Immune antibody for testing residual of polyether-like antibiotic and use thereof

A technology of polyether antibiotics and immune antibodies, applied in immunoglobulin, biological testing, measuring devices, etc., can solve problems such as large consumption of organic solvents, secondary pollution of the environment, and complicated pretreatment processes

Inactive Publication Date: 2005-10-05
JIANGSU ACADEMY OF AGRICULTURAL SCIENCES
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, the analysis method of monensin residues is mainly through extraction and purification, followed by post-column derivatization and high-performance liquid chromatography or liquid chromatography-mass spectrometry. This detection method is expensive, the sample pretreatment process is complicated, and the equipment requirements High, and need to consume a large amount of organic solvents, resulting in secondary pollution to the environment

Method used

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  • Immune antibody for testing residual of polyether-like antibiotic and use thereof
  • Immune antibody for testing residual of polyether-like antibiotic and use thereof
  • Immune antibody for testing residual of polyether-like antibiotic and use thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0019] Embodiment 1, hapten transformation

[0020] The transformation of hapten by hydrochloric acid immersion method is realized as follows: Weigh 500 mg of monensin sodium salt, dissolve it in 1 mL of double distilled water, add 5 mL of hydrochloric acid (HCl) with a concentration of 6 molar, soak for 45-50 min, fully After contact, extract with ether, and remove the ether from the extract in a vacuum oven to precipitate crystals. Prepared into monensin acid. Precipitated crystals were identified by thin-layer chromatography and infrared spectroscopy. figure 2 It is the infrared spectrogram of monensin hapten modified by hydrochloric acid immersion method.

[0021] The transformation of hapten by cation exchange method is realized as follows: Weigh 16.308g of cation exchange resin, stir and wash it with distilled water for 3 times, then add it to the chromatography column, and adjust the flow rate to 1.5mL / min; add 200mL of hydrochloric acid with a concentration of 2 mol...

Embodiment 2

[0024] Embodiment 2. Preparation of artificial immunogen or coated antigen by isobutyl chloroformate method

[0025]1. Artificial immunogen or coated antigen can be prepared in this way: Weigh 192.5mg (0.25mmol) of monensin sodium-succinic anhydride derivative and dissolve it in 2.5mL of dioxane, stir continuously at 10°C; add equimolar 47ul of tributylamine and 35ul of isobutyl chloroformate were continuously stirred at 10°C for 50min to form a mixed anhydride solution; 340mg (0.005mmol) of bovine serum albumin (BSA) was weighed and dissolved in 22mL of 0.1 molar carbonate buffer, Slowly add 22mL of dioxane and keep it at 4°C; slowly drop the mixed acid anhydride solution into the continuously stirring protein solution and react for 5h; dialyze 4 times at 4°C, subpackage, vacuum dry and freeze to prepare artificial immunogen.

[0026] Take 225mg ovalbumin (OVA) to replace the bovine serum albumin (BSA) mentioned above, ovalbumin (OVA) is dissolved in 22mL0.1 molar carbonate b...

Embodiment 3

[0029] Example 3. Preparation of immunogen and coated antigen by water-soluble carbodiimide method

[0030] Artificial immunogen and coated antigen can also be prepared in this way: weigh 160mg (0.25mmol) monensin acid and dissolve it in 2mL of a mixture of 50% (v / v) pyridine and water; weigh carbodiimide (EDC) 160mg Dissolve in 2mL water, add dropwise to monensin acid, stir for 2h while adding; weigh 340mg (0.005mmol) bovine serum albumin (BSA) and dissolve it in 8mL water, slowly add this solution dropwise to the above mixture and keep the pH at about 6.0. After the dropwise addition, place the mixed solution in a dark place at room temperature for 24 hours; place the completed solution in a dialysis bag for dialysis for 4 days. The dialysis solution is PBS solution. Prepared as an artificial immunogen.

[0031] Weigh 225mg ovalbumin (OVA) to replace the bovine serum albumin (BSA) mentioned above, dissolve ovalbumin (OVA) in 8mL water, then slowly add this solution dropwise...

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Abstract

The invention relate to a immunity antibody for detecting polyether antibiotic remains and the application of the antibody. With the monensin sodium salt as half antigen, first reform the half antigen and then couple the reformed half antigen with protein producing artificial immunogen, and obtain multi clone antigen by immunizing the New Zealand rabbit or single clone antigen by immunizing the small mouse, cell fusion, sieving, clone cultivation. The obtained antigen can be used for test the animal source food on polyether antibiotic remains. The advantages of the invention are that the antigen obtained by immunizing animals with the coupling product of reformed monensin sodium salt and protein, has got specificity on polyether antibiotic medical which can be applied on polyether antibiotic remain detecting in animal source food. The detection adopted with this invention is fast and convenience.

Description

Technical field: [0001] The invention relates to an immune antibody for immunological detection of polyether antibiotics and an application thereof, belonging to the technical field of enzyme-linked immunoassay. Background technique: [0002] Polyether antibiotics are widely used in production due to their special physical and chemical properties and their mechanism of action. At present, it has been widely used in poultry, rabbits, etc., and has become the earliest and largest-selling anticoccidial drug in the world; in addition, it is also used as a feed additive in cattle and pig industries to improve cattle and pigs. Pig production performance, promote individual growth. However, the irrational use of such drugs will lead to drug residues in livestock and poultry tissues and livestock products such as meat, eggs, and milk, posing a threat to human health. Studies have shown that monensin can cause and aggravate cardiovascular diseases Or cause cardiovascular dysfunctio...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K16/00G01N33/53G01N33/531G01N33/532
Inventor 刘铁铮王冉魏瑞成张岳梅
Owner JIANGSU ACADEMY OF AGRICULTURAL SCIENCES
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