Methamidophos half-antigen, artificial antigen and its preparing method

An artificial antigen, the technology of methamidophos, which is applied in the field of immunology, can solve the problems that do not involve the preparation of anti-methamidophos antibody, and do not mention the preparation method of methamidophos artificial antigen.

Inactive Publication Date: 2008-02-20
SOUTH CHINA AGRI UNIV
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  • Claims
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Problems solved by technology

[0008] The patent application No. 03114895.6 published in July 2003 relates to an enzyme-linked immunosorbent assay kit suitable for the analysis of methamidophos residues. It describes the reagents and assay methods in the kit, but does not involve anti-methamidophos antibodies preparation, not to mention the preparation method of methamidophos artificial antigen

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  • Methamidophos half-antigen, artificial antigen and its preparing method
  • Methamidophos half-antigen, artificial antigen and its preparing method

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Embodiment 1

[0034] The preparation method of embodiment 1 methamidophos hapten (n=2)

[0035] 1) Dilute 1 part of O, S-dimethylphosphoryl thiochloride with 5 parts of anhydrous methanol;

[0036] 2) Add 1 part of β-alanine (NH 2 CH 2 CH 2 COOH) was dissolved in 5 parts of anhydrous methanol, and 2 to 3 parts of KOH were added;

[0037] 3) Add the solution of 2) dropwise to the solution of 1) in the stirring state, and continue to stir for 20 minutes after the addition, the product is adjusted to pH 2 with 1N HCl-chloroform, and the chloroform layer is passed over anhydrous MgSO 4 Dry and concentrate under reduced pressure to nearly dryness to obtain a yellow oil. Pass through a silica gel column (silica gel is a product of Qingdao Ocean Chemical Factory Branch), the eluent is methanol:petroleum ether=1:1, the product tubes are combined, and concentrated to dryness under reduced pressure to obtain the product O,S-dimethyl-N- (2-carboxyethyl) phosphorothioate. 1 H NMR: δ2.20(d, 3H, CH...

Embodiment 2

[0038] The preparation method of embodiment 2 methamidophos hapten (n=9)

[0039] 1) Dilute 1 part of O, S-dimethylphosphoryl thiochloride with 15 parts of absolute ethanol;

[0040] 2) 5 parts of 10-aminodecanoic acid (NH 2 (CH 2 ) 9 COOH) was dissolved in 15 parts of absolute ethanol, and 15 parts of NaOH were added;

[0041] 3) Add the solution of 2) dropwise to the solution of 1) in the stirring state, and continue to stir for 30 minutes after the addition, and adjust the pH of the product to 5 with 1N HCl-chloroform, take the chloroform layer and pass through anhydrous MgSO 4 Dry and concentrate under reduced pressure to nearly dryness to obtain a yellow oil. After passing through a silica gel column, the eluent was methanol:petroleum ether=3:1, the product tubes were combined, and concentrated to dryness under reduced pressure to obtain the product O, S-dimethyl-N-(9-carboxynonyl)thiophosphoramide .

Embodiment 3

[0042] Embodiment 3 active ester method prepares methamidophos artificial antigen

[0043] 80 micromole N,N'-dicyclohexylcarbodiimide (DCC) was dissolved in 1mL N,N-dimethylformamide (DMF), and the equivalent amount of the above hapten and N -in a DMF solution of hydroxysuccinimide (NHS), stirred overnight. The reaction supernatant was added dropwise into 5 mL of 10 mg / mL bovine serum albumin (BSA) carbonate buffer solution, stirred slowly overnight, and the product was transferred into a dialysis bag, and dialyzed against phosphate buffer at 4°C for 3 day, freeze-dried, subpackaged, and frozen. The product was made into a 1mg / mL solution with phosphate buffer solution for ultraviolet (200-400nm note: this is the wave path used for scanning, not measured with a single wavelength) spectrum scanning, and 1mg / mL BSA / phosphate buffer solution was used as In contrast, it was found that the ultraviolet spectrum of the product changed significantly compared with BSA, indicating tha...

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Abstract

The invention relates to a methylamine phosphine semi and artificial antigen and its preparing method, using O, S-dimethylthiophosphoryl chloride as the precursor, synthesizing methylamine phosphine structural derivative (semi-agtigen) and a bridge structure is introduced into the derivative on the basis of keeping the basic structure of methylamine phosphine, thus beneficial for the derivative to couple with macro- molecules and able to fully expose lower-molecular weight basic structure of methylamine phosphine so as to avoid methylamine phosphine from being screened by macromolecules to affect recognition of animal organisms.

Description

technical field [0001] The invention relates to a methamidophos hapten, an artificial antigen and a preparation method thereof, belonging to the technical field of immunology. Background technique [0002] Immunoassay is an analysis method based on antigen-antibody specific recognition and binding reaction. It has the advantages of specificity, sensitivity, low requirements for equipment, simple operation, large detection capacity, and low cost. In recent years, it has been used in clinical medical testing, food There are many researches in the fields of analysis and environmental monitoring, and the application prospects are broad. [0003] Pesticide immunoassay is to use the specific binding reaction between pesticides (antigens) and their antibodies to complete the determination of pesticides in a certain system. The preparation of specific antibodies is a necessary prerequisite for the analysis of pesticide residues using immunoassay techniques, which is often achieved ...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C07F9/24C07K2/00C07K1/02
Inventor 孙远明肖治理雷红涛吴青赵肃清
Owner SOUTH CHINA AGRI UNIV
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