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Hybridoma cell strain DCF secreting fenhexamid monoclonal antibody and application thereof

A hybridoma cell line and monoclonal antibody technology, which is applied in the field of immunoassay, can solve the problems of inapplicability to rapid detection of a large number of samples, complex processing, and long detection time

Pending Publication Date: 2022-04-12
无锡迪腾敏生物科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, these methods have disadvantages such as complicated sample pretreatment and long detection time, and are not suitable for rapid detection of a large number of samples.

Method used

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  • Hybridoma cell strain DCF secreting fenhexamid monoclonal antibody and application thereof
  • Hybridoma cell strain DCF secreting fenhexamid monoclonal antibody and application thereof
  • Hybridoma cell strain DCF secreting fenhexamid monoclonal antibody and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0055] Example 1: Synthesis of fenhexamid hapten

[0056] The structure of the fenhexamid hapten of the present invention is as follows, and the fenhexamid hapten is referred to as FH-COOH for short:

[0057]

[0058] The synthesis process of fenhexamid hapten is as follows:

[0059] Dissolve 172mg fenhexamid in 10mL acetone, add 123mg ethyl 6-bromohexanoate, 83mg anhydrous potassium carbonate, 8mg potassium iodide, reflux for 48h, filter the mixture and evaporate the acetone, dissolve the residue in 6.7mL ethanol and 3.3mL of 1M NaOH, and refluxed overnight, then acidified the solution to pHfigure 1 shown.

Embodiment 2

[0060] Example 2: Synthesis of complete antigen of fenhexamid

[0061] Weigh 8 mg fenhexamid hapten (FH-COOH), 5 mg N-hydroxysuccinimide (NHS), dissolve in 300 μL N,N-dimethylformamide (DMF), stir at room temperature for 10 min; weigh again Take 7mg of 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (EDC), fully dissolve it with 100μL DMF, add it to the FH-COOH solution, and stir at room temperature for 4-6h (called liquid A)). Take 5mg of BSA, dilute it to 2mg / 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 Dialyze to remove the unreacted small molecule hapten to obtain the complete antigen FH-COOH-BSA, the structure is as follows, and identified by UV absorption scanning method.

[0062]

Embodiment 3

[0063] Example 3: Synthesis of fenhexamid coating source

[0064] Dissolve 4 mg of fenhexamid hapten (FH-COOH) and 2.5 mg of N-hydroxysuccinimide (NHS) in 300 μL of anhydrous N,N-dimethylformamide (DMF), and stir at room temperature for 10 minutes to obtain fenhexamid hapten (FH-COOH) solution; after dissolving 4.2 mg of 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (EDC) in 100 μL of anhydrous DMF, Add it into the FH-COOH solution, stir at room temperature and react for 4-6 hours to obtain liquid A; dilute 5 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 dropwise 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 (FH-COOH-OVA).

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Abstract

The invention discloses a hybridoma cell strain DCF secreting a fenhexamid monoclonal antibody and application of the hybridoma cell strain DCF, and belongs to the technical field of immunodetection. The preservation number of the hybridoma cell strain DCF secreting the fenhexamid monoclonal antibody is CGMCC NO.45013. The fenhexamid monoclonal antibody secreted by the hybridoma cell strain DCF secreting the fenhexamid monoclonal antibody has good specificity and detection sensitivity to fenhexamid, can be used for establishing an immunological detection method of fenhexamid and detecting residues of fenhexamid in food, and has important application prospects.

Description

technical field [0001] The invention belongs to the technical field of immune detection, and in particular relates to a hybridoma cell line DCF secreting fenhexamid monoclonal antibody and an application thereof. Background technique [0002] Fenhexamid is a new systemic and protective amide fungicide developed by Bayer, which can inhibit fungal spore germination and mycelium growth, and is mainly used for botrytis cinerea and sclerotinia and black spot disease, etc., and can also show certain activity against some strains that have developed resistance. Because fenhexamid degrades quickly in the environment, is not easy to volatilize, has good compatibility with ecology, has the characteristics of safety to crops, favorable toxicological data and good ecological effects, making it an ideal comprehensive harmful substance Governance medication. At present, there are many studies on the residues of fenhexamid in food. Domestically, the residue limits in more than 20 kinds ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K16/44C12N5/20C07K14/765C07K14/77G01N33/53
Inventor 刘丽强程媛胥传来匡华徐丽广孙茂忠宋珊珊胡拥明吴爱红郭玲玲胥欣欣倪萍毕雪威郭鹏飞
Owner 无锡迪腾敏生物科技有限公司
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