A zilpaterol monoclonal antibody hybridoma cell line and its application

A hybridoma cell line and monoclonal antibody technology, which is applied to specific peptides, instruments, physical/chemical process catalysts, etc., can solve the problems of high detection limit, cumbersome detection methods, and low sample purity requirements, and achieve good detection sensitivity. Effect

Active Publication Date: 2022-04-12
JIANGNAN UNIV
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Problems solved by technology

High-performance liquid chromatography is used for detection. The detection method is cumbersome and complicated, and the detection limit is too high. In order to protect the interests of consumers, it is necessary to establish an efficient and rapid detection method for ZIL. ) is simple in pretreatment, low in cost, can realize rapid detection of a large number of samples, and does not require high purity of samples during detection

Method used

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  • A zilpaterol monoclonal antibody hybridoma cell line and its application
  • A zilpaterol monoclonal antibody hybridoma cell line and its application

Examples

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Embodiment 1

[0026] Example 1 Preparation of hybridoma cell line SMB1C2

[0027] (1) Preparation of the complete antigen re-ZIL-EDC-BSA: Weigh 2.65 mg of zilpaterol derivative (the molar ratio of re-ZIL to bovine serum albumin (BSA) is 100:1), dissolve in 300 μL N, In N-dimethylformamide (DMF), stir the reaction for 10 minutes. After fully dissolving, add 2.3 mg of N-hydroxysuccinimide (NHS) and 1-(3-dimethylaminopropyl)-3-ethane Carbodiimide hydrochloride (EDC) 3.5 mg, activated at room temperature for 4-6 hours, as A solution. Take 6mg of BSA, dissolve it with 2mL 0.01M carbonate buffer solution (CB, pH=9.0) (referred to as solution B), then slowly add solution A to solution B drop by drop, and perform coupling reaction at room temperature overnight; then use 0.01 M PBS solution was dialyzed to remove the unreacted small molecule hapten to obtain the complete antigen re-ZIL-EDC-BSA, which was identified by UV absorption scanning method;

[0028](2) Animal immunization: After mixing and...

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Abstract

A zilpaterol monoclonal antibody hybridoma cell line and application thereof belong to the technical field of food safety immunoassay. Zilpaterol monoclonal antibody hybridoma cell line SMB1C2 has been deposited in the General Microbiology Center CGMCC of the Chinese Microbiological Culture Collection Management Committee, classified and named as a monoclonal cell strain, and the date of preservation is November 28, 2019, and the preservation number is CGMCC No. 19170. The complete antigen of zilpaterol was mixed and emulsified with the same amount of Freund's adjuvant, and BALB / c mice were immunized by multi-point subcutaneous injection on the back of the neck, and finally a monoclonal antibody hybridoma cell line was obtained. The monoclonal antibody secreted by this cell line has good specificity and detection sensitivity to zilpaterol (IC 50 The value is 0.5ng / mL), which can realize the detection of zilpaterol residues in milk, muscle tissue and urine, and provides raw materials for the immunological detection of zilpaterol residues in food, which has practical application value.

Description

technical field [0001] The invention relates to a zilpaterol monoclonal antibody hybridoma cell line and application thereof, and belongs to the technical field of food safety immunoassay. Background technique [0002] Zipaterol (Zilpaterol hydrochloride, ZIL) is a stimulant drug, β-adrenoceptor agonist. This class of drugs refers to a class of drugs that can stimulate bronchial smooth muscle β-adrenergic receptors. β-adrenergic receptor agonist drugs can relax bronchial smooth muscle and bronchiectasis. It can be used to treat bronchial asthma and chronic asthmatic bronchitis. Commonly used salbutamol, terbutaline and so on. Including non-selective β-adrenoceptor agonists such as adrenaline, ephedrine and isoprenol and selective β2-adrenoceptor agonists such as salbutamol, terbutaline, etc. They mainly stimulate the β2 receptors in the respiratory tract, activate adenylyl cyclase, increase the content of cyclic adenylphosphine (cAMP) in cells, and free Ca 2+ Reduce, th...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N5/20C07K16/44C07K14/765C07D487/06B01J23/42B01J23/44G01N33/577
CPCC07K16/44C07K14/765C07D487/06B01J23/42B01J23/44G01N33/94G01N33/577
Inventor 胥传来许晓昕匡华徐丽广刘丽强吴晓玲宋珊珊孙茂忠郝昌龙胡拥明
Owner JIANGNAN UNIV
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