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Hybridoma cell strain secreting procymidone monoclonal antibody and application of procymidone monoclonal antibody

A hybridoma cell line and monoclonal antibody technology, which is applied in the direction of analytical materials, biochemical equipment and methods, and material inspection products, can solve the problems of long detection time, complicated processing, and inapplicability to rapid detection of a large number of samples, and achieve good results. Effect of Assay Sensitivity and Specificity

Inactive Publication Date: 2020-02-21
江苏权正检验检测有限公司 +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, these methods have shortcomings such as complicated sample pretreatment and long detection time, and are not suitable for rapid detection of a large number of samples. In order to protect the interests of consumers, it is necessary to establish an efficient and rapid detection method for procymidone

Method used

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  • Hybridoma cell strain secreting procymidone monoclonal antibody and application of procymidone monoclonal antibody

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0043] Example 1: Synthesis of Procymidone Hapten

[0044] Since the small molecule of procymidone is not immunogenic and cannot stimulate the mice to produce an immune response and then produce antibodies, it is necessary to couple procymidone to the protein through protein linkage technology to obtain immunogenicity; protein coupling Active groups commonly used in technology include amino group, carboxyl group, hydroxyl group, mercapto group, etc. Since the molecular structure of procymidone does not contain these active groups, it needs to be derivatized.

[0045] The derived procymidone hapten structure of the present invention is as follows:

[0046] .

Embodiment 2

[0047] Example 2: Synthesis of Procymidone Complete Antigen

[0048] Weigh 2.0mg procymidone hapten (PRM-COOH), 2.1mg N-hydroxysuccinimide (NHS), dissolve in 300μL N,N-dimethylformamide (DMF), stir at room temperature for 10min; then Weigh 3.4 mg of 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (EDC), fully dissolve it in 100 μL DMF, add it to the PRM-COOH solution, and stir at room temperature to react 6 -8 h (called liquid A). Take 8mg KLH, dilute it to 4mg / mL with 0.01M carbonate buffer solution (CBS) (called solution B), then slowly add solution A to solution B drop by drop, react at room temperature overnight; then use 0.01M PBS solution Dialyzed to remove unreacted small molecular haptens, to obtain the complete antigen PRM-COOH-KLH, and identified by UV absorption scanning method.

Embodiment 3

[0049] Embodiment 3: the synthesis of procymidone coating former

[0050] Dissolve 2.2 mg of procymidone hapten (PRM-COOH) and 2.3 mg of N-hydroxysuccinimide (NHS) in 300 μL of anhydrous N,N-dimethylformamide (DMF), and stir at room temperature for 10 minutes to obtain Procymidone hapten (PRM-COOH) solution; after dissolving 3.8mg of 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (EDC) in 100μL of anhydrous DMF, add Add it to the PRM-COOH solution, stir at room temperature for 6-8 hours to obtain liquid A; dilute 10 mg of chicken ovalbumin (OVA) with 1 mL of carbonate buffer solution (CBS) with a concentration of 0.01 mmol / L to obtain liquid B; Slowly add solution A to solution B drop by drop for reaction to obtain a reaction solution; dialyze the reaction solution with PBS solution to remove unreacted small molecule haptens to obtain the original coating (PRM-COOH-OVA).

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Abstract

The invention discloses a hybridoma cell strain secreting a procymidone monoclonal antibody and application of the procymidone monoclonal antibody and belongs to the field of food security immunodetection. The hybridoma cell strain secreting the procymidone monoclonal antibody, which is disclosed by the invention, is already preserved in the China General Microbiological Culture Collection CenterCGMCC, and the preservation number is CGMCC No.17401. The procymidone monoclonal antibody secreted by the strain is applied to analysis and detection on residues of procymidone in food security detection. A procymidone monoclonal antibody cell strain disclosed by the invention can be applied to immunoassay detection, and has good detection sensitivity and specificity (the IC50 (half maximal inhibitory concentration) value is 16.58ng / mL, the cross with a procymidone analog is smaller than 10%, and the crossing-over rate=(IC50 of the procymidone / IC50 of the analog)*100%) for the procymidone.

Description

technical field [0001] The invention relates to a hybridoma cell strain secreting procymidone monoclonal antibody and application thereof, belonging to the field of food safety immunoassay. Background technique [0002] Procymidone (PRM) is a new fungicide with low toxicity and widely used in agricultural production. It mainly inhibits the synthesis of triglycerides in the bacteria, and has special effects on fungi of the genus Botrytis and Sclerotinia. Spraying procymidone at the early flowering, full flowering and fruiting stages of plants can prevent gray mold and sclerotinia of fruit trees and vegetables. However, there may be problems such as improper use of procymidone in the process of spraying, and it is easy to leave the environment in the process of spraying, so that it accumulates on plants and animals, and then enters the human body through human consumption, which is harmful to human health. affect and even threaten human life. Based on the emphasis on enviro...

Claims

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Application Information

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IPC IPC(8): C12N5/20C07K16/44C07K14/795C07D209/48G01N33/53G01N33/577
CPCC07K16/44C07K14/795C07D209/48G01N33/5308G01N33/577
Inventor 胥传来张荣荣章晓萍匡华徐丽广刘丽强宋珊珊吴晓玲胡拥明曹玉朋
Owner 江苏权正检验检测有限公司
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