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Hybridoma cell strain FQ-B12, anti-thiabendazole monoclonal antibody produced thereby and application of hybridoma cell strain FQ-B12

A technology of FQ-B12 and hybridoma cell lines, applied in the direction of microorganisms, biochemical equipment and methods, peptides, etc., can solve the problems of complex processing, high sensitivity, high cost, etc., and achieve the effect of high antibody specificity

Inactive Publication Date: 2015-01-21
JIANGSU ACADEMY OF AGRICULTURAL SCIENCES
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

These methods have low detection limits and high sensitivity, and relevant technical standards are available for reference, but due to the need for expensive instruments, specialized operators, and the complexity, high cost, and long time of sample pretreatment, they cannot better meet the requirements. Quick and easy on-site inspection requirements

Method used

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  • Hybridoma cell strain FQ-B12, anti-thiabendazole monoclonal antibody produced thereby and application of hybridoma cell strain FQ-B12
  • Hybridoma cell strain FQ-B12, anti-thiabendazole monoclonal antibody produced thereby and application of hybridoma cell strain FQ-B12
  • Hybridoma cell strain FQ-B12, anti-thiabendazole monoclonal antibody produced thereby and application of hybridoma cell strain FQ-B12

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Experimental program
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Effect test

Embodiment 1

[0038] The detection of embodiment 1 thiabendazole standard substance

[0039] The indirect competitive ELISA method was used to conduct a preliminary exploration of the detection curve. Methods as below:

[0040] Coating: Dilute the coating antigen (H3-OVA) to 5 μg mL with coating buffer (0.05mol / L, pH9.6) -1 Add to the microplate, 50 μl per well, and incubate in a 37°C incubator for 2 hours;

[0041] (2) Plate washing: wash 5 times with washing solution PBST (0.05% Tween 20, 0.01mol / L, pH7.4), and pat dry with absorbent paper;

[0042] (3) Blocking: add 200 μL of 1% gelatin to each well, and incubate at 37° C. for 1.5 h;

[0043] (4) Plate washing: same as (2);

[0044] (5) Add pesticide and antibody mixture: Dilute thiabendazole standard solution 3 times with PBST phosphate buffer solution, add 25 μL to each well, then add 25 μL of diluted antibody solution, incubate at 37°C for 1 hour, wash with PBST 5 times, And set positive control and negative control in parallel; ...

Embodiment 2

[0051] Detect thiabendazole residue in embodiment 2 apple, pear sample

[0052] H3-OVA-coated ELISA plate 50 μL / well, thiabendazole monoclonal antibody diluted 1600 times as the working concentration, 1% gelatin as the blocker, PBST phosphate buffer pH value 7.5, ionic strength 0.8M, organic solvent For methanol, 50 μL / well in the microtiter plate.

[0053] Sample preparation to be tested:

[0054] Apple and pear samples were purchased from Nanjing Suguo Supermarket. Weighed 10g of pre-homogenized samples, added standard substance (thiabendazole) to a final concentration of 200ng / g, mixed well, left at room temperature for 1h, added 10mL of methanol, and fully Shake, 4000rpm, centrifuge for 5min, and transfer the supernatant into a 25mL volumetric flask. Add 10mL of methanol to the precipitate, shake fully, centrifuge at 4000rpm for 5min, transfer the supernatant into a 25mL volumetric flask, and dilute to 25mL with the optimal working buffer (without organic solvent). Take...

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Abstract

The invention relates to a hybridoma cell strain FQ-B12, an anti-benzimidazole sterilizing agent namely a thiabendazole monoclonal antibody secreted by the cell strain, and an application of the hybridoma cell strain FQ-B12. The hybridoma cell strain FQ-B12 is collected in China Center for Type Culture Collection (CCTCC), and has the collection number of CCTCC NO. C201491. The monoclonal antibody secreted by the hybridoma cell strain FQ-B12 can be used for identifying thiabendazole, is high in sensitivity and good in specificity, and has the 50% inhibition concentration (IC50) of 6.0ng.mL<-1> to thiabendazole. An enzyme-linked immunosorbent assay method researched by using the monoclonal antibody can achieve the detection of thiabendazole in water, apples and pears.

Description

technical field [0001] The invention relates to a hybridoma cell strain FQ-B12, a monoclonal antibody secreted by the hybridoma cell line against thiabendazole, a fungicide of benzimidazoles, and its application. Background technique [0002] Benzimidazole pesticides are an important class of therapeutic fungicides. Due to their remarkable efficacy and low toxicity, they are widely used as broad-spectrum systemic fungicides in the production of fruits infected by fungi. According to the "2007 Pesticide Management Information Compilation", carbendazim, benomyl and thiabendazole are the main types of benzimidazole pesticides. The main toxicity of this kind of pesticide is teratogenic effect, which was first proposed by Lapras M et al. in 1973. It has a long residual period, slow degradation in the natural environment, and has a certain enrichment effect. And consumption of this kind of substance reaches a certain dose when it is harmful to the human body. [0003] my countr...

Claims

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Application Information

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IPC IPC(8): C07K16/44C12N5/20
Inventor 刘凤权王利民蔡佳
Owner JIANGSU ACADEMY OF AGRICULTURAL SCIENCES