Ethyl maltol monoclonal antibody hybridoma cell strain and application thereof

A technology of hybridoma cell line and ethyl maltol, which is applied to the preparation method of animal/human protein and peptide, serum albumin, etc., can solve the problems of complexity, expensive equipment, and high professional requirements of operators, and achieve good results The effect of specificity and good detection sensitivity

Active Publication Date: 2021-03-09
JIANGNAN UNIV
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Problems solved by technology

These methods have high sensitivity and stability, but the equipment is expensive, requires high professionalism of operators, and the detection process is cumbersome and complicated, which cannot meet the detection requirements of on-site analysis of large quantities of samples.

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  • Ethyl maltol monoclonal antibody hybridoma cell strain and application thereof

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Embodiment 1

[0029] Embodiment 1 Preparation of hybridoma cell line ABC01

[0030] (1) Synthesis of hapten EM-1: Dissolve 200 mg of ethyl maltol in 5 mL of acetone, then add 600 mg of ethyl 4-bromobutyrate, 800 mg of potassium carbonate and 14.2 mg of potassium iodide, and reflux the solution in a boiling water bath for 24 hours. The organic solvent was removed with a vacuum rotary evaporator to obtain a pale yellow precipitate.

[0031]Dissolve the above precipitate with 0.5M 20% methanol-NaOH aqueous solution, then stir in a water bath at 80°C for 1 h, hydrolyze the ethyl 4-bromobutyrate attached to ethyl maltol to 4-bromobutyric acid, and restore the solution to After room temperature, use 0.5M HCl to adjust to neutrality, and then extract three times with 15mL ethyl acetate. After collecting the organic phase, first use anhydrous Na 2 SO 4 Dry, and finally remove the organic solvent with a vacuum rotary evaporator to obtain the ethyl maltol hapten EM-1.

[0032] (2) Preparation of c...

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Abstract

The invention discloses an ethyl maltol monoclonal antibody hybridoma cell strain and application thereof, and belongs to the field of food safety immunodetection. The disclosed ethyl maltol monoclonal antibody hybridoma cell strain ABC01 is preserved in China General Microbiological Culture Collection Center (CGMCC) and is classified and named as a monoclonal cell strain on November 28, 2019, andthe preservation number is CGMCC No.19178. A complete antigen of ethyl maltol and an equivalent Freund's adjuvant are mixed and emulsified, and a BALB / c mouse is immunized through neck and back subcutaneous multipoint injection. Cells are screened with an indirect competitive enzyme-linked immunosorbent assay method and subcloned for three times, and finally, the monoclonal antibody hybridoma cell strain is obtained. The monoclonal antibody secreted by the cell strain has better specificity and detection sensitivity to ethyl maltol, can realize detection of the residual amount of ethyl maltolin grease, milk and other foods, provides a raw material for immunodetection of ethyl maltol residues in foods, and has practical application values.

Description

technical field [0001] The invention relates to an ethyl maltol monoclonal antibody hybridoma cell line ABC01 and an application thereof, belonging to the field of food safety immunological detection. Background technique [0002] Ethyl maltol (Ethyl maltol, EM) is a flavor and fragrance that can be artificially synthesized. It is widely used in sweetening food, beverages, cigarettes, daily cosmetics and other fields because of its low-volume, high-efficiency, safety and non-toxic characteristics. , enhances, preserves and masks odors. Although ethyl maltol is an ideal edible flavor additive allowed by the regulations of our country, the country also has clear regulations on the scope of its use, among which no flavors and fragrances can be added to vegetable oils, pasteurized milk, sterilized milk, and fermented milk In 2020, the "National Food Safety Supervision and Sampling Implementation Rules" also included ethyl maltol in the edible vegetable oil testing items. Consi...

Claims

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Application Information

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IPC IPC(8): C12N5/20C07K16/44C07K14/795C07K14/765C07K1/02C07K1/107C07K1/34G01N33/577G01N33/53C07D309/40
CPCC07K16/44C07K14/795C07K14/765G01N33/577G01N33/5308C07D309/40
Inventor 胥传来雷咸禄匡华徐丽广马伟刘丽强吴晓玲宋珊珊胡拥明
Owner JIANGNAN UNIV
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