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Triadimefon monoclonal antibody hybridoma cell strain B11S and application thereof

A technology of hybridoma cell lines and monoclonal antibodies, which is applied in the direction of animal/human protein, serum albumin, ovalbumin, etc., can solve the problems of high cost, complicated processing, and many interferences, and achieve good specificity, The effect of good detection sensitivity

Inactive Publication Date: 2020-06-19
JIANGNAN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0004] In order to effectively supervise and monitor the use of triadimefon in food, a method with good specificity and high sensitivity is needed. However, current detection methods such as high performance liquid chromatography, high performance liquid chromatography tandem mass spectrometry, etc., sample pretreatment process It is more complex, expensive, and there are many interfering substances in food, so the instrument method is not suitable for on-site detection

Method used

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  • Triadimefon monoclonal antibody hybridoma cell strain B11S and application thereof
  • Triadimefon monoclonal antibody hybridoma cell strain B11S and application thereof
  • Triadimefon monoclonal antibody hybridoma cell strain B11S and application thereof

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Embodiment 1

[0024] Example 1 Preparation of a triadimefon monoclonal antibody hybridoma cell line B11S

[0025] (1) Synthesis of hapten:

[0026]

[0027] The synthesis scheme of the triadimefon hapten is shown above. The derivation steps are briefly described as follows: Dissolve sodium chloroacetate and anhydrous sodium carbonate in 2 mL of pure water to obtain solution A, and dissolve triadimenol, a structural analogue of triadimefon, in 95% ethanol to obtain solution B. Then, liquid A was slowly added to liquid B, and stirred at room temperature for 10 h to obtain liquid C. The reactant was filtered, and the filtrate was evaporated to dryness in a water bath at 80°C. The precipitated TDF was then stored at 4°C for use.

[0028] (2) Preparation of complete antigen:

[0029] Preparation of immunogen TDF-BSA: Weigh 6.3mg TDF, 10.3mg 1-ethylcarbodiimide hydrochloride, 6.2mg N-hydroxysuccinimide, add 400μL anhydrous N,N-dimethylformaldehyde The amide was dissolved (referred to as l...

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Abstract

The invention discloses a triadimefon monoclonal antibody hybridoma cell strain B11S and application thereof, and belongs to the technical field of food safety immunodetection. The triadimefon monoclonal antibody hybridoma cell strain B11S disclosed by the invention is preserved in China General Microbiological Culture Collection Center (CGMCC), is classified and named as a monoclonal cell strain,is preserved on October 14, 2019, and has a preservation number of CGMCC No. 18518. A BALB / c mouse is immunized by a triadimefon complete antigen. High-titer and low-IC50 mouse splenocytes are takenand fused with myeloma cells through a PEG method, and the hybridoma cell strain B11S is obtained through screening of an indirect competitive enzyme-linked immunosorbent assay and three times of subcloning. A monoclonal antibody secreted by the cell strain has good specificity and detection sensitivity (IC50 value is 5.25 ng / mL) to triadimefon, provides a raw material for immunodetection of triadimefon residues in foods, and has practical application value.

Description

technical field [0001] The invention relates to a triadimefon monoclonal antibody hybridoma cell line B11S and an application thereof, belonging to the technical field of food safety immunoassay. Background technique [0002] Triadimefon (TDF) is a high-efficiency, broad-spectrum azole fungicide with preventive, therapeutic and eradicating effects, and strong systemic effect (can conduct up and down). It has special effects on powdery mildew, rust, and smut, and its bactericidal effect is to inhibit the biosynthesis of ergosterol. It is mainly used to prevent and control diseases of wheat, fruit trees, vegetables, melons, flowers and other crops. [0003] "GB 2763-2016 Maximum Residue Limits of Pesticides in Food" stipulates the maximum residue limit (MRL) of triadimefon in various foods, and its content in wheat (except wheat) and dry grain (except corn) The maximum residue limit standard (MRL) is 0.2mg / kg, the maximum residue limit standard (MRL) in solanaceous vegetable...

Claims

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Application Information

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IPC IPC(8): C12N5/20C07K16/44C07K14/765C07K14/77C07D249/08G01N33/53C12R1/91
CPCC07D249/08C07K14/765C07K14/77C07K16/44G01N33/5308
Inventor 匡华林璐胥传来徐丽广孙茂忠刘丽强吴晓玲宋珊珊胡拥明郝昌龙
Owner JIANGNAN UNIV
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