A monoclonal antibody of benzimidazole drugs and application thereof

Abstract

The embodiment of the application discloses a kind of benzimidazole drug monoclonal antibodies and its application, belong to biotechnology field.A kind of benzimidazole drug monoclonal antibodies, including heavy chain variable region and light chain variable region, the amino acid sequence of CDR1, CDR2, CDR3 of the heavy chain variable region respectively as shown in SEQ ID No.5~SEQ ID No.7;The amino acid sequence of CDR1, CDR2, CDR3 of the light chain variable region respectively as shown in SEQ ID No.8~SEQ ID No.10.The application provides a kind of benzimidazole drug monoclonal antibodies with high titer, high sensitivity, and the test strip based on the antibody has higher sensitivity, provides powerful technical support for the field such as benzimidazole drug residual detection, drug research and development and quality control.

Description

Technical Field

[0001] This invention relates to the field of biotechnology, specifically to a benzimidazole monoclonal antibody and its application. Background Technology

[0002] Benzimidazole drugs are a class of broad-spectrum active compounds containing a benzimidazole core, with core representatives including albendazole, fenbendazole, mebendazole, and carbendazim, widely used in the pharmaceutical, animal husbandry, and agricultural fields. They exert their anthelmintic and bactericidal effects by targeting parasite β-tubulin and inhibiting microtubule polymerization. While possessing advantages such as a broad anthelmintic spectrum and good efficacy, misuse can easily lead to safety risks such as excessive residues, embryonic teratogenicity, liver and kidney damage, and drug resistance. my country and EU member states have set strict limits on their residue levels in food and agricultural products, necessitating highly specific and sensitive detection and application technologies.

[0003] Currently, the detection methods for benzimidazole drugs are mainly divided into instrumental analysis and immunoassay. Among instrumental analysis methods, liquid chromatography-tandem mass spectrometry (LC-MS / MS) is the gold standard for detection, possessing both high sensitivity and high specificity. However, it is expensive, requires complex pretreatment, and is time-consuming, making it unsuitable for rapid, batch detection on-site. High-performance liquid chromatography (HPLC) suffers from insufficient sensitivity and difficulty in qualitative analysis, while gas chromatography requires cumbersome derivatization and is prone to introducing errors. Other methods, such as capillary electrophoresis-mass spectrometry (CES) and fluorescence analysis, all have problems such as poor reproducibility and narrow applicability, making them difficult to apply on a large scale.

[0004] Immunoassay has become the mainstream method for on-site testing due to its simplicity, speed, and low cost, and its core relies on specific monoclonal antibodies. However, existing monoclonal antibodies against benzimidazole drugs have significant technical shortcomings: on the one hand, they lack broad-spectrum recognition, with some antibodies having narrow recognition spectra, making it difficult to simultaneously recognize benzimidazole drugs with significant structural differences, thus failing to achieve simultaneous detection of multiple commonly used benzimidazole drugs and resulting in low detection efficiency; on the other hand, they have low sensitivity, weak antibody affinity, and detection limits that cannot meet the needs of trace residue detection, making it difficult to comply with national standards and EU low residue limits.

[0005] Therefore, it is necessary to develop a broad-spectrum, highly specific, and highly sensitive monoclonal antibody against benzimidazole drugs to meet the practical needs of food safety supervision and clinical drug use monitoring. Summary of the Invention

[0006] Therefore, embodiments of the present invention provide a benzimidazole monoclonal antibody and its application.

[0007] To achieve the above objectives, the embodiments of the present invention provide the following technical solutions: According to a first aspect of the present invention, the present invention provides a benzimidazole monoclonal antibody, comprising a heavy chain variable region and a light chain variable region, wherein the amino acid sequences of CDR1, CDR2, and CDR3 of the heavy chain variable region are shown in SEQ ID No. 5 to SEQ ID No. 7, respectively; and the amino acid sequences of CDR1, CDR2, and CDR3 of the light chain variable region are shown in SEQ ID No. 8 to SEQ ID No. 10, respectively.

[0008] Further, the amino acid sequence of the heavy chain variable region is shown in SEQ ID No. 1; the amino acid sequence of the light chain variable region is shown in SEQ ID No. 2.

[0009] Furthermore, the benzimidazole monoclonal antibody is prepared using carbendazim-carrier protein conjugate as an immunogen. The preparation method of the carbendazim-carrier protein conjugate includes: 2-methoxycarbonylamino-1H-benzimidazole-6-carboxylic acid is activated by EDC / NHS and then subjected to an amidation reaction with a carrier protein to form the carbendazim-carrier protein conjugate, wherein the carrier protein is bovine serum albumin or ovalbumin.

[0010] According to a second aspect of the present invention, the present invention provides the use of the benzimidazole monoclonal antibody as described above in the preparation of products for detecting benzimidazole drugs.

[0011] Furthermore, the product is a test strip.

[0012] Furthermore, the benzimidazole drugs are carbendazim, albendazole, albendazole sulfone, albendazole sulfoxide, and albendazole-2-aminosulfone.

[0013] According to a third aspect of the present invention, the present invention provides a test strip for detecting benzimidazole drugs, wherein the conjugate pad of the test strip is coated with a benzimidazole drug monoclonal antibody as described above.

[0014] The embodiments of the present invention have the following advantages: This invention utilizes a complete antigen carbendazim-carrier protein conjugate to immunize mice, successfully obtaining a high-titer, highly sensitive monoclonal antibody against benzimidazole drugs. Based on this antibody, a highly efficient, sensitive, and specific immunoassay method for benzimidazole drugs was further established. This method not only improves detection accuracy but also demonstrates promising application prospects, providing strong technical support for the detection of benzimidazole drug residues, drug development, and quality control. Attached Figure Description

[0015] To more clearly illustrate the embodiments of the present invention or the technical solutions in the prior art, the drawings used in the description of the embodiments or the prior art will be briefly introduced below. Obviously, the drawings described below are merely exemplary, and those skilled in the art can derive other embodiments based on the provided drawings without creative effort.

[0016] Figure 1 This is an SDS-PAGE image of a benzimidazole monoclonal antibody provided by the present invention. Figure 2 This is a standard curve diagram of monoclonal antibodies against benzimidazole drugs provided by the present invention; Figure 3 The image shows the sequence identity comparison results of the heavy chain gene of the benzimidazole drug monoclonal antibody provided by this invention. Figure 4 This is a sequence identity comparison diagram of the heavy chain amino acid sequence of the benzimidazole drug monoclonal antibody provided by the present invention; Figure 5 This is a sequence identity comparison diagram of the light chain gene of the benzimidazole drug monoclonal antibody provided by the present invention; Figure 6 This is a sequence identity comparison diagram of the light chain amino acid sequence of the benzimidazole drug monoclonal antibody provided by the present invention; Figure 7 The image shows the interpretation result of the test strip provided by this invention. Detailed Implementation

[0017] The following specific embodiments illustrate the implementation of the present invention. Those skilled in the art can easily understand other advantages and effects of the present invention from the content disclosed in this specification. Obviously, the described embodiments are only some, not all, of the embodiments of the present invention. Based on the embodiments of the present invention, all other embodiments obtained by those skilled in the art without creative effort are within the scope of protection of the present invention.

[0018] Example 1: Preparation of artificial antigens for benzimidazole drugs The structural formula of the artificial antigen of benzimidazole drugs is as follows: Protein is bovine serum albumin (BSA) or ovalbumin (OVA).

[0019] The preparation method of artificial antigens for benzimidazole drugs includes the following steps: 10 mg of 2-methoxycarbonylamino-1H-benzimidazole-6-carboxylic acid and 20 mg of NHS were dissolved in 1 mL of DMF to obtain the first solution; 20 mg of EDC was dissolved in 0.1 M MES buffer (pH=5.5) to obtain the second solution; the second solution was added dropwise to the first solution and reacted at room temperature for 4 h to obtain the third solution; 30 mg of carrier protein BSA was dissolved in 0.1 M CBS (pH=9.6) to obtain the fourth solution; the third solution was added to the fourth solution, mixed well, and reacted at room temperature for another 12 h; after the reaction was completed, the reactants were placed in a dialysis bag and dialyzed with 0.01 M PBS (pH 7.2-7.4) at 4 °C for 48 h, with the dialysate changed every 12 h, to obtain the immunogen CBZ-BSA, which was aliquoted and stored at -20 °C.

[0020] The preparation method of the coating antigen CBZ-OVA is the same as that of CBZ-BSA, except that the carrier protein BSA is replaced with an equal amount of OVA.

[0021] Example 2: Preparation of monoclonal antibodies against benzimidazole drugs 1. Animal immunization Four 6-week-old Balb / c mice, numbered #1 to #4, were selected. The immunogen CBZ-BSA prepared in Example 1 was thoroughly emulsified with an equal volume of Freund's complete adjuvant and injected subcutaneously into the mice at a dose of 50 μg per mouse. Booster immunizations were performed every 2 weeks for a total of 3 booster immunizations, with the adjuvant replaced by Freund's incomplete adjuvant. The immunization method and dose were the same as the initial immunization. One week after the final immunization, tail blood was collected from the mice, and serum was separated for later use.

[0022] The antibody titer of the CBZ-OVA-coated antigen prepared in Example 1 was determined by indirect competitive ELISA to assess its specific inhibitory ability against the target drug. PBS buffer was used as a blank control, and pre-immunization mouse serum was used as a negative control. The results showed that the antibody titers of all four mice were high, reaching over 1:80,000. Among them, mouse No. 1 had the highest titer, reaching 1:160,000. Furthermore, the serum of this immunized mouse showed inhibition rates >50% for carbendazim and albendazole 50ppb standards in the dilution range of 1:20,000 to 1:80,000, indicating the production of antibodies with strong resistance. Therefore, mouse No. 1 was selected for subsequent cell fusion experiments to prepare monoclonal antibodies.

[0023] The results of the detection of immune serum titer and inhibitory ability of mouse No. 1 are shown in Table 1 below. Among them, the inhibition rate = [(OD value of titer well at the same dilution - OD value of spiked well at the same dilution) / OD value of titer well at the same dilution] × 100%.

[0024] Table 1

[0025] 2. Preparation and Identification of Monoclonal Antibodies 2.1 Monoclonal Antibody Preparation Mice were sacrificed under aseptic conditions, spleens were isolated and splenocytes were prepared. Splenocytes were fused with SP2 / 0 myeloma cells and seeded into 96-well cell culture plates. Cells were cultured in HAT selective medium, and positive wells were screened. The limiting dilution method was used to clone the positive wells, obtaining and establishing a hybridoma cell line that stably secretes monoclonal antibodies against benzimidazole drugs. Eight-week-old Balb / c mice were pretreated with 0.8 mL of sterile paraffin oil via intraperitoneal injection. Seven days later, each mouse was intraperitoneally injected with 5 × 10⁶ 7E4B11 hybridoma cells in logarithmic growth phase. 5 One per animal; 10 days later, ascites fluid was collected and purified using the caprylic acid-saturated ammonium sulfate method to obtain benzimidazole monoclonal antibodies.

[0026] 2.2 Monoclonal antibody purity identification The purified monoclonal antibody was analyzed by SDS-PAGE electrophoresis, such as... Figure 1 As shown, the purified antibody lanes exhibited only two characteristic bands: the IgG heavy chain (50 kDa) and the light chain (25 kDa), with no obvious impurities. Quantification by grayscale analysis confirmed that the purified antibody purity was ≥95%.

[0027] 2.3 Monoclonal antibody sensitivity detection The detection sensitivity of the purified monoclonal antibody was analyzed using an indirect competitive ELISA method: CBZ-OVA prepared in Example 1 was used as the coating antigen, and a series of concentration gradients (0, 0.5, 1, 1.5, 2, 2.5, 3.0 ng / mL) of carbendazim, albendazole, albendazole sulfone, albendazole sulfoxide, and albendazole-2-aminosulfone standards were added respectively. Subsequent ELISA procedures were followed for color development and OD value measurement. The inhibition rate at each concentration was calculated, and the results are shown in Table 2 below. A standard curve was plotted. Figure 2 ), calculate the half-maximal inhibitory concentration (IC50) 50 ).

[0028] Table 2

[0029] The results showed that the IC50 values ​​of carbendazim, albendazole, albendazole sulfone, albendazole sulfoxide, and albendazole-2-aminosulfone were... 50 The concentrations were 1.42 ng / mL, 1.48 ng / mL, 1.86 ng / mL, 2.40 ng / mL, and 2.37 ng / mL, respectively, with a limit of detection of 0.5 ng / mL, indicating that the monoclonal antibody can detect the above-mentioned benzimidazole drugs with high sensitivity.

[0030] 2.4 Monoclonal antibody specificity analysis Other non-benzimidazole drugs, such as ivermectin and gentamicin, were detected using the indirect competitive ELISA method described in section 2.3, and the IC50 values ​​of each drug were obtained. 50 Using carbendazim as the reference (cross-reactivity rate = 100%), the cross-reactivity rate (CR) of each drug was calculated using the following formula: CR = (Carbendazim IC50) 50 / Other drugs IC 50 ) × 100%.

[0031] The results of the cross-reactivity tests for each drug are shown in Table 3 below.

[0032] Table 3

[0033] The results showed that the monoclonal antibody exhibited a cross-reactivity of 96%-100% with carbendazim and albendazole, demonstrating high recognition ability; the cross-reactivity with albendazole sulfone, albendazole sulfoxide, and albendazole-2-aminosulfone was 60%-76%, indicating strong recognition ability for the main metabolites of these drugs, covering the detection of their metabolite residues; the cross-reactivity with non-benzimidazole drugs such as ivermectin and gentamicin was <0.1%, indicating that the antibody has extremely high specificity and the detection results are not easily interfered with by other drugs, making it suitable for the accurate detection of specific benzimidazole drug residues in actual samples such as livestock and poultry meat, aquatic products, and feed.

[0034] Example 3: Cloning of light and heavy chain variable region genes of benzimidazole monoclonal antibodies (1) Hybridoma cell culture and total RNA extraction Hybridoma cells were cultured in RPMI 1640 complete medium at 37°C with 5% CO2. Total RNA was extracted from the cells using a total RNA extraction kit.

[0035] (2) Synthesis of the first strand of cDNA Takara reverse transcription kit synthesizes cDNA.

[0036] (3) Gene amplification Design downstream primers and upstream universal primers for Lambda, Kappa, and Heavy chains.

[0037] Upstream universal primer F: AAGCGTGGTATCAACGCAGA; Light chain downstream primer R κ :AACATTGATGTCTTTGGGGTAGAA; Light chain λ downstream primer Rλ :AATCGTACACACCAGTGTGTGGG; Heavy chain downstream primer R H :AGGGATCCAGAGTTCCAGGT.

[0038] PCR was performed using the first strand of cDNA as a template in a 50 μl reaction volume.

[0039] PCR reaction system: template 3μl, upstream primer (10μM) 2.5μl, downstream primer (10μM) 2.5μl, 2×Taq enzyme 25μl, sterile water 17μl.

[0040] The landing PCR reaction conditions were as follows: 98℃ for 30 seconds; 98℃ for 15 seconds, 64℃-58℃ for 30 seconds, decreasing by 0.5℃ each time until reaching 58℃, for 10 cycles; 72℃ for 30 seconds; 98℃ for 15 seconds, 56℃ for 30 seconds, 72℃ for 30 seconds, for 15 cycles; and the program was terminated at 72℃ for 7 minutes.

[0041] (4) Cloning and screening of PCR amplification products The PCR products were subjected to 1.5% agarose gel electrophoresis. The Kappa, Lambda, and Heavy chain fragments of the antibody were recovered using a PCR product recovery kit. The fragments were inserted into the pLB vector using a pLB zero-background rapid cloning kit and transformed into DH5α competent cells (ampicillin-resistant). Recombinant positive clones were screened and sequenced.

[0042] The amino acid sequence of the heavy chain variable region of the benzimidazole monoclonal antibody is shown in SEQ ID No. 1, and the amino acid sequence of the light chain variable region is shown in SEQ ID No. 2; the nucleotide sequence encoded by the heavy chain variable region is shown in SEQ ID No. 3, and the nucleotide sequence encoded by the light chain variable region is shown in SEQ ID No. 4.

[0043] The sequences of SEQ ID No. 1 to SEQ ID No. 4 are as follows: EVKLVESGGGLVKPGGSLKLTCAASGFTFSSYAMSWVRQIPDKRLEWVASISSGGNTYYPDSVKGRFIISRDNARNILNLQMNSLKSEDTAMYYCARPLDYGFKKNWYFDVWGAGTTVTVSS (SEQ ID No. 1); DIVMTQSTSSLSASLGDRVTISCRASQDISNYLNWYQQKPDGTVKLLIYYTSRLHSGVPARFSGSGSGTDYSLTISNLEQEDVATYFCQQGNALWTFGGGTKLEIK (SEQ ID No.2); GAAGTGAAGCTGGTGGAGTCTGGGGGAGGCTTAGTGAAGCCTGGAGGGTCCCTGAAACTCACCTGTGCAGCCTCTGGATTCACTTTCAGTAGCTATGCCATGTCTTGGGTTCGCCAGATTCCAGACAAGAGGCTGGAGTGGGTCGCATCCATTAGTAGTGGTGGTAACACCTACTATCCAGACAGTGTGAAGGGCCGGTTCATCATCTCCAGAGATAATGCCAGGAACATCCTGAACCTGCAAATGAACAGTCTAAAGTCTGAGGACACGGCCATGTATTACTGTGCAAGACCATTGGACTACGGTTTTAAGAAGAACTGGTATTTCGATGTCTGGGGCGCAGGGACCACGGTCACCGTCTCCTCA (SEQ ID No.3); GATATTGTGATGACACAGTCTACATCCTCCCTGTCTGCCTCTCTGGGAGACAGAGTCACCATCAGTTGCAGGGCAAGTCAGGACATTAGTAATTATTTAAACTGGTATCAGCAGAAACCAGATGGAACTGTTAAACTCCTGATCTATTACACATCAAGGTTACACTCAGGAGTCCCAGCAAGGTTCAGTGGCAGTGGGTCTGGAACAGACTATTCTCTCACCATTAGCAACCTGGAACAAGAAGATGTTGCCACTTACTTTTGCCAACAGGGTAATGCGCTGTGGACGTTCGGTGGAGGCACCAAGCTGGAAATCAAA (SEQ ID No.4).

[0044] (5) Variable region nucleotide and amino acid sequences and homology analysis BLAST alignment analysis in the NCBI database showed that the monoclonal antibody heavy chain variable region gene sequence had the highest sequence identity with the mouse immunoglobulin heavy chain variable region mRNA (Sequence ID: AJ851868.3), with a sequence identity of 281 / 291 (97%). Figure 3 As shown. The amino acid sequence of the variable region of the monoclonal antibody heavy chain has the highest identity with the amino acid sequence of the variable region of the mouse immunoglobulin heavy chain (Sequence ID: AAN39095.1), with a sequence identity of 96 / 112 (86%). Figure 4 As shown.

[0045] The gene sequence of the light chain variable region of this monoclonal antibody showed the highest sequence identity with the mRNA of the variable region of the light chain of mouse immunoglobulin κ (Sequence ID: X55042.1), with a sequence identity of 301 / 310 (97%). Figure 5 As shown. The amino acid sequence of the variable region of the monoclonal antibody light chain showed the highest identity with the mouse immunoglobulin κ light chain (Sequence ID: pirS69901), with a sequence identity of 10¹ / 10⁶ (95%). Figure 6 As shown.

[0046] The results of gene sequence and amino acid sequence identity analysis of the light chain and heavy chain variable regions encoding benzimidazole monoclonal antibodies showed that no sequences identical to those of the present invention were found.

[0047] The Novopro online CDR annotation tool was used to analyze the CDR regions of light chain variable region and heavy chain variable region sequences to obtain their CDR regions.

[0048] The amino acid sequences of CDR1, CDR2, and CDR3 in the heavy chain variable region of the benzimidazole monoclonal antibody are shown in SEQ ID No. 5 to SEQ ID No. 7, respectively; the amino acid sequences of CDR1, CDR2, and CDR3 in the light chain variable region are shown in SEQ ID No. 8 to SEQ ID No. 10, respectively.

[0049] The sequences of SEQ ID No. 5 to SEQ ID No. 10 are as follows: GFTFSSYA (SEQ ID No. 5); ISSGGNT (SEQ ID No. 6); ARPLDYGFKKNWYFDV(SEQ ID No.7); QDISNY (SEQ ID No. 8); YTS (SEQ ID No. 9); QQGNALWT (SEQ ID No. 10).

[0050] Example 4: Preparation and application of colloidal gold test strips for benzimidazole drugs 1. Preparation of colloidal gold immunochromatographic test strips for benzimidazole drugs (1) Preparation of gold-labeled pads: Colloidal gold particles with a particle size of 20 nm were mixed with monoclonal antibodies of benzimidazole drugs at a ratio of 10 μg antibody / mL colloidal gold solution. The pH was adjusted to 8.3. The mixture was sprayed onto a polyester film to form a uniform gold-labeled layer. The sprayed polyester film was dried at 37°C for 2 h to obtain the gold-labeled pads. (2) Processing of NC membrane: Detection lines (T) and control lines (C) are pre-marked on the NC membrane with a spacing of 5 mm. Antigen CBZ-OVA prepared in Example 1 is sprayed on the T line, and goat anti-rabbit IgG is sprayed on the C line. The marked NC membrane is dried at 37°C for 2 hours and then set aside. (3) Assembly of test strips: Paste the sample pad, gold label pad, NC membrane and absorbent pad in sequence onto the PVC base plate, ensuring that there is appropriate overlap between each component to form a complete liquid flow channel. Cut, package and seal the assembled test strips.

[0051] Result determination: Both the C and T lines on the test strip show color, with the T line showing much stronger color than the C line. The result is negative (-), indicating that the sample does not contain benzimidazole drugs or the color is much lower than the detection limit. If the C line of the test strip shows color, and the T line shows the same color as the C line, the T line shows a weaker color than the C line, or the T line does not show color, the result is positive (+), indicating that the concentration of benzimidazole drugs in the sample is equal to or higher than the detection limit. If neither the C line nor the T line of the test strip shows color, it indicates improper operation or that the test strip has expired.

[0052] 2. Performance evaluation of colloidal gold immunochromatographic test strips for benzimidazole drugs (1) Sensitivity The standard solutions of benzimidazole drugs at different concentrations were tested, and the results are shown in Table 4 below.

[0053] Table 4

[0054] The results showed that the benzimidazole colloidal gold immunochromatographic test strip prepared in this invention has high sensitivity for the detection of benzimidazole drugs, with a detection limit of 3 ng / mL for carbendazim and albendazole, and a detection limit of 5 ng / mL for albendazole sulfone, albendazole sulfoxide, and albendazole-2-aminosulfone.

[0055] (2) Specificity Using the benzimidazole colloidal gold immunochromatographic test strip prepared according to the present invention, tests were conducted on carbendazim, albendazole, albendazole sulfone, albendazole sulfoxide, albendazole-2-aminosulfone, ivermectin, and gentamicin (all at a concentration of 10 ng / mL). The results showed that the target benzimidazole drugs carbendazim, albendazole, albendazole sulfoxide, and albendazole-2-aminosulfone were all positive, while the other non-target drugs ivermectin and gentamicin were negative. This indicates that the test strip provided by the present invention does not cross-react with other drugs and has good specificity.

[0056] Although the present invention has been described in detail above with general descriptions and specific embodiments, modifications or improvements can be made to it, which will be obvious to those skilled in the art. Therefore, all such modifications or improvements made without departing from the spirit of the present invention fall within the scope of protection claimed by the present invention.

Claims

1. A monoclonal antibody against a benzimidazole drug, characterized in that, It includes a heavy chain variable region and a light chain variable region, wherein the amino acid sequences of CDR1, CDR2, and CDR3 of the heavy chain variable region are shown in SEQ ID No. 5 to SEQ ID No. 7, respectively; The amino acid sequences of CDR1, CDR2, and CDR3 in the light chain variable region are shown in SEQ ID No. 8 to SEQ ID No. 10, respectively.

2. The benzimidazole monoclonal antibody according to claim 1, characterized in that, The amino acid sequence of the heavy chain variable region is shown in SEQ ID No. 1; the amino acid sequence of the light chain variable region is shown in SEQ ID No.

2.

3. The benzimidazole monoclonal antibody according to claim 1, characterized in that, The benzimidazole monoclonal antibody is prepared using carbendazim-carrier protein conjugate as an immunogen. The preparation method of the carbendazim-carrier protein conjugate includes: 2-methoxycarbonylamino-1H-benzimidazole-6-carboxylic acid is activated by EDC / NHS and then subjected to amidation reaction with a carrier protein to form the carbendazim-carrier protein conjugate, wherein the carrier protein is bovine serum albumin or ovalbumin.

4. The use of a benzimidazole monoclonal antibody as described in claim 1 in the preparation of products for detecting benzimidazole drugs.

5. The application according to claim 4, characterized in that, The product in question is a test strip.

6. The application according to claim 4, characterized in that, The benzimidazole drugs mentioned are carbendazim, albendazole, albendazole sulfone, albendazole sulfoxide, and albendazole-2-aminosulfone.

7. A test strip for detecting benzimidazole drugs, characterized in that, The conjugation pad of the test strip is coated with a benzimidazole monoclonal antibody as described in claim 1.

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