Method for preparing aflatoxin G1 artificial antigen

An aflatoxin and artificial antigen technology, which is applied to the preparation methods of peptides, chemical instruments and methods, animal/human proteins, etc., can solve the problems of lack of aflatoxin G1 artificial antigen preparation method, etc., and is suitable for large-scale production. And the effect of popularization and application, good stability, simple and feasible preparation process

Inactive Publication Date: 2008-09-24
INST OF OIL CROPS RES CHINESE ACAD OF AGRI SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, artificial antigens such as aflatoxin B1 have been successfully reported abroad, but the principle of artificial antigens for aflatoxin G1 has not been reported so far. The fundamental reason is the lack of methods for preparing artificial antigens of aflatoxin G1

Method used

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  • Method for preparing aflatoxin G1 artificial antigen
  • Method for preparing aflatoxin G1 artificial antigen
  • Method for preparing aflatoxin G1 artificial antigen

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0015] 1) get commercially available purity (by weight) and be that 9.86 milligrams of m-chloroperoxybenzoic acid of 85% are dissolved in 1 milliliter of dichloromethane, and use 0.1 moles of 1 milliliter of every liter of pH7.0 phosphate buffer in Wash 4 times in the separating funnel; Weigh 2 mg of aflatoxin G1 and dissolve it in 1 ml of dichloromethane, and add 1 ml of 0.1 moles of pH7.2 per liter of phosphate buffer; the above m-chloroperoxybenzene Mix formic acid solution and aflatoxin G1 solution, and stir magnetically at 22°C for 80 minutes; leave the final reaction solution at room temperature, discard the upper aqueous phase after layering, and keep the organic phase, which is the aflatoxin Solution of G1 epoxide in dichloromethane.

[0016] 2) 2 milliliters of the organic phase retained in the above steps 1) was added to 4.0 milliliters of phosphate buffered saline solution containing 13.6 milligrams of bovine serum albumin at pH 7.2, and the formed two-phase system ...

Embodiment 2

[0019] 1) get commercially available purity (by weight) and be that 9.39 milligrams of m-chloroperoxybenzoic acid of 85% are dissolved in 1 milliliter of dichloromethane, and use equal volume of 0.1 mole per liter of pH7.0 phosphate buffer in Wash 3 times in the separating funnel; Take 2mg aflatoxin G1 and dissolve in 1 milliliter of dichloromethane, and add 0.1 moles of 1 milliliter of every liter of pH7.2 phosphate buffer; The solution was mixed with the aflatoxin G1 solution, and stirred magnetically at 22°C for 100 minutes; the final reaction solution was allowed to stand at room temperature, and the upper aqueous phase was discarded after layering, and the organic phase was kept, which was the aflatoxin G1 Dichloromethane solution of epoxide.

[0020] 2) 2 milliliters of the organic phase retained in the above step 1) was added to 6.0 milliliters of phosphate buffered saline solution containing 13.1 milligrams of ovalbumin at pH 7.2, and the formed two-phase system was ma...

Embodiment 3

[0023] 1) get commercially available purity (by weight) and be that 8.62 milligrams of m-chloroperoxybenzoic acid of 85% are dissolved in 1 milliliter of dichloromethane, and use 0.1 moles of 1 milliliter of every liter of pH7.0 phosphate buffer in Wash 4 times in the separating funnel; Weigh 2 mg of aflatoxin G1 and dissolve it in 1 ml of dichloromethane, and add 1 ml of 0.1 moles of pH7.2 per liter of phosphate buffer; the above m-chloroperoxybenzene Mix formic acid solution and aflatoxin G1 solution, and stir magnetically at 22°C for 80 minutes; leave the final reaction solution at room temperature, discard the upper aqueous phase after layering, and keep the organic phase, which is the aflatoxin Solution of G1 epoxide in dichloromethane.

[0024] 2) 2 milliliters of the organic phase retained in the above step 1) was finally added to 4.0 milliliters of phosphate buffered saline solution containing 12.5 milligrams of bovine serum albumin at a pH of 7.2, and the formed two-p...

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Abstract

The present invention relates to a method for preparing artificial antigens by the coupling of aflatoxin G1 and protein carrier macromolecules. Meta-Chloroperoxybenzoic acid is used to oxidize the double bonds of aflatoxin G1 molecular difuran rings, so that highly active aflatoxin G1 epoxide is formed, then, under the indoor temperature, the aflatoxin G1 epoxide is linked with the amidogen of carrier protein in a biphasic system, and finally, the aflatoxin G1 artificial antigens are prepared after dialysis, refrigeration and drying. The method has the following advantages: (1) Aflatoxin G1 artificial antigens can be effectively prepared, so that antibodies which can identify aflatoxin G1 are produced, which is significant for the immune analysis technology applied to test aflatoxin G1; (2) The whole preparation process is simple and feasible, does not need special apparatuses, has low cost and is suitable for mass production and promotion; (3) The aflatoxin G1 artificial antigens prepared by the method have good stability.

Description

technical field [0001] The invention relates to a method for preparing an artificial antigen by coupling aflatoxin G1 with a protein carrier macromolecule. Background technique [0002] Mycotoxins are secondary metabolites secreted by toxin-producing fungi, and are natural toxic compounds that can cause various damages to humans and animals. Among the mycotoxins that have been discovered, aflatoxin (AFT) is the most toxic mycotoxin, and its toxicity, carcinogenicity and pollution frequency all rank first in biological toxins. Aflatoxin G1 is one of the main types of aflatoxins that have been discovered. After it pollutes food and feed, it directly or indirectly enters the human food chain, threatening human health and life safety. The degree of harm is proportional to the intake of aflatoxins. relevant. Aflatoxins widely exist in rice, corn, peanuts, sesame, soybeans, rapeseed and other agricultural products and fish and other foods, and there are many links in which food ...

Claims

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Application Information

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IPC IPC(8): C07K1/10C07K14/765C07K14/77C07K14/795
Inventor 李培武张文张奇陈小媚丁小霞谢立华姜俊
Owner INST OF OIL CROPS RES CHINESE ACAD OF AGRI SCI
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