Melamine hapten and antigen as well as preparation method and application thereof

A melamine and antibody technology, which is applied to the preparation method of peptides, chemical instruments and methods, animal/human proteins, etc., to achieve the effect of simple sample pretreatment, low cost and simple operation

Active Publication Date: 2010-02-10
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

Defects in the protein content testing methods of the food and feed industry, plus melamine as a white crystalline powder, has no smell and taste, and it is not easy to be found after being mixed with melamine in food and feed. For this reason, melamine is often used by unscrupulous businessmen as Food additives to improve the protein content index in food testing, so melamine is also known as "protein essence"

Method used

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  • Melamine hapten and antigen as well as preparation method and application thereof
  • Melamine hapten and antigen as well as preparation method and application thereof
  • Melamine hapten and antigen as well as preparation method and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0031] Embodiment 1, preparation and identification of melamine hapten and melamine antigen

[0032] Synthesis of melamine hapten and antigen figure 1 shown.

[0033] 1. Synthesis and identification of melamine hapten

[0034] 1. Synthesis of melamine hapten

[0035] 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 ...

Embodiment 2

[0046] Embodiment 2, preparation and identification of melamine hapten and melamine antigen

[0047] 1. Synthesis and identification of melamine hapten

[0048] 1. Synthesis of melamine hapten

[0049] Weigh 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 p-aminophenylbutyric acid (2-chloro-4,6-diamino-1,3,5-triazine and p-aminophenylbutyric acid have a molar ratio of 1:2 ), adding KOH so that the final concentration of KOH is 85%. The water bath was refluxed at 70° C. for 26 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 phase was adjusted to 1 with concentrated hydrochloric acid, the ...

Embodiment 3

[0057] Embodiment 3, preparation and identification of melamine hapten and melamine antigen

[0058] 1. Synthesis and identification of melamine hapten

[0059] 1. Synthesis of melamine hapten

[0060] Weigh 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 p-aminophenylbutyric acid (2-chloro-4,6-diamino-1,3,5-triazine and p-aminophenylbutyric acid have a molar ratio of 1:3 ), adding KOH so that the final concentration of KOH is 85%. The water bath was refluxed at 80° C. for 30 hours, and 0.37 g p-aminobenzenebutyric 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 phase was adjusted to 1 with concentrated hydrochloric acid, the...

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PUM

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Abstract

The invention discloses melamine hapten and antigen as well as a preparation method and application thereof. The method for preparing the compound shown in the formula I comprises the following steps:mixing 2-chloro-4,6-diamino-1,3,5-triazine and p-aminophenylbutyric acid to undergo reflux reaction for 24-30h at a temperature of 70-80 DEG C in the alkaline environment without water and obtainingthe compound shown in the formula I. The molar ratio of the 2-chloro-4,6-diamino-1,3,5-triazine to the p-aminophenylbutyric acid is 1: (1-3), preferably 1:1. The experimental results show that the titer of antiserum obtained by utilizing the antigen of the invention to immunize animals can reach 1:8*10<4>, the lowest limit of detection is 49ng/mL, the hemi-inhibitory concentration is 257ng/mL andthe generated antibody has high specificity, high sensitivity and high accuracy. Therefore, the antigen, antibody and detection method of the invention have extensive application prospects.

Description

technical field [0001] The invention relates to a melamine hapten and antigen, a preparation method and application thereof. Background technique [0002] Melamine, molecular formula C 3 N 6 h 6 , referred to as triamine, is a triazine nitrogen-containing heterocyclic organic compound. Melamine is weakly alkaline and can form melamine salts with hydrochloric acid, sulfuric acid, nitric acid, acetic acid, oxalic acid, etc. Melamine is an important nitrogen-heterocyclic organic chemical raw material. In neutral or slightly alkaline conditions, it can condense with formaldehyde to form various methylol melamines, but in slightly acidic environments (pH5.5-6.5) The derivatives undergo polycondensation reaction to form resin products. Melamine is a basic organic chemical intermediate product with a wide range of uses, and its main use is as a raw material for the production of melamine-formaldehyde resin. The resin has higher hardness than urea-formaldehyde resin, non-flamm...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07D251/70C07K14/765C07K14/795C07K14/77C07K1/113C07K16/44G01N33/566G01N33/531
Inventor 吴小平江海洋王战辉米铁军阮永磊徐飞李娜
Owner 北京维德维康生物技术有限公司
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