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Synthesis method of artificial antigen suitable for erythrosine pigment

A technology of artificial antigen and synthetic method, which is applied in the direction of chemical instruments and methods, hybrid peptides, instruments, etc., to achieve the effect of simple synthesis steps

Inactive Publication Date: 2011-07-13
JIANGNAN UNIV
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  • Claims
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Problems solved by technology

So far, there is no report on the immunological detection method for erythrosine structural residues at home and abroad. For this purpose, a complete antigen for the detection of erythrosine artificial synthetic pigments was designed and synthesized using erythrosine intermediate fluorescein as a hapten.

Method used

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  • Synthesis method of artificial antigen suitable for erythrosine pigment
  • Synthesis method of artificial antigen suitable for erythrosine pigment
  • Synthesis method of artificial antigen suitable for erythrosine pigment

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Embodiment 1

[0015] (1) Preparation of artificial antigen

[0016] Take 200mg (0.60187mmol) fluorescein FLS, 1.140g (10mmol) glutaric anhydride, and 112mg (1 mmol) lutidine DMAP into a round bottom flask, then add 9mL absolute ethanol and 1mL anhydrous pyridine, and finally Put it in a 75°C water bath for 24h. The product solution was purified by vacuum rotary evaporation, and then added to DMF solution to dissolve for the next step for reaction. Take 0.5 mL of N,N-dimethylformamide DMF solution containing 20 mg (0.06019 mmol) of fluorescein FLS derivatives into a brown vial, and then add 15.52 mg (0.06807 mmol) of N-hydroxysulfosuccinimide Sulf- NHS and 17.32 mg (0.09033 mmol) carbodiimide EDC were reacted at room temperature for 8 hours to form A solution. Dissolve 65mg (0.001mmol) of bovine serum albumin BSA into 0.01M, pH7.4 phosphate buffered saline PBS, which is liquid B. Then slowly add solution A to solution B dropwise, react at room temperature for 6 hours, centrifuge to remove...

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Abstract

A synthesis method of an artificial antigen suitable for erythrosine pigment belongs to the technical field of biochemical engineering. The synthesis method is as follows: fluorescein (FLS) is used as hapten, and the active ester method is adopted to couple the hapten and the carrier protein bovine serum albumin (BSA) to obtain the artificial antigen FLS-BSA conjugate of erythrosine. Polyacrylamide gel electrophoresis is adopted to detect the coupling ratio of the conjugate. By adopting the method, the FLS-BSA conjugate can be synthesized successfully, namely the artificial antigen suitable for erythrosine pigment can be obtained. The synthesis step is concise and effective; and the method can be completely used in immunoassay, provide the necessary artificial antigen for the future research of people and meet the need of the domestic research on the artificial antigen.

Description

technical field [0001] The invention relates to a method for synthesizing an artificial antigen suitable for erythrosin, which belongs to the technical field of biochemical engineering. Background technique [0002] Erythrosine, the English name is: Erythrosine, ETS, its chemical name is: 2-(2,4,5,7-tetraiodo-6-hydroxy-3-oxanthene-9-yl) benzoic acid disodium salt , also known as tetraiodofluorescein B, CAS No. 568-63-8, molecular formula is C 20 h 6 I 4 Na 2 o 5 . Erythrosine is a kind of synthetic pigment with bright color, which is often used in food processing products and beverages to improve its sensory properties. Food colorants can be divided into two categories: natural pigments and synthetic pigments. Synthetic pigments are widely used in the food processing industry because of their low cost, diverse tones, bright colors and strong tinting strength. Because synthetic pigments use chemical products such as benzene, toluene, and naphthalene as raw materials, t...

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Application Information

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IPC IPC(8): C07K19/00G01N33/533
Inventor 匡华胥传来李灼坤邢常瑞宋珊珊刘丽强王利兵
Owner JIANGNAN UNIV
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