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Dexamethasone semi-antigen, preparation method and applications thereof

A technology of dexamethasone and hapten, which is applied in the field of hapten and its preparation, can solve problems such as hazards, and achieve the effects of convenient use, low detection cost and accurate detection method

Active Publication Date: 2013-12-04
BEIJING KWINBON BIOTECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, this hormone remains in food and is potentially harmful to humans. Therefore, countries such as the European Union prohibit dexamethasone and dexamethasone as growth-promoting agents for livestock and use them in animal husbandry.

Method used

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  • Dexamethasone semi-antigen, preparation method and applications thereof
  • Dexamethasone semi-antigen, preparation method and applications thereof
  • Dexamethasone semi-antigen, preparation method and applications thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0020] Embodiment 1: Synthesis and identification of dexamethasone hapten (synthetic route such as figure 1 )

[0021] Dissolve 0.80 g dexamethasone in 5 ml DMSO, slowly add 0.5 ml 1,3-propanediamine and 0.5 ml pyridine in 10 ml DMSO dropwise at 60°C, and continue the reaction for 15 hours after the addition is complete. The solvent and unreacted propylenediamine were removed by rotary evaporation, and the propylenediamine monocondensate of dexamethasone was quantitatively obtained.

[0022] Get above-mentioned product and measure through proton nuclear magnetic resonance spectrum, such as figure 2 As shown, the nuclear magnetic spectrum shows that the olefin peak between 6.5-7.0ppm in the spectrum moves to the high field, and the methylene peak increased between 1.0-2.5ppm shows that the hapten is successfully synthesized.

Embodiment 2

[0023] Example 2: Dexamethasone Antigen

[0024] The dexamethasone hapten is coupled with the carrier protein to obtain the dexamethasone antigen.

[0025] 1. Immunogen preparation - synthesis of dexamethasone hapten-bovine serum albumin (BSA) conjugate

[0026] Dissolve 50 mg of EDC in 2 ml of water to obtain solution I; dissolve 11 mg of dexamethasone hapten in 1.0 ml of DMF to obtain solution II; dissolve 60 mg of BSA in 3 ml of 0.01mol / L PBS (pH=8.0) to obtain solution III ; Mix solution II and solution III, and add solution I dropwise under magnetic stirring; stir and react at room temperature for 24 hours. Dialyzed with triple distilled water for 48 hours to obtain the immunogen.

[0027] 2. Coating source preparation - synthesis of dexamethasone hapten-ovalbumin (OVA) conjugate

[0028] Dissolve 15 mg of dexamethasone hapten in 1.0 ml of dimethylformamide (DMF) to obtain solution IV; take 10 μl of 50% glutaraldehyde (GA) and add it to solution IV, stir and react at r...

Embodiment 3

[0031] Embodiment 3: Dexamethasone monoclonal antibody

[0032] 1. Preparation of dexamethasone monoclonal antibody

[0033] Animal immunization: Inject the immunogen into the body of Balb / c mice with an immunization dose of 150 μg / mouse to make them produce polyclonal antibodies.

[0034] Cell fusion and cloning: After the measurement result of mouse serum was higher, the splenocytes were taken and fused with SP2 / 0 myeloma cells at a ratio of 8:1, and the cell supernatant was measured by indirect competitive ELISA, and the positive wells were screened. Use the limited dilution method to clone the positive wells until hybridoma cell lines secreting monoclonal antibodies are obtained, and it is found that the titer of one of the hybridoma cell lines is significantly higher than that of the other hybridoma cell lines, and the dexamethasone monoclonal antibody hybridoma cell line The strain was named D-4-2, and the cell line was preserved on May 3, 2012 in the General Microbiolo...

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Abstract

The invention discloses a semi-antigen, a preparation method and applications thereof, and specifically relates to a dexamethasone semi-antigen. The invention also discloses a preparation method of the semi-antigen and applications thereof. The rapid detection kit product based on the dexamethasone semi-antigen has the advantages of user-friendliness, low detection cost, high efficiency, accuracy and rapidness, is capable of detecting a large amount of samples at the same time, and is suitable for on-site supervision of dexamethasone residual in the animal source food and screening of a large amount of samples.

Description

technical field [0001] The invention relates to a hapten and its preparation method and application, in particular to a dexamethasone hapten and its preparation method and application. Background technique [0002] Dexamethasone (Dex) is a derivative of the glucocorticoid cortisone, also called dexamethasone, and it is also the most widely used potent corticosteroid in clinical medicine. Cortisone hormones are usually used to treat allergic and severe infectious diseases, and have anti-inflammatory and anti-allergic effects. These hormones still have the functions of stimulating appetite, increasing liver glycogen synthesis, and raising blood sugar. However, this hormone remains in food and is potentially harmful to humans. Therefore, countries such as the European Union prohibit the use of dexamethasone and dexamethasone as growth-promoting agents for livestock in animal husbandry. [0003] The maximum residue limit of dexamethasone stipulated by the European Union in the...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07J41/00C07K14/435C07K14/765C07K14/77C07K14/795C07K16/26G01N33/74
Inventor 何方洋崔海峰杨昌松张荃郝士元蒲小容冯静罗贵昆
Owner BEIJING KWINBON BIOTECH
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