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Anti-hydrocortisone specific monoclonal antibody hybridoma cell strain YH7 and application thereof

A hybridoma cell line, hydrocortisone technology, applied in specific peptides, chemical instruments and methods, biochemical equipment and methods, etc., can solve the problems of high environmental requirements, radioisotope pollution, etc., and achieve good detection sensitivity and Affinity, synthesis steps simplified and effective effect

Active Publication Date: 2016-08-31
JIANGNAN UNIV
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, these methods have certain defects. For example, chromatography and mass spectrometry require specialized technicians and expensive instruments, and have relatively high environmental requirements; although radioimmunoassays are more accurate and stable, they have radioactive The problem of isotope contamination, which cannot be solved in non-professional laboratories

Method used

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  • Anti-hydrocortisone specific monoclonal antibody hybridoma cell strain YH7 and application thereof
  • Anti-hydrocortisone specific monoclonal antibody hybridoma cell strain YH7 and application thereof
  • Anti-hydrocortisone specific monoclonal antibody hybridoma cell strain YH7 and application thereof

Examples

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Embodiment 1

[0020] Example 1: Preparation of anti-hydrocortisone-specific monoclonal antibody hybridoma cell line YH7

[0021] (1) Synthesis of complete antigen: Weigh 0.1g of hydrocortisone and 0.3g of succinic anhydride into a 50mL round bottom flask, add 5mL of anhydrous pyridine, heat and stir the mixture at 60°C for 12h, transfer the reactant to a 10 In ice water (4°C) of % HCl (V / V), a white precipitate precipitated, centrifuged to remove pyridine and excess succinic anhydride, washed with deionized water several times, centrifuged, dried in a drying oven, stored after LC-MS identification , as a hapten;

[0022] Take 2mg of the above hapten, dissolve it in 3mL of DMF (N,N-dimethylformamide), then add 2.5mg of EDC (1-(3-dimethylaminopropyl)-3-ethylcarbodiimide Hydrochloride) and 1.5mg NHS (N-hydroxysuccinimide), stirred at room temperature, activated for 4h; another 7.5mg KLH (keyhole limpet hemocyanin) was dissolved in 2mL, 0.05M, pH9.6 CB (carbon salt buffer solution) solution, ...

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Abstract

The invention discloses an anti-hydrocortisone specific monoclonal antibody hybridoma cell strain YH7 and an application thereof, and belongs to the field of food safety immunodetection. The anti-hydrocortisone specific monoclonal antibody hybridoma cell strain YH7 is preserved at China General Microbiological Culture Collection Center; the preservation number is CGMCC No.12022; the hybridoma cell strain CGMCC No.12022 can secrete and generate the monoclonal antibody; the monoclonal antibody has relatively good specificity and relatively high sensitivity on hydrocortisone, has 50% inhibition concentration IC50 of 0.1mug / L on the hydrocortisone and can be used for preparing the hydrocortisonean immunoassay kit and a colloidal gold test strip; and a powerful detection method and means are provided for specific detection of the hydrocortisone in food safety and fast and healthy development of the animal husbandry.

Description

technical field [0001] The invention relates to an anti-hydrocortisone-specific monoclonal antibody hybridoma cell line YH7 and its application, belonging to the field of food safety immune detection. Background technique [0002] Hydrocortisone (HDS) is an important adrenocortical hormone, which can regulate the biosynthesis and metabolism of sugar, fat and protein, and also has various pharmacological effects such as anti-inflammatory, immunosuppressive, anti-drug and anti-shock . Used in feed can promote animal growth, enhance animal immunity and increase animal weight, etc. It can also be used to treat dairy cow mastitis and increase milk production in dairy cow feeding. Its improper use may lead to residual hydrocortisone pollution in aquatic products, meat, eggs and milk. The European Union, the United States, and Japan have all stipulated that the maximum residue limit in milk is 10 μg / kg. [0003] The reported methods for the determination of hydrocortisone mainly...

Claims

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Application Information

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IPC IPC(8): C12N5/20C07K16/44G01N33/74G01N33/577
CPCC07K16/44
Inventor 刘丽强王忠兴胥传来匡华徐丽广马伟吴晓玲宋珊珊胡拥明
Owner JIANGNAN UNIV
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