Sulfoxaflor hapten, hybridoma cell line, antibody and use thereof

Abstract

The present invention provides a sulfoxaflor hapten, a hybridoma cell line, an antibody and its application, which belongs to the field of immunochemistry technology. The present invention provides a sulfoxaflor hapten, a complete antigen and a hybridoma cell line that secretes a sulfoxaflor monoclonal antibody, which has been deposited in the General Microbiology Center of the China Culture Collection of Microorganisms (CGMCC), the deposit address of which is the Institute of Microbiology, Chinese Academy of Sciences, No. 3, Yard 1, Beichen West Road, Chaoyang District, Beijing. It is classified and named as a monoclonal cell line, the deposit date is April 18, 2024, and the deposit number is CGMCC No. 45912. The monoclonal antibody secreted by this cell line has good detection sensitivity for sulfoxaflor and can be used for residual detection of sulfoxaflor.

Description

Technical Field

[0001] The present invention relates to the field of food safety immunoassay, and in particular to sulfoxaflor hapten, hybridoma cell line, antibody and application thereof. Background Art

[0002] Sulfoxaflor's common name is Sulfoxaflor, and its chemical name is [Methyl(oxo){1-[6-(trifluoromethyl)-3-pyridyl]ethyl}-γ-6-sulfanyl]cyanamide. Its mechanism of action is through its action on the insect nervous system, activating unique binding sites within the nicotinic acetylcholine receptor (NACRT). Its primary uses are for controlling a variety of piercing-sucking pests in cotton, rapeseed, fruit trees, soybeans, fruit, small grains, vegetables, rice, lawns, and ornamental plants, including aphids, stink bugs, stink bugs, whiteflies, scales, planthoppers, certain psyllids, and thrips. It is effective against piercing-sucking pests resistant to nicotine, pyrethroid, organophosphate, and carbamate pesticides, making it a preferred agent for integrated pest management.

[0003] Currently, the detection of sulfoxaflor is primarily based on instrumental methods, with gas chromatography, liquid chromatography, and gas chromatography-mass spectrometry being the most common. While these chromatography-based methods offer high sensitivity and specificity, they have drawbacks, such as the need for thorough sample cleanup, high solvent consumption, expensive equipment, and the need for skilled technicians. Therefore, a rapid and simple method for detecting sulfoxaflor residues is needed.

[0004] Enzyme-linked immunosorbent assay (ELISA) is a highly efficient, sensitive and rapid detection method. During detection, the pre-treatment of the sample is simple, the purification step is few, the analytical capacity is large, the detection cost is low and it is easy to operate. It is suitable for on-site rapid detection of a large number of samples and is therefore widely used in drug residue analysis. The premise for using enzyme-linked immunosorbent assay to detect sulfoxaflor is to obtain a monoclonal antibody with high specificity and high sensitivity to sulfoxaflor, so it is very important to find a method for preparing a monoclonal antibody with high specificity and high sensitivity to sulfoxaflor. The inventor attempts to prepare sulfoxaflor monoclonal antibody by hybridoma cells, but in the process of preparing the hybridoma cell line that can secrete sulfoxaflor monoclonal antibody, how to prepare sulfoxaflor hapten and complete antigen and how to make mice produce strong immune effect, further research is needed; how to make the hybridoma cell line prepared can successfully secrete sulfoxaflor monoclonal antibody, further research is needed; how to make the sulfoxaflor monoclonal antibody secreted highly specific and highly sensitive, further research is also needed. Summary of the Invention

[0005] To solve the above technical problems, the present invention provides a sulfoxaflor hapten, a hybridoma cell line, an antibody and its application. The sulfoxaflor monoclonal antibody secreted by the hybridoma cell line has a good detection sensitivity (IC 50 The value was 4.58 ng / mL), which can be used to establish an immunological detection method for sulfoxaflor and detect the residues of sulfoxaflor in food.

[0006] The technical solutions of the present invention are as follows:

[0007] The first object of the present invention is to provide a sulfoxaflor hapten, the structural formula of the sulfoxaflor hapten is:

[0008]

[0009] The second object of the present invention is to provide a complete sulfoxaflor antigen, which is prepared from the sulfoxaflor hapten and a carrier protein.

[0010] In one embodiment of the present invention, the carrier protein is selected from keyhole limpet hemocyanin and / or chicken ovalbumin.

[0011] The third object of the present invention is to provide the use of the sulfoxaflor hapten in preparing a hybridoma cell line that secretes sulfoxaflor monoclonal antibodies.

[0012] The fourth object of the present invention is to provide a hybridoma cell line that secretes a monoclonal antibody against sulfoxaflor, wherein the hybridoma cell line is obtained by immunizing mice with the sulfoxaflor hapten by synthesizing a complete sulfoxaflor antigen; the hybridoma cell line has been deposited in the General Microbiology Center of the China Culture Collection of Microorganisms (CGMCC), with the deposit address being the Institute of Microbiology, Chinese Academy of Sciences, No. 3, Yard 1, Beichen West Road, Chaoyang District, Beijing, and is classified and named a monoclonal cell line. The deposit date is April 18, 2024, and the deposit number is CGMCC No. 45912.

[0013] The fifth object of the present invention is to provide a sulfoxaflor monoclonal antibody secreted and produced by the hybridoma cell line.

[0014] The sixth object of the present invention is to provide a composition comprising the hybridoma cell line and / or the sulfoxaflor monoclonal antibody.

[0015] The seventh object of the present invention is to provide a kit, which contains at least one of the hybridoma cell line, the sulfoxaflor monoclonal antibody and the composition.

[0016] An eighth object of the present invention is to provide a test strip comprising at least one of the hybridoma cell line, the sulfoxaflor monoclonal antibody, and the composition.

[0017] The ninth object of the present invention is to provide use of the hybridoma cell line, the sulfoxaflor monoclonal antibody, the composition, the kit or the test strip in detecting sulfoxaflor.

[0018] The present invention also provides a method for preparing the hybridoma cell line secreting the sulfoxaflor monoclonal antibody, comprising the following steps:

[0019] Step 1: preparing sulfoxaflor hapten and sulfoxaflor complete antigen, and emulsifying the obtained sulfoxaflor complete antigen with Freund's adjuvant or incomplete Freund's adjuvant to prepare an immunogen;

[0020] Step 2: The obtained immunogen was injected subcutaneously into the back of BALB / c mice for multiple immunizations, with complete Freund's adjuvant used for the first immunization and incomplete Freund's adjuvant used for the booster immunization;

[0021] Step 3: Blood was collected from the mice that had undergone the above immunization process, and the immune titer and immunosuppressive capacity of the mouse serum were detected by indirect ELISA to screen out mice with high sensitivity of sulfoxaflor antibodies in the serum;

[0022] Step 4: The screened mice were immunized by intraperitoneal injection with complete sulfoxaflor antigen without Freund's adjuvant;

[0023] Step 5: Spleen cells from BALB / c mice after sprint immunization are fused with myeloma cells. The fused cells are screened and cultured in HAT medium. Positive cell wells are detected by indirect ELISA, and the inhibitory effect of the positive cell wells is further determined by indirect competitive ELISA. The positive cell wells with the best inhibition are subcloned by limiting dilution, and finally, a hybridoma cell line that secretes a highly sensitive monoclonal antibody against sulfoxaflor is screened.

[0024] In step 1, the structural formula of the sulfoxaflor hapten is as follows:

[0025]

[0026] The structural formula of the sulfoxaflor complete antigen 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 the booster immunizations is 21 days, and the interval between the booster immunization and the sprint immunization is 18 to 21 days.

[0029] In one embodiment of the present invention, the first immunization dose in steps 2 and 4 is 100 μg / animal, the booster immunization dose is 50 μg / animal, and the sprint 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 third 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 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 subcloning in step 5 is 4 times.

[0036] The present invention provides a method for preparing the above-mentioned sulfoxaflor monoclonal antibody. The method comprises taking BALB / c mice, intraperitoneally injecting paraffin oil, and then intraperitoneally injecting a hybridoma cell line with a deposit number of CGMCC No. 45912, collecting ascites after the injection, purifying the ascites, and cryopreserving the obtained monoclonal antibody.

[0037] Beneficial effects of the present invention:

[0038] (1) The monoclonal antibody against sulfoxaflor obtained by the present invention has a good detection sensitivity (IC 50 value was 4.58 ng / mL);

[0039] (2) The sulfoxaflor monoclonal antibody cell line obtained in the present invention can be used for immunoassay detection.

[0040] Deposit of biological materials:

[0041] A hybridoma cell line GLY that secretes a monoclonal antibody against sulfoxaflor has been deposited in the General Microbiology Center of the China Culture Collection Administration, with the deposit address being the Institute of Microbiology, Chinese Academy of Sciences, No. 3, Yard 1, Beichen West Road, Chaoyang District, Beijing. It is classified and named a monoclonal cell line, with a deposit date of April 18, 2024, and a deposit number of CGMCC No. 45912. BRIEF DESCRIPTION OF THE DRAWINGS

[0042] In order to make the content of the present invention more clearly understood, the present invention is further described in detail below based on specific embodiments of the present invention in conjunction with the accompanying drawings, wherein:

[0043] Figure 1 This is the standard curve of the inhibition of sulfoxaflor monoclonal antibody against sulfoxaflor. DETAILED DESCRIPTION

[0044] 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 the present invention and implement it. However, the embodiments are not intended to limit the present invention.

[0045] The culture medium involved in the following examples is as follows:

[0046] RPMI-1640 medium (mg / L): L-arginine 290, L-asparagine 50, L-aspartic acid 20, L-cystine 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, p-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.

[0047] The reagents involved in the following examples are as follows:

[0048] Carbonate buffer (CBS): Weigh 1.59 g of Na2CO3 and 2.93 g of NaHCO3, dissolve them in a small amount of double-distilled water, mix them, add double-distilled water to about 800 mL, adjust the pH to 9.6, add double-distilled water to 1000 mL, and store at 4°C until used.

[0049] 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, adjusted to pH 7.2–7.4 with NaOH or HCl, and made up to 1000 mL.

[0050] PBST: PBS containing 0.05% Tween 20;

[0051] Antibody diluent: PBS containing 0.1% gelatin;

[0052] TMB color development solution: Solution A: Na2HPO 4. 18.43g of 12H2O, 9.33g of citric acid, and dilute to 1000mL with pure water. Solution B: Dissolve 60mg of TMB in 100mL of ethylene glycol. Mix Solution A and Solution B in a ratio of 5:1 to create the TMB colorimetric solution. Re-mix before use.

[0053] The detection methods involved in the following embodiments are as follows:

[0054] Method for detecting the inhibition rate of sulfoxaflor: The most appropriate antigen and antibody concentrations for the IC-ELISA were selected using a checkerboard test. The antigen was diluted to 0.01, 0.03, 0.1, and 0.3 μg / mL using carbonate buffer (CBS), and the antibody was diluted to 0.03, 0.1, 0.3, and 1 μg / mL using antibody diluent. After selecting the optimal working point, the sulfoxaflor standard was diluted to 8 concentrations (0, 0.37, 1.111, 3.333, 10, 30, 90, and 270 ng / mL). The IC-ELISA procedure was followed, and the graph was generated using OriginPro 8.5 (the results are shown in the figure). Figure 1 As shown), obtain the standard inhibition curve of sulfoxaflor and calculate IC 50 .

[0055] Example 1: Synthesis of sulfoxaflor hapten.

[0056] Since the small molecule of sulfoxaflor is not immunogenic and cannot stimulate mice to produce an immune response and then produce antibodies, sulfoxaflor needs to be coupled to the protein through protein coupling technology to make it immunogenic; the active groups commonly used in protein coupling technology include amino, carboxyl, hydroxyl, thiol, etc. Since the molecular structure of sulfoxaflor does not contain these active groups, sulfoxaflor is derivatized.

[0057] A mixture of sulfoxaflor (200 mg) and aqueous sodium hydroxide solution (10%, 10 mL) was heated in an oil bath at reflux temperature under magnetic stirring for 4.5 hours. After the solution was cooled to room temperature, it was extracted with ether to remove non-acidic substances. Aqueous hydrochloric acid solution (15%, 50 mL) was added in batches. The formed suspension was poured into a separatory funnel and extracted with ether (3×50 mL). The organic extract was washed with water and dried over sodium sulfate. After filtration, the solvent was removed by rotary evaporation, and the yellow liquid residue was distilled under reduced pressure to obtain a sulfoxaflor derivative (73 mg), which was a sulfoxaflor hapten.

[0058] The structural formula of the sulfoxaflor hapten is as follows:

[0059]

[0060] Example 2: Synthesis of complete sulfoxaflor antigen.

[0061] 6 mg of sulfoxaflor hapten and 4.4 mg of N-hydroxysuccinimide (NHS) were dissolved in 200 μL of N,N-dimethylformamide (DMF) and stirred at room temperature for 10 minutes. 7.5 mg of 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (EDC) was then added to the sulfoxaflor hapten solution and stirred at room temperature for 6-8 hours for activation. 6 mg of keyhole limpet hemocyanin (KLH) was added to 3 mL of 0.01 M carbonate buffer (CBS) and thoroughly dissolved. The activated hapten was then slowly added to the KLH solution and stirred at room temperature overnight. The hapten was then dialyzed against 0.01 M PBS to remove unreacted small molecules, resulting in a relatively pure complete antigen, which was then identified by UV absorption scanning.

[0062] Since the protein molecular weight of KLH is larger than that of BSA, its immunogenicity is stronger, and it has more sites that can be coupled with haptens, so it has a stronger ability to recognize the detected drugs and a wider detection linear range.

[0063] Example 3: Synthesis of sulfoxaflor coating.

[0064] Dissolve 3.2 mg of sulfoxaflor hapten and 2.4 mg of N-hydroxysuccinimide (NHS) in 200 μL of anhydrous N,N-dimethylformamide (DMF) and stir at room temperature for 10 minutes. Dissolve 4.2 mg of 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (EDC) in this solution and stir at room temperature for 6-8 hours to obtain a hapten activation solution. Dissolve 6 mg of chicken ovalbumin (OVA) in carbonate buffer (CBS). Slowly add the hapten activation solution to the protein dilution solution and stir at room temperature overnight. Then, dialyze the reaction solution against 0.01 M PBS to remove unreacted small molecules to obtain the coating source.

[0065] Example 4: Preparation of hybridoma cell lines secreting sulfoxaflor monoclonal antibodies.

[0066] 1. Acquisition of animal immunity

[0067] After emulsification, sulfoxaflor complete antigen was mixed with an equal amount of Freund's adjuvant, and BALB / c mice were immunized by subcutaneous injection at multiple points on the back of the neck (except for booster immunization). Complete Freund's adjuvant was used for the first immunization at a dose of 100 μg per mouse. Incomplete Freund's adjuvant was used for multiple booster immunizations, and the dose was halved to 50 μg per mouse. The booster immunization was performed without adjuvant and directly diluted with normal saline before intraperitoneal injection, and the dose was further halved to 25 μg per mouse. The interval between the first and second booster immunizations was one month, between multiple booster immunizations was 21 days, and between the booster immunization and the final booster immunization was 18-21 days. The immune response of the mice was assessed by indirect competitive enzyme-linked immunosorbent assay (IC-ELISA), i.e., the titer and inhibitory effect of the mouse serum were measured.

[0068] In addition, the present invention also uses BSA to prepare complete antigens, but the titer and inhibition of mouse serum are not as good as the results of the complete antigen prepared by KLH.

[0069] 2. Cell fusion

[0070] Three days after the sprint immunization, cell fusion was performed according to the conventional PEG (polyethylene glycol, molecular weight 4000) method. The specific steps are as follows:

[0071] a. Blood was collected by tail amputation. Mice were sacrificed by cervical dislocation and immediately disinfected in 75% alcohol for about 5 minutes. The spleen of the mouse was removed aseptically. The spleen was moderately ground with the rubber tip of a syringe and passed through a 200-mesh cell sieve to obtain a splenocyte suspension. The suspension was collected and centrifuged (1200 rpm, 8 minutes). The splenocytes were washed three times with RPMI-1640 medium. After the final centrifugation, the splenocytes were diluted to a certain volume, counted, and set aside.

[0072] b. Collect SP2 / 0 cells: 7-10 days before fusion, SP2 / 0 tumor cells were expanded in RPMI-1640 medium containing 10% FBS (fetal bovine serum) in a 5% CO2 incubator. Before fusion, the number of SP2 / 0 tumor cells should reach 1-4×10 7 , ensure that SP2 / 0 tumor cells are in the logarithmic growth phase before fusion. When fusion occurs, tumor cells are collected, suspended in RPMI-1640 basal culture medium, and cell counts are performed;

[0073] c. Fusion process (7 min): In the first minute, add 1 mL of PEG 4000 dropwise to the cells in a gradual, increasing order. In the second minute, allow the cells to stand. In the third and fourth minutes, add 1 mL of RPMI-1640 medium dropwise over 1 minute. In the fifth and sixth minutes, add 2 mL of RPMI-1640 medium dropwise over 1 minute. In the seventh minute, add 1 mL of RPMI-1640 medium dropwise every 10 seconds. Except for the second minute, shake the solution continuously. Then, incubate at 37°C for 5 minutes. Centrifuge (800 rpm, 8 minutes), discard the supernatant, and resuspend the cells in RPMI-1640 selection medium supplemented with 20% fetal bovine serum and 2% 50× HAT. Plate 200 μL / well of the cells into a 96-well plate and incubate in a 37°C, 5% CO2 incubator.

[0074] 3. Cell screening and cell line establishment

[0075] On the third day after cell fusion, the fused cells were subjected to a half-change of RPMI-1640 screening culture medium. On the fifth day, the culture medium was fully replaced with RPMI-1640 transition culture medium containing 20% ​​fetal bovine serum and 1% 100×HT. On the seventh day, the cell supernatant was collected for screening.

[0076] The screening was divided into two steps: the first step was to screen out the positive cell wells using the ic-ELISA method, and the second step was to use sulfoxaflor as the standard and determine the inhibitory effect on the positive cells using the ic-ELISA method.

[0077] The cell wells with better inhibition against the sulfoxaflor standard were selected and subcloned using the limiting dilution method. The cells were then tested using the same method seven days later.

[0078] Subcloning was performed at least three times according to the above method to finally obtain a sulfoxaflor hybridoma cell line.

[0079] Example 5: Preparation and identification of sulfoxaflor monoclonal antibodies.

[0080] 8-10 week old BALB / c mice were injected intraperitoneally with 1 mL of sterile paraffin oil. Seven days later, 1×10 6The ascites of the sulfoxaflor hybridoma cells were collected from the seventh day and the antibodies were purified by the octanoic acid-saturated ammonium sulfate method.

[0081] Under slightly acidic conditions, octanoic acid can precipitate other miscellaneous proteins in the ascites except IgG immunoglobulin, and then centrifuge and discard the precipitate; then use an equal amount of saturated ammonium sulfate solution to precipitate IgG type monoclonal antibodies, centrifuge, discard the supernatant, dissolve it with 0.01M PBS solution (pH 7.4), dialyze and desalt, and finally obtain the purified monoclonal antibody and store it at -20°C.

[0082] The IC of the monoclonal antibody against sulfoxaflor was measured using an indirect competitive ELISA. 50 The value was 4.58 ng / mL, indicating that it had good sensitivity to sulfoxaflor and could be used for sulfoxaflor immunoassay detection.

[0083] Example 6: Application of sulfoxaflor monoclonal antibody.

[0084] The monoclonal antibody prepared from the hybridoma cell line in vivo using ascites was applied to the ELISA spike-recovery test of sulfoxaflor. The specific steps are as follows:

[0085] (1) Coat a 96-well microtiter plate with 100 μL of the coating agent diluted in carbonate buffer (CBS) at a concentration of 0.1 μg / mL per well. Bake at 37°C for 2 h. Wash the plate three times with PBST, 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, bake at 37°C for 2 h, wash the plate three times with PBST, 200 μL per well each time, 3 min each time, and pat dry;

[0087] (3) Prepare 0 ng / mL, 0.37 ng / mL, 1.111 ng / mL, 3.333 ng / mL, 10 ng / mL, 30 ng / mL, 90 ng / mL, and 270 ng / mL sulfoxaflor standard solutions in phosphate buffered saline (PBS). Add 50 μL of the standard solutions and the sample extracts to the blocked ELISA plate, repeating the process for each sample in three wells. Then, add 50 μL of sulfoxaflor monoclonal antibody diluted to 0.1 μg / mL to each well. Incubate at 37°C for 30 min, then wash the plate and pat dry.

[0088] (4) Add 100 μL of HRP-labeled goat anti-mouse IgG secondary antibody diluted 1:3000 in PBS containing 0.1% gelatin to each well, react at 37°C for 30 min, and then wash the plate and pat dry.

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

[0090] (6) Addition recovery and sample pretreatment:

[0091] Rice was selected as the test sample.

[0092] The sample to be tested was crushed and passed through a 20-mesh standard sieve. Three samples were weighed, each 20 g, and 5 ppb, 10 ppb, and 50 ppb of sulfoxaflor standard were added to the samples (according to the linear range of the antibody and IC 50 Set the spiked concentration), add 10 mL of water, vortex mix thoroughly, let stand for 30 minutes, and filter. Add 50 mL of acetone to the sample, shake on an electric shaker for 30 minutes, filter through rapid qualitative filter paper, and transfer to a beaker. The residue is extracted once again with 30 mL of acetone according to the above method. Wash the residue twice with 30 mL of acetone. Combine the washings into a beaker, concentrate to near dryness in a 50°C water bath, and reconstitute with 5 mL of 10% acetone-PBS solution (i.e., diluted fivefold to reduce the influence of the sample matrix).

[0093] The recovery rates of the spiked products were 104%, 96% and 106% respectively in the indirect competitive ELISA test.

[0094] Obviously, the above embodiments are merely examples for clarity of explanation and are not intended to limit the implementation methods. Those skilled in the art will appreciate that other variations or modifications can be made based on the above description. It is not necessary and impossible to enumerate all implementation methods here. Obvious variations or modifications arising therefrom remain within the scope of protection of the present invention.

Claims

1. A sulfoxaflor hapten, characterized in that: The structural formula of the sulfoxaflor hapten is:

2. A complete sulfoxaflor antigen, characterized in that: The method is prepared from the sulfoxaflor hapten according to claim 1 and a carrier protein.

3. The complete sulfoxaflor antigen according to claim 2, characterized in that The carrier protein is selected from keyhole limpet hemocyanin and / or chicken ovalbumin.

4. Use of the sulfoxaflor hapten according to claim 1 in preparing a hybridoma cell line secreting a sulfoxaflor monoclonal antibody.

5. A hybridoma cell line secreting a monoclonal antibody against sulfoxaflor, characterized in that: The hybridoma cell line is obtained by immunizing mice with the sulfoxaflor hapten described in claim 1 by synthesizing the sulfoxaflor complete antigen; the hybridoma cell line has been deposited in the General Microbiology Center of the China Culture Collection of Microorganisms (CGMCC), the deposit address is No. 3, Yard 1, Beichen West Road, Chaoyang District, Beijing, Institute of Microbiology, Chinese Academy of Sciences, classified and named as a monoclonal cell strain, the deposit date is April 18, 2024, and the deposit number is CGMCC No. 45912.

6. A sulfoxaflor monoclonal antibody, characterized in that: Secreted and produced by the hybridoma cell line according to claim 5.

7. A composition, characterized in that The composition contains the hybridoma cell line according to claim 5 and / or the sulfoxaflor monoclonal antibody according to claim 6.

8. A kit, characterized in that The kit contains at least one of the hybridoma cell line according to claim 5, the sulfoxaflor monoclonal antibody according to claim 6, and the composition according to claim 7.

9. A test strip, characterized in that: The test strip contains at least one of the hybridoma cell line according to claim 5, the sulfoxaflor monoclonal antibody according to claim 6, and the composition according to claim 7.

10. Use of the hybridoma cell line according to claim 5, the sulfoxaflor monoclonal antibody according to claim 6, the composition according to claim 7, the kit according to claim 8 or the test strip according to claim 9 in detecting sulfoxaflor.

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