Hybridoma cell strain secreting monoclone antibody of pentachlorophenol and its application

Abstract

This invention relates to a hybridoma cell line secreting a monoclonal antibody against pendimethalin and its application, belonging to the field of immunoassay. The hybridoma cell line secreting the pendimethalin monoclonal antibody of this invention was deposited on November 12, 2025, at the China General Microbiological Culture Collection Center (CGMCC), located at No. 3, Courtyard 1, Beichen West Road, Chaoyang District, Beijing, with accession number CGMCC No. 46744. The pendimethalin monoclonal antibody secreted by this hybridoma cell line is used for the analysis and detection of pendimethalin residues in food safety testing. The pendimethalin monoclonal antibody cell line obtained by this invention can be used for immunoassay detection, exhibiting good detection sensitivity and specificity (IC50) for pendimethalin. 50 The value was 1.87 ng / mL.

Description

Technical Field

[0001] This invention relates to the field of immunoassay, and in particular to a hybridoma cell line that secretes a pendimethalin monoclonal antibody and its application. Background Technology

[0002] Pencycuron, also known as pendimethalin or fenbendazole, is a non-systemic phenylurea protective fungicide. Developed jointly by Japanese and German companies, its pure form is a white crystalline powder with the chemical name 1-(4-chlorobenzyl)-1-cyclopentyl-3-phenylurea. This agent is highly effective against diseases caused by Rhizoctonia solani and is mainly used to control rice sheath blight and potato black scurf. As a protective fungicide, it is relatively stable in the environment and has low toxicity to mammals; the acute oral LD50 in rats is [not specified]. 50 A dose greater than 5000 mg / kg is relatively safe for bees.

[0003] For the detection of pendimethalin pesticide residues, high-performance liquid chromatography (HPLC), gas chromatography (GC), and GC-MS / MS are commonly used. However, these methods have drawbacks such as complex sample pretreatment and long detection times, making them unsuitable for rapid detection of large numbers of samples. To protect the interests of consumers, it is necessary to develop a highly efficient and rapid detection method for pendimethalin. Enzyme-linked immunosorbent assay (ELISA) is an extremely efficient, sensitive, and rapid detection method. It requires simple sample pretreatment, involves few purification steps, has a large analytical capacity, low detection cost, and is easy to operate, making it suitable for rapid on-site detection of large numbers of samples. Therefore, it has been widely used in pesticide residue analysis. However, the prerequisite for using ELISA to detect pendimethalin is obtaining a monoclonal antibody with high specificity and sensitivity to pendimethalin. Therefore, finding a method to prepare a monoclonal antibody with high specificity and sensitivity to pendimethalin is crucial. Summary of the Invention

[0004] To address the aforementioned technical problems, this invention provides a hybridoma cell line that secretes a monoclonal antibody against pendimethalin and its application. The monoclonal antibody secreted by this hybridoma cell line exhibits good specificity and detection sensitivity (IC50) for pendimethalin. 50 With a value of 1.87 ng / mL, it can be used to establish an immunological detection method for pendimethalin to detect pendimethalin residues in food.

[0005] This invention is achieved through the following technical solution:

[0006] The first objective of this invention is to provide a hybridoma cell line that secretes a monoclonal antibody against pendimethalin. This hybridoma cell line was deposited on November 12, 2025, at the China General Microbiological Culture Collection Center (CGMCC), located at No. 3, Courtyard 1, Beichen West Road, Chaoyang District, Beijing, with accession number CGMCC No. 46744.

[0007] In one embodiment of the present invention, the hybridoma cell line is obtained by immunizing animals with an immunogen obtained by conjugating a carrier protein with a pendimethalin hapten.

[0008] In one embodiment of the present invention, the structural formula of the pendimethalin hapten is shown below:

[0009] .

[0010] In one embodiment of the present invention, the preparation method of the pentylene hapten is as follows: tert-butyl p-aminobenzoate is coupled with (S)-1-(5-chloro-2-aminophenyl)pyrrolidine in a DCC / DMF system to obtain an amidated tert-butyl ester intermediate; the amidated tert-butyl ester intermediate is treated with TFA / DCM to remove the tert-butyl ester protecting group to obtain the pentylene hapten.

[0011] In one embodiment of the present invention, the carrier protein includes keyhole hemocyanin and / or chicken ovalbumin.

[0012] A second objective of this invention is to provide a pendimethalin monoclonal antibody secreted by the hybridoma cell line.

[0013] A third objective of this invention is to provide a composition comprising the hybridoma cell line and / or the pendimethalin monoclonal antibody.

[0014] A fourth objective of this invention is to provide a kit comprising one or more of the hybridoma cell line, the pendimethalin monoclonal antibody, and the composition described herein.

[0015] A fifth objective of this invention is to provide a test strip comprising one or more of the hybridoma cell line, the pendimethalin monoclonal antibody, and the composition described herein.

[0016] A sixth objective of this invention is to provide a chip comprising one or more of the aforementioned hybridoma cell line, the aforementioned pendimethalin monoclonal antibody, and the aforementioned composition.

[0017] A seventh objective of this invention is to provide the application of the hybridoma cell line, the pendimethalin monoclonal antibody, the composition, the kit, the test strip, or the chip in the detection of pendimethalin; the application does not relate to the diagnosis or treatment of diseases.

[0018] The eighth objective of this invention is to provide a method for preparing the hybridoma cell line that secretes the above-mentioned pendimethalin monoclonal antibody, comprising the following steps:

[0019] (1) Use pendimethalin hapten to prepare pendimethalin complete antigen, and prepare the obtained pendimethalin complete antigen into antigen-containing Freund's adjuvant and antigen-incomplete Freund's adjuvant;

[0020] (2) The Freund's adjuvant was injected subcutaneously into the back of BALB / c mice for multiple immunizations. The first immunization used complete Freund's adjuvant, and the booster immunization used incomplete Freund's adjuvant.

[0021] (3) Blood was collected from mice that had undergone the above immunization process. The serum immune titer and immunosuppressive ability of the mice were detected by indirect ELISA. Mice with high serum tebuconazole antibody content were screened to obtain immunity.

[0022] (4) The selected mice were given a final booster immunization with incomplete Freund's adjuvant, and then sprint immunization was performed by intraperitoneal injection. The sprint immunization was performed using complete pentazocone antigen without Freund's adjuvant.

[0023] (5) Fusing spleen cells and myeloma cells from BALB / c mice after sprint immunization, culturing the fused cells in a culture medium, detecting positive cell pores using indirect ELISA, and further determining the inhibitory effect of positive cell pores using indirect competitive ELISA, subcloning the positive cell pores with the best inhibition using limiting dilution method, and finally screening out hybridoma cell lines that can secrete pendimethalin monoclonal antibodies.

[0024] The molecular formula of the pendimethalin hapten mentioned in step (1) is as follows:

[0025] ;

[0026] The molecular formula of the pendimethalin complete antigen mentioned in step (1) is as follows:

[0027] .

[0028] In one embodiment of the present invention, the interval between the first immunization and the booster immunization in steps (2) and (4) is one month, the interval between booster immunizations is 21 days, and the interval between booster immunizations and the sprint immunization is 18 to 21 days.

[0029] In one embodiment of the present invention, the initial immunization dose in steps (2) and (4) is 100 μg / animal, the booster immunization dose is 50 μg / animal, and the final immunization dose is 25 μg / animal.

[0030] In one embodiment of the present invention, the immunization process in steps (2) and (4) includes one initial immunization, four booster immunizations and one sprint immunization;

[0031] In one embodiment of the present invention, the blood collection in step (3) is performed on the 7th day after the end of the 3rd immunization process.

[0032] In one embodiment of the present invention, the cell fusion in step (5) is performed 3 days after the end of the sprint immunization.

[0033] In one embodiment of the present invention, the cell fusion in step (5) is performed by the polyethylene glycol (PEG4000) method.

[0034] In one embodiment of the present invention, the culture medium in step (5) is RPMI-1640 culture medium.

[0035] In one embodiment of the present invention, the number of subclonings in step (5) is 3.

[0036] The present invention also provides the application of the above-mentioned hybridoma cell line KDZ that secretes tebuconazole monoclonal antibody or the above-mentioned method for preparing tebuconazole monoclonal antibody in the preparation of tebuconazole monoclonal antibody.

[0037] This invention provides a pendimethalin monoclonal antibody secreted by the hybridoma cell line KDZ with accession number CGMCC No. 46744.

[0038] This invention provides a method for preparing the above-mentioned pendimethalin monoclonal antibody. The method involves taking BALB / c mice, injecting paraffin oil into the peritoneum, and then injecting the hybridoma cell line KDZ with the preservation number CGMCC No. 46744 into the peritoneum. After injection, ascites fluid is collected, purified, and the obtained monoclonal antibody is stored at low temperature.

[0039] In one embodiment of the present invention, the method involves taking 8-10 week old BALB / c mice, injecting 1 mL of paraffin oil into each mouse intraperitoneally, and then injecting 1×10⁻⁶ g of paraffin oil into each mouse intraperitoneally 7 days later. 6 Ascites fluid was collected from the hybridoma cell line with accession number CGMCC No. 46744 starting from day 7. The ascites fluid was purified by the caprylic acid-ammonium sulfate method, and the obtained monoclonal antibody was stored at -20°C.

[0040] This invention provides the application of the above-mentioned pendimethalin monoclonal antibody or the above-mentioned method for preparing pendimethalin monoclonal antibody in the recognition of pendimethalin.

[0041] Compared with the prior art, the above-described technical solution of the present invention has the following advantages:

[0042] The monoclonal antibody cell line for pendimethalin obtained in this invention can be used for immunoassay detection, exhibiting good detection sensitivity and specificity (IC50) for pendimethalin. 50 The value was 1.87 ng / mL.

[0043] Preservation of biological material samples:

[0044] A hybridoma cell line KDZ secreting a monoclonal antibody against pendimethalin has been deposited at the China General Microbiological Culture Collection Center (CGMCC), located at No. 3, Courtyard 1, Beichen West Road, Chaoyang District, Beijing, Institute of Microbiology, Chinese Academy of Sciences, with accession number CGMCC No. 46744, classified as a monoclonal cell line, and deposited on November 12, 2025. Attached Figure Description

[0045] To make the content of this invention easier to understand, the invention will be further described in detail below with reference to specific embodiments and accompanying drawings.

[0046] Figure 1 This is the standard curve of the inhibition of pendimethalin monoclonal antibody against pendimethalin in this invention. Detailed Implementation

[0047] The present invention will be further described below with reference to the accompanying drawings and specific embodiments, so that those skilled in the art can better understand and implement the present invention. However, the embodiments described are not intended to limit the present invention.

[0048] Unless otherwise specified, the experimental methods used in the following examples are conventional methods, and the materials and reagents used are commercially available.

[0049] The culture media involved in the following examples are as follows:

[0050] RPMI-1640 medium (mg / L): L-arginine 290, L-asparagine 50, L-aspartic acid 20, L-cysteine ​​dihydrochloride 65.15, L-glutamic acid 20, glycine 10, L-histidine 15, L-hydroxyproline 20, L-isoleucine 50, L-leucine 50, L-lysine hydrochloride 40, L-methionine 15, L-phenylalanine 15, L-proline 20, L-serine 30, L-threonine 20, L-tryptophan 5. L-Tyrosine 23.19, L-Valine 20, Para-aminobenzoic acid 1, Calcium nitrate 100, Anhydrous magnesium sulfate 48.84, Anhydrous sodium dihydrogen phosphate 676.13, Potassium chloride 400, Sodium chloride 6000, Glucose 2000, Reduced glutathione 1, Phenol red 5, L-Glutamine 300, Biotin 0.2, D-Calcium pantothenate 0.25, Folic acid 1, I-Inositol 35, Nicotinamide 1, Choline chloride 3, Pyridoxine hydrochloride 1, Riboflavin 0.2, Thiamine hydrochloride 1, Vitamin B12 0.005, Sodium bicarbonate 2000.

[0051] The reagents involved in the following examples are as follows:

[0052] Carbonate buffer (CBS): Weigh 1.59 g of Na2CO3 and 2.93 g of NaHCO3, dissolve them separately in a small amount of double-distilled water and mix them together. Add double-distilled water to about 800 mL and mix well. Adjust the pH to 9.6 and add double-distilled water to a final volume of 1000 mL. Store at 4°C for later use.

[0053] Phosphate buffered saline (PBS): 8.00 g NaCl, 0.2 g KCl, 0.2 g KH2PO4, 2.9 g Na2HPO4·12H2O, dissolved in 800 mL pure water, pH adjusted to 7.2-7.4 with NaOH or HCl, and then brought to a final volume of 1000 mL.

[0054] PBST: PBS containing 0.05% Tween 20;

[0055] Antibody dilution solution: PBS with 0.1% gelatin added.

[0056] TMB colorimetric solution: Solution A: Na2HPO4 . 12H₂O 18.43 g, citric acid 9.33 g, diluted to 1000 mL with pure water; Solution B: 60 mg TMB dissolved in 100 mL ethylene glycol. Mix solutions A and B in a 5:1 ratio to obtain the TMB colorimetric solution, and mix fresh before use.

[0057] The detection methods involved in the following embodiments are as follows:

[0058] Method for detecting pendimethalin inhibition rate: The optimal antigen and antibody concentrations for ic-ELISA were selected using a checkerboard assay. The antigen was diluted to 0.01, 0.03, 0.1, and 0.3 μg / mL with carbonate buffer (CBS), and the antibody was diluted to 0.01, 0.03, 0.1, and 0.3 μg / mL with antibody dilution buffer. After selecting the optimal operating point, pendimethalin standards were diluted to concentrations of 0, 0.11 ng / mL, 0.33 ng / mL, 1 ng / mL, 3 ng / mL, 9 ng / mL, 27 ng / mL, and 81 ng / mL, following the ic-ELISA procedure. Finally, graphs were plotted using OriginPro 8.5 (results are shown in the figure). Figure 1 (As shown), obtain the standard inhibition curve of pendimethalin and calculate IC50. 50 .

[0059] Example 1: Synthesis of Pendimethalin Hapten

[0060] Since pendimethalin small molecules are not immunogenic and cannot stimulate mice to produce an immune response and thus antibodies, it is necessary to use protein conjugation technology to couple pendimethalin to proteins to give it immunogenicity. Commonly used active groups in protein conjugation technology include amino, carboxyl, hydroxyl, and thiol groups. Since pendimethalin does not contain amino, carboxyl, or hydroxyl groups in its molecular structure, a pendimethalin analog containing a carboxyl group is designed as its hapten.

[0061] The structure of the derivatized pendimethalin hapten (WJL-COOH) of this invention is as follows:

[0062] .

[0063] The method for preparing the pendimethalin hapten of the present invention is as follows:

[0064]

[0065] Example 2: Synthesis of Pendimethalin Complete Antigen

[0066] Weigh 10.07 mg of pendimethalin hapten (WJL-COOH) and 9.32 mg of N-hydroxysuccinimide (NHS), dissolve them in 250 μL of N,N-dimethylformamide (DMF), and stir at room temperature for 10 min. Then weigh 15.47 mg of 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (EDC), dissolve it thoroughly in 110 μL of DMF, and add it to the WJL-COOH solution. Stir at room temperature for 4-6 h (referred to as solution A). Take 6 mg of KLH, dilute it to 3 mg / mL with 0.01 M carbonate buffer (CBS) (referred to as solution B), and slowly add solution A dropwise to solution B. React at room temperature overnight. Then dialyze with 0.01 M PBS to remove unreacted small molecule hapten to obtain the complete antigen WJL-COOH-KLH, which is identified by ultraviolet absorption scanning.

[0067] Example 3: Synthesis of Pendimethalin Coating Oxide

[0068] 2.00 mg of pendimethalin hapten (WJL-COOH) and 1.85 mg of N-hydroxysuccinimide (NHS) were dissolved in 100 μL of anhydrous N,N-dimethylformamide (DMF) and reacted with stirring at room temperature for 10 min to obtain a pendimethalin hapten (WJL-COOH) solution. 3.08 mg of 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (EDC) was dissolved in 100 μL of anhydrous DMF and added to the WJL-COOH solution. The mixture was reacted with stirring at room temperature for 4-6 h to obtain solution A. 4 mg of chicken ovalbumin (OVA) was diluted with 1 mL of 0.01 mmol / L carbonate buffer (CBS) to obtain solution B. Solution A was slowly added dropwise to solution B to obtain the reaction solution. The reaction solution was dialyzed with PBS to remove unreacted small molecule hapten to obtain the coating antigen (WJL-COOH-OVA).

[0069] Example 4: Preparation of hybridoma cell lines secreting pendimethalin monoclonal antibodies

[0070] 1. Obtaining animal immunization: BALB / c mice were immunized by subcutaneous injection at multiple sites on the neck and back after mixing and emulsifying a mixture of complete pentothal antigen and an equal volume of Freund's adjuvant (except for sprint immunization). The initial immunization used complete Freund's adjuvant at a dose of 100 μg / mouse. For multiple booster immunizations, incomplete Freund's adjuvant was used at half the dose (50 μg / mouse). For sprint immunization, no adjuvant was used; the adjuvant was diluted directly with physiological saline and injected intraperitoneally at a dose halved (25 μg / mouse). The interval between the initial and second booster immunizations was one month, the interval between multiple booster immunizations was 21 days, and the interval between sprint immunization and the final booster immunization was 18-21 days. The immunization effect in mice was observed using an indirect competitive enzyme-linked immunosorbent assay (ic-ELISA), i.e., the titer and inhibition of mouse serum were detected.

[0071] 2. Cell fusion: Three days after the sprint immunization, cell fusion was performed using the standard PEG (polyethylene glycol, molecular weight 4000) method. The specific steps are as follows:

[0072] a. After euthanizing the mouse by tail dislocation and cervical dislocation, immediately disinfect the mouse in 75% alcohol for about 5 minutes. Under aseptic conditions, remove the spleen and grind it moderately with the rubber tip of a syringe and pass it through a 200-mesh cell sieve to obtain a spleen cell suspension. Collect the suspension, centrifuge (1200 rpm, 8 min), wash the spleen cells three times with RPMI-1640 medium, and after the last centrifugation, dilute the spleen cells to a certain volume, count them, and set them aside for later use.

[0073] b. Collection of SP 2 / 0 cells: 7-10 days before fusion, SP 2 / 0 tumor cells are cultured in RPMI-1640 medium containing 10% FBS (fetal bovine serum) in a 5% CO2 incubator. The required number of SP2 / 0 tumor cells before fusion is 1-4 × 10⁻⁶ cells / year. 7 To ensure that SP2 / 0 tumor cells are in the logarithmic growth phase before fusion, tumor cells are collected and suspended in RPMI-1640 basal culture medium for cell counting during fusion.

[0074] c. Fusion process (7 min): Min 1, add 1 mL of PEG 1500 to the cells dropwise from slow to fast; Min 2, let stand; Min 3 and Min 4, add 1 mL of RPMI-1640 medium dropwise over 1 min; Min 5 and Min 6, add 2 mL of RPMI-1640 medium dropwise over 1 min; Min 7, add 1 mL of RPMI-1640 medium dropwise every 10 s; then incubate at 37℃ for 5 min; centrifuge (800 rpm, 8 min), discard the supernatant, resuspend in RPMI-1640 selection medium containing 20% ​​fetal bovine serum and 2% 50×HAT, add 200 μL / well to a 96-well cell plate, and incubate at 37℃ in a 5% CO2 incubator;

[0075] 3. Cell screening and cell line establishment: On the 3rd day of cell fusion, the fused cells were screened with RPMI-1640 medium with a half-replacement. On the 5th day, the medium was completely replaced with RPMI-1640 transition medium containing 20% ​​fetal bovine serum and 1% 100×HT. On the 7th day, the cell supernatant was collected for screening.

[0076] The screening process consists of two steps: First, positive cell wells are selected using the ic-ELISA method; second, pendimethalin is used as a standard, and the inhibitory effect on positive cells is determined using the ic-ELISA method.

[0077] Select cell wells that showed good inhibition of pendimethalin standard, perform subcloning using the limiting dilution method, and test using the same method after seven days.

[0078] Three subclonings were performed using the method described above, ultimately yielding the pendimethalin monoclonal antibody cell line KDZ.

[0079] Example 5: Preparation and Identification of Pendimethalin Monoclonal Antibody

[0080] 8-10 week old BALB / c mice were injected intraperitoneally with 1 mL of sterile paraffin oil; 7 days later, each mouse was injected intraperitoneally with 1×10 6 Ascites was collected from the pentazocone hybridoma cells starting on day 7, and the ascites was purified for antibody using the caprylic acid-saturated ammonium sulfate method.

[0081] Under slightly acidic conditions, octanoic acid can precipitate other proteins in the ascites fluid besides IgG immunoglobulin. After centrifugation, the precipitate is discarded. Then, an equal volume of saturated ammonium sulfate solution is used to precipitate IgG-type monoclonal antibodies. After centrifugation, the supernatant is discarded. The antibody is dissolved in 0.01M PBS solution (pH 7.4), dialyzed to desalt, and finally purified monoclonal antibodies are obtained and stored at -20℃.

[0082] The IC50 of pendimethalin monoclonal antibody was measured using an indirect competitive ELISA. 50 The value was 1.87 ng / mL, indicating good sensitivity to pendimethalin, and it can be used for the immunoassay detection of pendimethalin.

[0083] Example 6: Application of pendimethalin monoclonal antibody

[0084] The monoclonal antibody prepared from hybridoma cell lines via in vivo ascites fluid was used in an ELISA addition and recovery assay for pendimethalin. The specific steps are as follows:

[0085] (1) Coat a 96-well microplate with 0.1 μg / mL of the coating stock diluted with carbonate buffer (CBS), 100 μL per well, at 37℃ for 2 h. Wash the plate three times with PBST washing buffer, 200 μL per well each time, for 3 min each time, and pat dry.

[0086] (2) Block with CBS containing 0.2% gelatin, 200 μL per well, block at 37℃ for 2 h, wash the plate three times with PBST washing solution, 200 μL per well each time, 3 min each time, and pat dry;

[0087] (3) Prepare 0, 0.11 ng / mL, 0.33 ng / mL, 1 ng / mL, 3 ng / mL, 9 ng / mL, 27 ng / mL and 81 ng / mL standard solutions of pendimethalin using phosphate buffer (PBS). Add the standard solutions and the extracts of the samples to be tested to the sealed microplates, 50 μL per well, and repeat each sample in 3 wells. Add 50 μL of anti-pentadiazine monoclonal antibody to each well. After reacting at 37°C for 0.5 h, wash the plate and pat dry.

[0088] (4) Add 100 μL of HRP-labeled goat anti-mouse IgG secondary antibody diluted 1:3000 with PBS containing 0.1% gelatin to each well, react at 37℃ for 0.5 h, then wash and dry the plate.

[0089] (5) Add 100 μL of TMB colorimetric solution to each well, develop the color at 37℃ for 15 min, then add 50 μL of 2M H2SO4 stop solution to each well, and measure the absorbance at 450 nm.

[0090] The standard curve of inhibition of pendimethalin monoclonal antibody against pendimethalin is shown below. Figure 1 As shown, the IC50 of pendimethalin monoclonal antibody was determined by ic-ELISA. 50 The value was 1.87 ng / mL, indicating that the antibody has good sensitivity to pendimethalin and can be used for the immunoassay detection of pendimethalin.

[0091] Obviously, the above embodiments are merely illustrative examples for clear explanation and are not intended to limit the implementation. Those skilled in the art will recognize that other variations or modifications can be made based on the above description. It is neither necessary nor possible to exhaustively list all possible implementations here. However, obvious variations or modifications derived therefrom are still within the scope of protection of this invention.

Claims

1. A hybridoma cell line secreting a monoclonal antibody against pendimethalin, characterized in that, The hybridoma cell line was deposited on November 12, 2025, at the China General Microbiological Culture Collection Center (CGMCC), located at No. 3, Courtyard 1, Beichen West Road, Chaoyang District, Beijing, with accession number CGMCC No. 46744.

2. The hybridoma cell line according to claim 1, characterized in that, The hybridoma cell line was obtained by immunizing animals with an immunogen derived from the conjugation of pendimethalin hapten with a carrier protein.

3. The hybridoma cell line according to claim 1, characterized in that, The structural formula of the pendimethalin hapten is shown below: 。 4. The hybridoma cell line according to claim 1, characterized in that, The carrier proteins include keyhole hemocyanin and / or chicken ovalbumin.

5. A pendimethalin monoclonal antibody, characterized in that, Produced by the hybridoma cell line described in any one of claims 1-4.

6. A composition, characterized in that, The composition comprises the hybridoma cell line of any one of claims 1-4 and / or the pendimethalin monoclonal antibody of claim 5.

7. A reagent kit, characterized in that, The kit comprises one or more of the hybridoma cell lines of any one of claims 1-4, the pendimethalin monoclonal antibody of claim 5, and the composition of claim 6.

8. A test strip, characterized in that, The test strip comprises one or more of the hybridoma cell lines of any one of claims 1-4, the pendimethalin monoclonal antibody of claim 5, and the composition of claim 6.

9. A chip, characterized in that, The chip comprises one or more of the hybridoma cell lines of any one of claims 1-4, the pendimethalin monoclonal antibody of claim 5, and the composition of claim 6.

10. The use of the hybridoma cell line of any one of claims 1-4, the pendimethalin monoclonal antibody of claim 5, the composition of claim 6, the kit of claim 7, the test strip of claim 8, or the chip of claim 9 in the detection of pendimethalin; the use does not relate to the diagnosis and treatment of disease.

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