Method for detecting melamine and enzyme-linked immunosorbent assay kit special for same

An enzyme-linked immunosorbent reagent, melamine technology, applied in the direction of immunoglobulin, chemical instruments and methods, biochemical equipment and methods, etc., can solve the problems that are not suitable for on-site monitoring and screening of a large number of samples, cumbersome processes, complex instruments and equipment, etc.

Inactive Publication Date: 2011-06-08
北京维德维康生物技术有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] The current chemical methods for detecting melamine content include high performance liquid chromatography and liquid chromatography-tandem mass spectrometry. Due to the complicated equipment and cumbersome process, they are not suitable for on-site monitoring and screening of a large number of samples.

Method used

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  • Method for detecting melamine and enzyme-linked immunosorbent assay kit special for same
  • Method for detecting melamine and enzyme-linked immunosorbent assay kit special for same
  • Method for detecting melamine and enzyme-linked immunosorbent assay kit special for same

Examples

Experimental program
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Effect test

Embodiment 1

[0066] Embodiment 1, melamine hapten, melamine antigen and melamine monoclonal antibody and preparation thereof

[0067] 1. Synthesis and identification of melamine hapten

[0068] 1. Synthesis of melamine hapten

[0069] Weigh 0.430 g (3 mmol) of 2-chloro-4,6-diamino-1,3,5-triazine (CAAT) and suspend in Na 2 SO4 In the 10mL dehydrated ethanol extraction of treatment, add 0.5g (3mmol) p-aminophenylbutyric acid (2-chloro-4,6-diamino-1,3,5-triazine and p-aminophenylbutyric acid feed intake The molar ratio is 1:1), and KOH was added to make the final concentration of KOH 85%. The water bath was refluxed at 75° C. for 24 hours, and 0.37 g p-aminophenylbutyric acid (2.0 mmol) and 85% KOH (180 mg, 2.7 mmol) were added at 16 hours and 19 hours, respectively. The product was filtered and distilled under reduced pressure to obtain an oily substance. The product was dissolved in 30 mL of 5% sodium carbonate solution, washed twice with 10 mL of dichloromethane, the pH of the aqueous ...

Embodiment 2

[0103] Embodiment 2, the enzyme-linked immunosorbent assay kit that detects melamine and its preparation

[0104] 1. The enzyme-linked immunosorbent assay kit consists of the following materials:

[0105] 1. Melamine coating agent; as described in Example 1.

[0106] 2. ELISA plate;

[0107] 3. Melamine monoclonal antibody: secreted by the melamine monoclonal antibody hybridoma cell line MAL with the preservation number CGMCC No.3394;

[0108] 4. Melamine standard: the concentration of the standard solution is 0 μg / L, 5 μg / L, 20 μg / L, 100 μg / L, 500 μg / L and 2500 μg / L; the melamine standard is purchased from Sigma Aldrich; CAS number: 108- 78-1; Dilute to the above concentrations with sample diluent;

[0109] 5. Goat anti-mouse secondary antibody labeled with horseradish peroxidase;

[0110] 6. Substrate chromogenic solution: composed of liquid A and liquid B, liquid A is an aqueous solution of 2% carbamide peroxide, liquid B is an aqueous solution of 1% tetramethylbenzidin...

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Abstract

The invention discloses a method for detecting melamine and an enzyme-linked immunosorbent assay (ELISA) kit special for same. The enzyme-linked immunosorbent assay kit comprises melamine monoclonal antibody and melamine hapten. The melamine monoclonal antibody is secreted by a melamine monoclonal antibody hybridoma cell strain MAL with the collection number of CGMCC No.3394. The enzyme-linked immunosorbent assay kit mainly performs qualitative or quantitative detection on the content of melamine in edible products for animals and human (particularly milk products such as milk and milk powder, eggs, feeds and the like) by an indirect competitive ELISA method. The kit and the detection method have low requirement on pretreatment of a sample; the pretreatment process of the sample is simple; and massive samples can be simultaneously detected quickly. The melamine monoclonal antibody with high specificity is adopted, and the detection method is convenient and practicable and has the characteristics of high specificity, high sensitivity, high accuracy and the like.

Description

technical field [0001] The invention relates to a method for detecting melamine and a special ELISA kit. Background technique [0002] Melamine is a triazine nitrogen-containing heterocyclic organic compound, an important nitrogen heterocyclic organic chemical raw material. Pure white monoclinic prism crystal, odorless. Soluble in hot water, slightly soluble in cold water, very slightly soluble in hot ethanol, insoluble in ether, benzene and carbon tetrachloride, soluble in methanol, formaldehyde, acetic acid, hot ethylene glycol, glycerin, pyridine, etc. It is generally stable, but it may decompose and release cyanide at high temperature. [0003] Since the nitrogen content in the molecular formula of melamine is about 66%, and the average nitrogen content of protein is about 16%, it is often used as a food additive by some unscrupulous businessmen to improve the protein content index in food testing, so melamine is Commonly known as "protein essence". Although it is ge...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K16/44C12N5/20G01N33/577
Inventor 江海洋吴小平王战辉史为民徐飞王照鹏潘净茹张兴祥李姮
Owner 北京维德维康生物技术有限公司
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