Hybridoma cell strain secreting terodiline monoclonal antibody and its application

Abstract

The present application relates to a hybridoma cell strain secreting terodiline monoclonal antibody and its application, and belongs to the technical field of immune detection. The present application uses 3,5-dinitrosalicylic acid as a hapten to prepare a complete antigen for immunizing a host, and a hybridoma cell strain secreting terodiline monoclonal antibody is screened and obtained, which is preserved in the China General Microbiological Culture Collection Center with a preservation number of CGMCC NO. 46230. The monoclonal antibody secreted by the hybridoma has high specificity and detection sensitivity for terodiline, with an IC 50 value of 2.14 ng / mL, and no cross reaction with terodiline structural analogs, and can be applied to the preparation of terodiline detection products, providing a powerful tool for related research and application.

Description

Technical Field

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

[0002] Teroxyphenate, a dinitrophenol herbicide, is a contact herbicide that affects mitochondrial and chloroplast respiration and photosynthesis. In mitochondria, at low concentrations, it acts as an uncoupling agent for oxidative phosphorylation; at higher concentrations, it may inhibit the electron transport chain before cytochrome c. It is commonly used to control annual broadleaf weeds in various crops. The structure of dinitrophenol compounds is extremely dangerous even at very low concentrations, especially to animals, and can cause developmental toxicity. It is toxic upon skin contact and highly toxic if ingested. Due to its long persistence, teroxyphenate can accumulate in soil. Furthermore, it may cause long-term adverse effects in aquatic environments. In China, the maximum residue limit for teroxyphenate is 0.01 mg / kg. Detecting the presence of teroxyphenate residues in agricultural products is crucial for protecting human health.

[0003] Currently, the detection methods for terfenadine are mainly instrumental, with commonly used methods including high-performance liquid chromatography (HPLC), liquid chromatography-tandem mass spectrometry (LC-MS / MS), high-performance liquid chromatography-mass spectrometry (HPLC-MS), and ultra-high-performance liquid chromatography-tandem mass spectrometry (UPLC-MS / MS). For example, patent CN114487224A discloses a method for detecting acidic pesticides in fruits and vegetables, using solid-phase extraction-high-performance chromatography-tandem mass spectrometry for qualitative and quantitative analysis of 12 acidic pesticide residues in fruits and vegetables. Although these chromatographic methods have high sensitivity and specificity, they also have some drawbacks, such as the need for thorough sample purification, high solvent consumption, expensive equipment, and skilled technicians. Therefore, a rapid and simple method for detecting terfenadine residues is needed.

[0004] 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 is widely used in drug residue analysis. However, the prerequisite for using ELISA to detect telexambin is obtaining a monoclonal antibody with high specificity and sensitivity to telexambin. Therefore, finding a method to prepare a monoclonal antibody with high specificity and sensitivity to telexambin is crucial. The inventors attempted to prepare telexambin monoclonal antibodies using hybridoma cells. However, further research is needed on how to prepare telexambin hapten and complete antigen, how to induce a strong immune response in mice, how to ensure the successful secretion of telexambin monoclonal antibodies by the prepared hybridoma cell lines, and how to ensure that the secreted telexambin monoclonal antibodies have high specificity and sensitivity. Summary of the Invention

[0005] To address the aforementioned technical problems, this invention provides a hybridoma cell line that secretes telexamyl monoclonal antibody and its application. This invention uses 3,5-dinitrosalicylic acid as a hapten to prepare a complete antigen for immunizing the host, and screens to obtain a hybridoma cell line that secretes telexamyl monoclonal antibody. This cell line is deposited at the China General Microbiological Culture Collection Center (CGMCC) with accession number CGMCC NO. 46230.

[0006] A hybridoma cell line secreting telexambin monoclonal antibody, the hybridoma cell line having the accession number CGMCC NO.46230.

[0007] Furthermore, the method for preparing the above-mentioned hybridoma cell line includes the following steps:

[0008] Step S1: Prepare telexin hapten 1 and telexin complete antigen. Emulsify the obtained telexin complete antigen with Freund's adjuvant or incomplete Freund's adjuvant to prepare an immunogen.

[0009] Step S2: The obtained immunogen is injected subcutaneously into BALB / c mice via the back for multiple immunizations. The first immunization uses complete Freund's adjuvant, and the booster immunization uses incomplete Freund's adjuvant.

[0010] Step S3: Prepare telexin hapten 2 and telexin coated antigen. Collect blood from mice that have undergone the above immunization process. Detect the serum immunotiter and immunosuppressive ability of mice by indirect ELISA. Screen mice with high sensitivity of telexin antibody in serum.

[0011] Step S4: The selected mice are immunized by intraperitoneal injection using telexin complete antigen without Freund's adjuvant.

[0012] Step S5: Fuse spleen cells and myeloma cells from BALB / c mice after sprint immunization. The fused cells are then screened and cultured in HAT medium. Positive cell pores are detected using indirect ELISA, and the inhibitory effect of positive cell pores is further determined using indirect competitive ELISA. The positive cell pores with the best inhibition are subcloned using limiting dilution, and finally, hybridoma cell lines that can secrete highly sensitive telexamyl monoclonal antibodies are screened out.

[0013] Furthermore, the structural formula of the telexambin hapten 1 is as follows:

[0014] Furthermore, the structural formula of the telexambin hapten 2 is as follows:

[0015]

[0016] A second objective of this invention is to provide the application of the above-mentioned hybridoma cell line in the preparation of telexambin monoclonal antibodies.

[0017] A third objective of this invention is to provide a telexin monoclonal antibody, wherein the telexin monoclonal antibody is secreted by the aforementioned hybridoma cell line.

[0018] A fourth object of the present invention is to provide a composition comprising the above-described hybridoma cell line and / or the above-described monoclonal antibody.

[0019] A fifth object of the present invention is to provide the use of the above-mentioned hybridoma cell line, the above-mentioned telexin monoclonal antibody, or the above-mentioned composition in the preparation of telexin detection products.

[0020] Furthermore, the telexin detection products include reagents, test strips, kits, or biochips.

[0021] A sixth object of the present invention is to provide a test strip containing the above-mentioned hybridoma cell line, the above-mentioned telexambin monoclonal antibody, or the above-mentioned composition.

[0022] Furthermore, the test strip also includes a sample pad, a conjugate pad, an absorbent pad, and a cellulose acetate membrane. The conjugate pad is coated with telexin monoclonal antibody, which is labeled with colloidal gold or fluorescent microspheres.

[0023] A seventh object of the present invention is to provide a kit containing the above-described hybridoma cell line, the above-described telexambin monoclonal antibody, and the above-described composition.

[0024] Furthermore, the kit also includes an ELISA plate, telexin-coated antigen, telexin standard solution, enzyme-labeled secondary antibody, and chromogenic solution. In one embodiment of the invention, the enzyme-labeled secondary antibody is labeled with horseradish peroxidase.

[0025] In one embodiment of the present invention, the color developing solution is a TMB color developing solution.

[0026] An eighth object of the present invention is to provide the use of the above-described hybridoma cell line, the above-described telexin monoclonal antibody, the above-described composition, the above-described test strip, or the above-described kit in the detection of telexin.

[0027] The beneficial effects of this invention are:

[0028] The hybridoma cell line provided by this invention can secrete monoclonal antibodies against telexamb, providing a stable antibody source for the specific detection of telexamb. The produced monoclonal antibody exhibits high specificity and sensitivity in the detection of telexamb, with an IC50 value of [missing information - likely related to IC50]. 50 With a value of 2.14 ng / mL, various telexambin detection products can be developed using the above-mentioned hybridoma cell lines and monoclonal antibodies or their combinations, such as reagents, test strips, kits, and biochips. These products can meet the needs of telexambin detection in different occasions, providing an efficient, accurate, and convenient means for the specific detection of telexambin, with significant technical effects and wide application value.

[0029] Preservation of biological materials

[0030] A hybridoma cell line secreting telexambin monoclonal antibody has been deposited at the China General Microbiological Culture Collection Center (CGMCC), Institute of Microbiology, Chinese Academy of Sciences, No. 3, No. 1 Beichen West Road, Chaoyang District, Beijing. It is classified as a monoclonal cell line, deposited on October 31, 2024, with accession number CGMCC NO.46230. Attached Figure Description

[0031] 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, wherein...

[0032] Figure 1 This is the LC-MS / MS characterization diagram of telexambin hapten 2 in Example 1 of the present invention;

[0033] Figure 2 This is the inhibition standard curve of the telexin monoclonal antibody in Example 5 of the present invention. Detailed Implementation

[0034] 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.

[0035] The culture medium involved in the examples is shown below:

[0036] 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.

[0037] The reagents involved in the culture medium are shown below:

[0038] Carbonate buffer (CBS): Weigh 1.59g of Na2CO3 and 2.93g of NaHCO3, dissolve them separately in a small amount of double-distilled water and mix them together. Add double-distilled water to about 800mL and mix well. Adjust the pH to 9.6 and add double-distilled water to a final volume of 1000mL. Store at 4℃ for later use.

[0039] Phosphate-buffered saline (PBS): 8.00g NaCl, 0.2g KCl, 0.2g KH2PO4, 2.9g Na2HPO4·12H2O, dissolved in 800mL pure water, pH adjusted to 7.2-7.4 with NaOH or HCl, and then brought to a final volume of 1000mL.

[0040] PBST: PBS containing 0.05% Tween 20;

[0041] Antibody dilution buffer: PBS containing 0.1% gelatin;

[0042] TMB colorimetric solution: Solution A: 18.43g Na2HPO4·12H2O, 9.33g citric acid, diluted to 1000mL with pure water; Solution B: 60mg TMB dissolved in 100mL ethylene glycol. Mix solutions A and B in a 5:1 ratio to obtain the TMB colorimetric solution. Mix again before use.

[0043] The detection methods involved in the embodiments are as follows:

[0044] Method for detecting telexin 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 0.03, 0.1, 0.3, and 1 μg / mL with antibody dilution buffer. After selecting the optimal operating point, telexin standard was diluted to eight concentrations (0, 0.069, 0.21, 0.62, 1.85, 5.56, 16.7, and 50 ng / mL). Following the ic-ELISA procedure, the results were plotted using OriginPro 8.5 (see results below). Figure 1 (As shown), the standard inhibition curve of telexambin was obtained, and the IC50 was calculated. 50 .

[0045] Example 1: Synthesis of Terliphenol Hapten

[0046] Since terephthalene is a small molecule and lacks immunogenicity, it cannot stimulate an immune response in mice to produce antibodies. Therefore, it is necessary to use protein conjugation technology to couple terephthalene to a protein to acquire immunogenicity. Commonly used active groups in protein conjugation technology include amino, carboxyl, hydroxyl, and thiol groups. However, since the terephthalene molecule does not contain these active groups, a terephthalene derivatization was designed.

[0047] Terliphenol hapten 1 is 3,5-dinitrosalicylic acid (CAS: 609-99-4).

[0048] Derivatization of telexambin hapten 2:

[0049] First, 2-hydroxy-5-tert-butyl-1,3-dinitrobenzene (1.8 g, 7.6 mmol) was dissolved in toluene (20 mL) and mixed with N,N-dimethylformamide (DMF) (5 mL) and thionyl chloride (SOCl2); 1.6 mL, 22.8 mmol). The mixture was then stirred at 0 °C for 6 h, and rotary evaporated to obtain a pale yellow solid. The precipitate was washed with water and filtered to obtain product A. Product A was then dissolved in tetrahydrofuran (10 mL) and mixed with (iPr)2EtN (1.9 g, 15.2 mmol). Methyl 4-aminobutyrate (1.3 g, 11.4 mmol) was dissolved in water, the pH was adjusted to 8.5, and the solution of product A was slowly added at 0 °C for 6 h. Rotary evaporated to obtain a yellow solid. Finally, the residue was dissolved in ethyl acetate (50 mL) and washed with HCl (20 mL, 0.1 mol / L) and water (20 mL). The resulting mixture was stirred at 75°C for 7 h, then concentrated to obtain a pale yellow viscous substance. It was treated with water and extracted with ethyl acetate, then dried over NaSO4, concentrated on a rotary evaporator, and purified on a silica gel column to obtain a pale yellow solid. The pale yellow solid was dissolved in anhydrous methanol (20 mL) at 25°C, and NaOH (2 M, 20 mL) was slowly added. The mixture was stirred for 10 min and heated to 45°C overnight. The mixture was filtered through a diatomaceous earth mat, and the solid was washed with ethyl acetate. The filtrate was concentrated under reduced pressure, and the residue was dissolved in ethyl acetate (100 mL) and washed successively with water (3 × 100 mL) and brine (100 mL). The organic layer was separated, dried over sodium sulfate, filtered, and concentrated under vacuum to obtain the terephthalic acid derivative (60 mg, 8%). The LC-MS / MS characterization results of terephthalic acid hapten 2 are as follows: Figure 1 As shown.

[0050] Example 2: Synthesis of Terliphenol Complete Antigen

[0051] 6 mg of telexin hapten 1 and 4.4 mg of N-hydroxysuccinimide (NHS) were weighed and dissolved in 200 μL of N,N-dimethylformamide (DMF), and the mixture was stirred at room temperature for 10 min. 7.5 mg of 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (EDC) was weighed and added to the telexin hapten solution, and the mixture was stirred at room temperature for 6-8 h for activation. 6 mg of keyhole hemocyanin (KLH) was added to 3 mL of 0.01 M carbonate buffer (CBS) and dissolved thoroughly. The activated hapten was slowly added to the diluted KLH solution, and the mixture was stirred overnight at room temperature. The solution was then dialyzed with 0.01 M PBS to remove unreacted small molecules, yielding a relatively pure complete antigen, which was identified by UV absorption scanning. The structural formula of the telexin complete antigen is shown below:

[0052]

[0053] Example 3: Synthesis of telexin coating agent

[0054] 3.2 mg of terephthalamide hapten 2 and 2.4 mg of N-hydroxysuccinimide (NHS) were dissolved in 200 μL of anhydrous N,N-dimethylformamide (DMF) and reacted with stirring at room temperature for 10 min. 4.2 mg of 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (EDC) was dissolved in the above solution and reacted with stirring at room temperature for 6-8 h to obtain the hapten activation solution. 6 mg of chicken ovalbumin (OVA) was dissolved in carbonate buffer (CBS). The hapten activation solution was slowly added to the protein dilution solution and stirred overnight at room temperature. Then, the reaction solution was dialyzed against 0.01 M PBS to remove unreacted small molecules, yielding the coating antigen. The structural formula of the terephthalamide coating antigen is shown below:

[0055]

[0056] Example 4: Preparation of hybridoma cell lines secreting telexambin monoclonal antibodies

[0057] 1. Acquisition of immunity in animals

[0058] After emulsifying a mixture of complete telexin antigen and an equal volume of Freund's adjuvant, BALB / c mice were immunized by subcutaneous injection at multiple sites on the back of the neck (except for sprint immunization). The first immunization used complete Freund's adjuvant at a dose of 100 μg / mouse. For multiple booster immunizations, incomplete Freund's adjuvant was used at a dose halved to 50 μg / mouse. For sprint immunization, no adjuvant was used; the mixture was diluted directly with physiological saline and injected intraperitoneally at a dose halved again to 25 μg / mouse. The interval between the first and second booster immunizations was one month, the interval between multiple booster immunizations was 21 days, and the interval between sprint immunizations and the last booster immunization was 18-21 days. The immunization effect of mice was observed by indirect competitive enzyme-linked immunosorbent assay (ic-ELISA), that is, by detecting the titer and inhibition of mouse serum, and mice with high sensitivity of telexin antibody in serum were screened.

[0059] 2. Cell fusion

[0060] Three days after the sprint immunization, cell fusion was performed using the standard PEG (polyethylene glycol, molecular weight 4000) method, with the following specific steps:

[0061] 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 minutes), 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.

[0062] b. Collection of SP2 / 0 cells: 7-10 days before fusion, SP2 / 0 tumor cells are expanded in RPMI-1640 medium containing 10% FBS (fetal bovine serum) in a 5% CO2 incubator. The number of SP2 / 0 tumor cells should reach 1-4 × 10⁶ cells / year before fusion. 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.

[0063] c. Fusion process (7 min): At min 1, add 1 mL of PEG 4000 to the cells dropwise, gradually increasing the speed; at min 2, allow to stand; at min 3 and min 4, add 1 mL of RPMI-1640 medium dropwise over 1 min; at min 5 and min 6, add 2 mL of RPMI-1640 medium dropwise over 1 min; at min 7, add 1 mL of RPMI-1640 medium dropwise every 10 s. Except for min 2, continuously agitate the solution. Then incubate at 37°C for 5 min; centrifuge (800 rpm, 8 min), discard the supernatant, and 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°C in a 5% CO2 incubator.

[0064] 3. Cell selection and cell line establishment

[0065] On day 3 after cell fusion, the fused cells were partially replaced with RPMI-1640 selection medium. On day 5, the medium was completely replaced with RPMI-1640 transition medium containing 20% ​​fetal bovine serum and 1% 100×HT. On day 7, the cell supernatant was collected for screening.

[0066] The screening process consists of two steps: First, positive cell wells are selected using the ic-ELISA method. Second, terlipine is used as a standard, and the inhibitory effect on positive cells is determined using the ic-ELISA method.

[0067] Cell wells that showed good inhibition of telexin standard were selected, and subcloning was performed using the limiting dilution method. The cells were then tested using the same method after seven days.

[0068] At least three subcloning operations were performed using the method described above to finally obtain the telexin monoclonal antibody cell line.

[0069] Example 5: Preparation and Identification of Terliphenol Monoclonal Antibody

[0070] 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 fluid was collected from telexin hybridoma cells starting on day 7, and the ascites fluid was purified for antibody using the caprylic acid-saturated ammonium sulfate method.

[0071] 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 antibodies are dissolved in 0.01M PBS solution (pH=7.4), dialyzed to desalt, and finally purified monoclonal antibodies are obtained and stored at -20℃.

[0072] The standard inhibition curve of telexambin monoclonal antibody was obtained using an indirect competitive ELISA, as follows: Figure 2 As shown, telexambin exhibits good detection sensitivity (IC50). 50 The values ​​were 2.14 ng / mL, which can be used for the immunoassay of telexambin.

[0073] Cross-reactivity experiments were conducted on monoclonal antibodies with structural analogues of terbuprofen (pentanoyl nitrophenol, abamectin, sec-butylamine, fluroxypyr, and flubendioxonil). The results showed that the absorbance values ​​of the enzyme-labeled plate for quizalofop-p-ethyl in each well did not differ significantly with the dilution concentration of the standard. Specific results are shown in Table 1. This indicates that the monoclonal antibody has a very low cross-reactivity rate to other structural analogues but good sensitivity to terbuprofen. The cross-reactivity rate (%) was calculated as follows: (Teraprofen IC50) / (Teraprofen IC50) = ... 50 ) / (Similar IC 50 )×100%.

[0074] Table 1 Cross-reactivity rates of telexin monoclonal antibodies to telexin structural analogues

[0075]

[0076] Example 6: Application of Terliphenol Monoclonal Antibody

[0077] The monoclonal antibody prepared from hybridoma cell lines via in vivo ascites fluid was used in an ELISA addition and recovery assay for telexambin. The specific steps are as follows:

[0078] (1) Coat a 96-well microplate with 0.1 μg / mL of the coating stock diluted with carbonate buffer (CBS), 100 μL per well, dry at 37°C 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.

[0079] (2) Block with CBS containing 0.2% gelatin, 200 μL per well, dry at 37°C 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;

[0080] (3) Prepare 0, 0.037, 0.111, 0.333, 1, 3, 9 and 27 ng / mL telexin standard solutions with 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. Then add 50 μL of telexin monoclonal antibody diluted to 0.1 μg / mL to each well. After reacting at 37℃ for 30 min, wash the plate and pat dry.

[0081] (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°C for 30 min, then wash the plate and pat dry.

[0082] (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.

[0083] (6) Add recovery and sample pretreatment:

[0084] Rice was selected as the test sample. The sample was pulverized and passed through a 20-mesh standard sieve. Three 20g portions of the sample were weighed, and 0.5 ppb, 2 ppb, and 5 ppb of terfenadine standard were added to each portion respectively (based on the antibody linear range and IC50). 50 (Set the concentration), add 10 mL of water, vortex mix, let stand for 30 min, and filter. Add 50 mL of acetone to the sample, shake on an electric shaker for 30 min, filter through rapid qualitative filter paper into a beaker, extract the residue again with 30 mL of acetone using the above method, wash the residue twice with 30 mL of acetone, combine the washings in a beaker, concentrate to near dryness in a 50℃ water bath, and reconstitute with 5 mL of 10% acetone-PBS solution (i.e., diluted five times to reduce the influence of the sample matrix).

[0085] The recovery rates of telexambin were 89.34-112.4% using indirect competitive ELISA.

[0086] 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 that secretes telexambin monoclonal antibody, characterized in that: The hybridoma cell line has the accession number CGMCC NO.46230.

2. The use of the hybridoma cell line according to claim 1 in the preparation of telexambin monoclonal antibody.

3. A monoclonal antibody against telexambin, characterized in that: The telexin monoclonal antibody is secreted by the hybridoma cell line described in claim 1.

4. A composition, characterized in that: The composition comprises the monoclonal antibody of claim 3.

5. The use of the telexin monoclonal antibody of claim 3 or the composition of claim 4 in the preparation of telexin detection products.

6. The application according to claim 5, characterized in that: The telexin detection products include reagents, test strips, kits, or biochips.

7. A test strip, characterized in that: The test strip contains the telexin monoclonal antibody as described in claim 3 or the composition as described in claim 4.

8. A reagent kit, characterized in that: The kit contains the telexin monoclonal antibody of claim 3 or the composition of claim 4.

9. The reagent kit according to claim 8, characterized in that: The kit also includes an ELISA plate, telexin-coated antigen, telexin standard solution, enzyme-labeled secondary antibody, and colorimetric solution.

10. The use of the telexin monoclonal antibody of claim 3, the composition of claim 4, the test strip of claim 7, or the kit of claim 8 in the detection of telexin.

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