterol-peroxidase-labeled antibody: inject the above-mentioned immune antigen into Balb / C mice, and make it produce antibody serum after several booster immunizations. Take out the splenocytes of the immunized mice, add them to the centrifuge tu...

Embodiment 2

[0081] The chemiluminescent immunoassay kit for detecting salbutamol includes the following components:

[0082] (1) 48-well opaque white ELISA plates (6 strips x 8 wells) are coated with salbutamol-mouse serum albumin cross-linked complex, and are vacuum-sealed with aluminum film.

[0083] (2) 5 bottles of albuterol standard solution, the concentrations are respectively:

[0084] 0μg / L, 0.04μg / L, 0.36μg / L, 3.24μg / L, 29.16μg / L

[0085] (3) Salbutamol-horseradish peroxidase labeled antibody solution.

[0086] (4) Luminescence substrate A solution (luminol and enhancer), and luminescence substrate B solution (urea hydrogen peroxide).

[0087] (5) 50 times concentrated buffer. Diluted 50 times before use to become a working concentration buffer, the diluted working concentration buffer is 0.05mol / L Tris-HCl buffer, pH6.9, containing 0.1% (v / v) Tween-20.

Embodiment 3

[0089] The chemiluminescent immunoassay kit for detecting salbutamol includes the following components:

[0090] (1) 96-well opaque white ELISA plate (12 strips x 8 wells) is coated with albuterol-egg albumin cross-linked complex and vacuum-sealed with aluminum film.

[0091] (2) 6 bottles of salbutamol standard solution, the concentrations are respectively:

[0092] 0μg / L, 0.05μg / L, 0.25μg / L, 1.25μg / L, 6.25μg / L, 31.25μg / L.

[0093] (3) Salbutamol-horseradish peroxidase labeled antibody solution.

[0094] (4) Luminescence substrate A solution (luminol and enhancer), and luminescence substrate B solution (urea hydrogen peroxide).

[0095] (5) 10 times concentrated buffer. Diluted 10 times before use to become a working concentration buffer, the diluted working concentration buffer is 0.05mol / L glycine-HCl buffer, pH6.9, containing 0.02% (v / v) Tween-20.

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PUM

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Abstract

The invention provides a chemiluminescent immunoassay kit for detecting salbutamol, and the kit belongs to the field of immunoassay. The kit is composed of a non-transparent white enzyme-labeled plate which is coated by a salbutamol-carrier protein cross-linked complex, a salbutamol standard product, a salbutamol-peroxidase-labeled antibody, luminescent substrate solution and concentrated buffer solution. The salbutamol-carrier protein cross-linked complex is obtained by coupling the salbutamol and a carrier protein through the mixed-anhydride method or the direct protein activation method, and the concentrated buffer solution contains Tween-20 buffer solution. The kit has the advantages of rapidness, simpleness, high sensitivity, low detection cost, good repeatability, high throughput, and the like, and the kit can be used for detecting the residual salbutamol in animal urine, blood, tissues, visceral organs and other samples.

Description

technical field [0001] The invention relates to a chemiluminescent immunoassay kit for detecting albuterol, which is used for detecting the content or residual amount of albuterol in animal-derived food such as animal tissue, viscera, blood, urine, feed, and feed raw materials. It belongs to the field of immunological detection. Background technique [0002] Salbutamol (Salbutamol, SAL) belongs to β-agonists. β-agonist is a kind of nutrient redistribution agent, which is a kind of phenylethanolamine derivatives similar in structure and function to adrenaline and norepinephrine. It can improve the ratio of meat to fat in fatty animals and reduce ketone bodies Fat content, increase lean meat rate, and can accelerate animal growth, so it is added to animal feed. Common β2-agonists include clenbuterol, ractopamine, and salbutamol. With the increasing supervision of clenbuterol (commonly known as "clenbuterol") in my country, the use of clenbuterol has gradually decreased, and...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/577G01N33/543G01N21/76
Inventor 陈峰梁晓翠卢庆鋆徐晓婴卢永红
Owner SHANGHAI JIAO TONG UNIV
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