A pentachlorophenol sodium hapten, artificial antigen and their application in immunoassay

Abstract

The present invention discloses a pentachlorophenol sodium hapten, an artificial antigen thereof and their applications in immunoassay. The structure of the pentachlorophenol sodium hapten is shown in Formula I, and the structural formula of the pentachlorophenol sodium artificial antigen is shown in Formula II. The preparation method of the pentachlorophenol sodium hapten of the present invention is simple. The introduced arm of the designed pentachlorophenol sodium hapten not only has active groups, but also completely retains the phenol structure of the target analyte, increasing the recognition sites. The content of the product pentachlorophenol sodium hapten has been greatly improved. Moreover, the pentachlorophenol sodium artificial antigen and monoclonal antibody prepared based on the hapten of the present invention have strong detection specificity. When used in test strips, they can detect more trace amounts of pentachlorophenol sodium, with a detection sensitivity of 0.8 μg / L for pentachlorophenol sodium and higher sensitivity.

Description

Technical Field

[0001] The present invention relates to the field of biochemistry, and particularly to a pentachlorophenol sodium hapten, an artificial antigen and their applications in immunoassay. Background Art

[0002] Sodium pentachlorophenoxide (PCP-Na) is a kind of compound with the structural feature of pentachlorophenoxy group, and is widely used as a fungicide, herbicide, insecticide and wood preservative. Sodium pentachlorophenoxide has stable chemical properties and is not easily degraded, and can enter the food chain through bioaccumulation. Sodium pentachlorophenoxide has teratogenic, carcinogenic and mutagenic effects. After long-term ingestion of sodium pentachlorophenoxide by humans, it will damage the human immune function and nerve function, and has great harmfulness. Worldwide, most countries, regions and organizations have established relevant laws and regulations, listing sodium pentachlorophenoxide in the veterinary drugs prohibited for food animals, prohibiting its use in aquaculture of aquatic animals, and it shall not be detected in animal-derived foods.

[0003] At present, the conventional instrumental detection method not only has expensive equipment and long detection time, but also requires professional operation, and cannot realize on-site rapid spot check detection; another detection method is immunoassay technology, which has the characteristics of high specificity and high selectivity and is very suitable for the detection of trace components in complex matrices. The performance of immunoassay products is determined by the performance of antigens and antibodies, and the key to antigens and antibodies is haptens. Therefore, to obtain excellent antigens and antibodies, the structural design of haptens is particularly important. Therefore, the development of highly specific pentachlorophenol sodium haptens or artificial antigens is of crucial significance for rapid, highly sensitive and low-cost detection methods of pentachlorophenol sodium.

[0004] CN113754532A discloses a pentachlorophenol sodium hapten, and its structural formula is . The IC 50 value of the artificial antigen and antibody prepared using the pentachlorophenol sodium hapten in the invention can reach 1.07 μg / L, and the actual detection sensitivity for pentachlorophenol sodium is at least 1 μg / L.

[0005] CN105116148A discloses a test strip and method for detecting pentachlorophenol sodium. The test line of the test strip is coated with a pentachlorophenol sodium hapten-carrier protein conjugate, and the structural formula of the pentachlorophenol sodium hapten used therein is . The detection sensitivity of the test strip for pentachlorophenol sodium is 1 μg / L. The sensitivity of the existing pentachlorophenol sodium haptens needs to be further improved. Summary of the Invention

[0006] The object of the present invention is to overcome at least one deficiency of the prior art and provide a pentachlorophenol sodium hapten, an artificial antigen thereof and an application thereof in immunoassay.

[0007] The technical solution adopted by the present invention is as follows:

[0008] In the first aspect, the present invention provides a pentachlorophenol sodium hapten, and the hapten has a structure shown in Formula I: Formula I.

[0009] In the second aspect, the preparation method of the pentachlorophenol sodium hapten provided by the present invention includes the following steps: Compound 1 and Compound 2 undergo a substitution reaction to obtain Compound 3; then Compound 3 is hydrolyzed to obtain the pentachlorophenol sodium hapten; wherein, the structural formula of Compound 1 is ; the structural formula of Compound 2 is ; the structural formula of Compound 3 is .

[0010] In some examples, the molar dosage of Compound 1 is more than 5 times that of Compound 2, and the reaction temperature of Compound 1 and Compound 2 is 20 - 30 °C.

[0011] In the third aspect, the present invention provides a pentachlorophenol sodium artificial antigen, and the pentachlorophenol sodium artificial antigen is obtained by coupling the pentachlorophenol sodium hapten with a protein carrier.

[0012] In some examples, the structural formula of the pentachlorophenol sodium artificial antigen described in the third aspect of the present invention is shown in Formula II: Formula II; wherein, protein is a protein carrier.

[0013] In some examples, the protein carrier is selected from at least one of bovine serum albumin, ovalbumin, human serum albumin or hemocyanin.

[0014] In the fourth aspect, the preparation method of the pentachlorophenol sodium artificial antigen provided by the present invention includes the following steps: Mix the hapten shown in Formula I, carbodiimide and N-hydroxysuccinimide, and carry out an activation reaction to obtain a hapten activated ester; then mix the carrier protein with the hapten activated ester, and carry out an amidation reaction to obtain the pentachlorophenol sodium artificial antigen.

[0015] In some examples, the activation reaction time is 3 - 5 h, and the amidation reaction time is 16 - 24 h.

[0016] In the fifth aspect, the present invention provides an application of the pentachlorophenol sodium artificial antigen in a pentachlorophenol sodium ELISA detection method.

[0017] Sixthly, the present invention provides a test strip or test reagent for detecting sodium pentachlorophenol, and the test strip or test reagent contains the artificial antigen of sodium pentachlorophenol.

[0018] The beneficial effects of the present invention are as follows:

[0019] 1. The arm introduced in the hapten of sodium pentachlorophenol designed by the present invention not only has active groups, but also completely retains the phenol structure of the target analyte, enhancing the recognition site. Due to the large number of recognition sites in the present invention, the content of the product hapten of sodium pentachlorophenol has been greatly improved;

[0020] 2. The artificial antigen and monoclonal antibody of sodium pentachlorophenol in the present invention are highly specific for ELISA detection, and the IC 50 value is 0.87 μg / L, which is superior to the existing antigens;

[0021] 3. The artificial antigen and monoclonal antibody of sodium pentachlorophenol in the present invention are used in the colloidal gold immunochromatography technology, which can quickly and conveniently detect sodium pentachlorophenol. Moreover, the colloidal gold immunochromatography test strip prepared in the present invention has a detection sensitivity of 0.8 μg / L for sodium pentachlorophenol, and can detect more trace sodium pentachlorophenol, with higher sensitivity. Brief Description of the Drawings

[0022] Figure 1 is the synthesis route diagram of the hapten of sodium pentachlorophenol;

[0023] Figure 2 is the mass spectrum diagram of the hapten of sodium pentachlorophenol prepared by the present invention;

[0024] Figure 3 is the synthesis route diagram of the artificial antigen of sodium pentachlorophenol;

[0025] Figure 4 is the standard curve diagram of sodium pentachlorophenol in the ELISA test. Detailed Embodiments

[0026] The following disclosure provides many different embodiments or examples for implementing different solutions of the present invention.

[0027] Example 1 (Preparation of the Hapten of Sodium Pentachlorophenol)

[0028] As Figure 1 shown, the hapten of sodium pentachlorophenol in this example is prepared by the following method, and the steps are as follows:

[0029] 1) Weigh 1.00 g (4.00 mmol) of Compound 1 into a 100-ml round-bottom flask. Then, successively add 10 ml of DMF, 0.55 g of K2CO3 (4.00 mmol), 0.16 g (0.80 mmol) of Compound 2, and a catalytic amount of tetrabutylammonium bromide (50 mg). React at 25 °C for more than 16 h. After the reaction is completed, add 90 ml of purified water, extract twice with ethyl acetate, then remove the solvent under reduced pressure, and purify by column chromatography to obtain 0.26 g of Compound 3.

[0030] 2) Dissolve 0.26 g (0.72 mmol) of Compound 3 in 5 ml of methanol, then add it to 5 ml of 4 mol / L lithium hydroxide aqueous solution. The reaction mixture reacts at RT for more than 8 h. Add 30 ml of sodium chloride aqueous solution, and extract twice with 30 ml of dichloromethane. Adjust the pH value of the aqueous phase to 3 - 4 with 4 M dilute hydrochloric acid, extract with ethyl acetate, then remove the solvent under reduced pressure, and purify by column chromatography to obtain 0.11 g of hapten. The mass spectrum is as Figure 2 shown.

[0031] Comparative Example 1 (Preparation of pentachlorophenol sodium hapten)

[0032] As Figure 1 shown, the pentachlorophenol sodium hapten of this example is prepared by the following method. The steps are as follows:

[0033] 1) Weigh 0.50 g (2.00 mmol) of Compound 1 into a 100-ml round-bottom flask. Then, successively add 10 ml of DMF, 0.55 g of K2CO3 (4.00 mmol), 0.16 g (0.80 mmol) of Compound 2, and a catalytic amount of tetrabutylammonium bromide (50 mg). React at 25 °C for more than 16 h. After the reaction is completed, add 90 ml of purified water, extract twice with ethyl acetate, then remove the solvent under reduced pressure, and purify by column chromatography to obtain 0.23 g of Compound 3.

[0034] 2) Dissolve 0.233 g (0.645 mmol) of Compound 3 in 5 ml of methanol, then add it to 5 ml of 4 mol / L lithium hydroxide aqueous solution. The reaction mixture reacts at RT for more than 8 h. Add 30 ml of sodium chloride aqueous solution, and extract twice with 30 ml of dichloromethane. Adjust the pH value of the aqueous phase to 3 - 4 with 4 M dilute hydrochloric acid, extract with ethyl acetate, then remove the solvent under reduced pressure, and purify by column chromatography to obtain 0.10 g of hapten.

[0035] Comparative Example 2 (Preparation of pentachlorophenol sodium hapten)

[0036] As Figure 1 shown, the pentachlorophenol sodium hapten of this example is prepared by the following method. The steps are as follows:

[0037] 1) Take 1.00 g (4.00 mmol) of Compound 1 in a 100 ml round-bottom flask, and then successively add 10 ml of DMF, 0.55 g of K2CO3 (4.00 mmol), 0.16 g (0.80 mmol) of Compound 2 and a catalytic amount of tetrabutylammonium bromide (50 mg). React at 35 °C for more than 16 h. After the reaction is completed, add 90 ml of purified water, extract twice with ethyl acetate, then remove the solvent under reduced pressure, and purify by column chromatography to obtain 0.26 g of Compound 3.

[0038] 2) Dissolve 0.225 g (0.623 mmol) of Compound 3 in 5 ml of methanol, then add it to 5 ml of 4 mol / L lithium hydroxide aqueous solution. The reaction mixture reacts at RT for more than 8 h. Add 30 ml of sodium chloride aqueous solution, and extract twice with 30 ml of dichloromethane. Adjust the pH value of the aqueous phase to 3 - 4 with 4M dilute hydrochloric acid, extract with ethyl acetate, then remove the solvent under reduced pressure, and purify by column chromatography to obtain 0.095 g of hapten.

[0039] Synthesis of Sodium Pentachlorophenol Artificial Antigen with Bovine Serum Albumin as Carrier Protein

[0040] As Figure 3 shown, the sodium pentachlorophenol artificial antigen with bovine serum albumin as carrier protein is synthesized as follows:

[0041] 1) Take 10 mg of the sodium pentachlorophenol hapten prepared in Example 1 and dissolve it in 0.2 mL of dimethylformamide (DMF). After stirring well, add 10 mg of carbodiimide (EDC) and 10 mg of N-hydroxysuccinimide (NHS), and stir at room temperature for 4 h to obtain the hapten activated ester.

[0042] 2) Weigh 30 mg of bovine serum albumin (BSA) and dissolve it thoroughly in 3 mL of 0.01 mol / L PBS solution to form a carrier protein solution. While stirring, slowly add the hapten activated ester drop by drop to the carrier protein solution, and stir at room temperature for 16 - 24 h.

[0043] 3) Dialyze the solution prepared in step 2) with 0.01 mol / L PBS at room temperature for 3 days, changing the dialysis solution 3 times a day to remove unreacted small molecule substances.

[0044] 4) Aliquot and store at 4 °C for later use.

[0045] Synthesis of Sodium Pentachlorophenol Artificial Antigen with Hemocyanin as Carrier Protein

[0046] As Figure 3 shown, the sodium pentachlorophenol artificial antigen with hemocyanin as carrier protein is synthesized as follows:

[0047] 1) Take 5 mg of the sodium pentachlorophenol hapten prepared in Example 1, dissolve it in 0.2 mL of N,N-dimethylformamide (DMF), and after stirring thoroughly, add 10 mg of EDC and 10 mg of N-hydroxysuccinimide (NHS), and stir at room temperature for 4 h to obtain the activated ester of the hapten;

[0048] 2) Weigh 15 mg of keyhole limpet hemocyanin (KLH), dissolve it sufficiently in 3 mL of 0.01 mol / L PBS solution to form a carrier protein solution, and slowly dropwise add the activated ester of the hapten to the carrier protein solution drop by drop under stirring, and stir at room temperature for 16 - 24 h;

[0049] 3) Dialyze the solution prepared in step (2) at room temperature with 0.01 mol / L PBS for 3 days, changing the dialysis solution 3 times a day to remove unreacted small molecules;

[0050] 4) Aliquot and store for later use at 4°C.

[0051] Application of the sodium pentachlorophenol artificial antigen in the preparation of monoclonal antibodies against sodium pentachlorophenol

[0052] The method for preparing monoclonal antibodies against sodium pentachlorophenol is as follows:

[0053] Using the sodium pentachlorophenol artificial antigen with the above carrier protein being keyhole limpet hemocyanin as the immunogen, emulsify it with an equal volume of Freund's adjuvant and immunize BALB / C mice. The immunization dose for each mouse is 100 μg, the immunization interval is 2 weeks, and after immunizing 3 times, collect the blood from the tail vein of the mice to detect the serum titer. If the antibody titer does not meet the requirements, booster immunization is required. After the antibody titer no longer increases, boost immunize subcutaneously with 100 μg of the whole antigen. 5 days later, take the spleen cells of the mice and fuse them with SP20 cells (mouse myeloma cells). The fused cells are screened in HAT medium, and after 5 days, replace the HAT medium with complete medium for culture. Detect the cell supernatant by ELISA, and clone and culture the cells in the wells with strongly positive detection results by the limiting dilution method. After 3 times of clone culture and detection, the cells in the wells that are all positive are the hybridoma cells secreting monoclonal antibodies. After amplifying and culturing the hybridoma cells, inoculate them into the abdominal cavity of mice to produce ascites containing antibodies. Purify the ascites by the caprylic acid-ammonium sulfate precipitation method, and high-purity and high-specificity monoclonal antibodies can be obtained after lyophilization.

[0054] Application and effect evaluation of the sodium pentachlorophenol artificial antigen in ELISA

[0055] Using carbonate buffer solution with pH 9.6 as the coating dilution solution, dilute the artificial antigen of sodium pentachlorophenol with bovine serum albumin as the carrier protein to 0.08 μg / mL, add 100 μL per well into the polystyrene microplate, coat overnight at 4°C, drain, add 280 μL per well of 1% BSA, and block at 37°C for 1 h in phosphate buffer solution, drain, and store in vacuum packaging after drying.

[0056] Add 100 μL per well of sodium pentachlorophenol standard solution (gradient diluted with phosphate buffer solution with pH 7.4) to the microplate enzyme-linked immunosorbent assay (ELISA) plate coated with the artificial antigen of sodium pentachlorophenol, and then add 20 μL per well of the monoclonal antibody solution of sodium pentachlorophenol (dilute the monoclonal antibody to 0.1 μg / mL with phosphate buffer solution containing 0.05% sodium azide and pH 7.4, freshly prepared), react at 37°C for 0.5 h; after draining, add 250 μL per well of the washing solution, wash 3 times and then pat dry; then add 100 μL per well of the enzyme-labeled secondary antibody, react at 37°C for 0.5 h; wash 3 times again and pat dry, add 100 μL per well of the chromogenic solution, react at 37°C for 15 min; add 50 μL per well of 0.5 M sulfuric acid to terminate, and set the ELISA reader to measure the OD value of each well at a wavelength of 450 nm. The results are shown in Table 1 below:

[0057] Table 1. OD values of ELISA testing standard solutions of sodium pentachlorophenol at different concentrations

[0058]

[0059] Based on the data in Table 1, use ELISA Calc software to perform four-parameter Logistic curve fitting to draw the standard curve. The obtained standard curve is as Figure 2 shown. The linear equation of sodium pentachlorophenol is: y = (A - D) / [1 + (X / C)^B] + D, r 2 = 0.998, A = 2.33658, B = 0.78860, C = 0.86561, D = 0.00171, x represents the concentration of the analyte to be measured, y represents the OD value, and the IC 50 value is calculated to be 0.87 μg / L, showing a linear relationship in the range of 0.2 - 16.2 μg / L.

[0060] Preparation of colloidal gold qualitative immunochromatographic reagent strip for sodium pentachlorophenol

[0061] The preparation method of the colloidal gold qualitative immunochromatographic reagent strip for sodium pentachlorophenol is as follows:

[0062] 1. Prepare the reaction membrane coated with artificial antigen and goat anti-mouse IgG:

[0063] Using a nitrocellulose membrane (NC membrane) as the reaction membrane, adjust the concentration of the artificial antigen with bovine serum albumin as the carrier protein to 0.1 - 0.2 mg / mL with coating buffer, and also adjust the concentration of goat anti-mouse IgG to 0.1 - 1 mg / mL with coating buffer. Spray the antigen and goat anti-mouse IgG onto the corresponding test area (T line) and control area (C line) of the reaction membrane according to a membrane liquid volume of 0.8 - 1.2 μL / cm. The interval between the test area and the control area is 2.5 mm. Place it in an oven at 37 °C for 16 - 24 h, and store it in a constant temperature and humidity storage box for later use; the coating buffer is 0.01 M PBS buffer with a pH of 7.4 containing 2% sucrose and 0.05% sodium azide.

[0064] 2. Preparation of the gold pad with nano-gold labeled sodium pentachlorophenol monoclonal antibody:

[0065] a) Preparation of nano-gold:

[0066] Preparation of nano-gold solution: Take 1 g of chloroauric acid, dissolve it with pure water and ultrasonic waves, and make up the volume to 100 ml. Store it in the dark at 4 °C for later use. Take 1 ml of the above solution into 100 ml of pure water, heat it to boiling, then add 0.5 ml of 0.06% sodium citrate solution, continue heating for 10 minutes, cool it to room temperature, and restore the volume to the original volume (100 ml) with pure water. Store it in the dark at room temperature for later use. All glassware used needs to be soaked overnight in a mixed solution of potassium permanganate and sulfuric acid, and used after cleaning and drying.

[0067] b) Labeling of sodium pentachlorophenol monoclonal antibody:

[0068] Add 6 μg of sodium pentachlorophenol monoclonal antibody to 1 ml of 0.01% nano-gold solution, react at room temperature for 10 minutes, add 10 μl of 10% bovine serum albumin for blocking, and centrifuge at 10000 rpm to discard all the supernatant.

[0069] c) Preparation of the gold pad:

[0070] Redissolve with 1 ml of gold dilution solution containing 2% Tris, 5% bovine serum albumin, 0.05% thimerosal, and 5% sucrose, and evenly smear it on a glass fiber with an area of 5 cm * 5 cm. Dry it at 37 °C for 16 - 24 h and store it for later use.

[0071] 3. Preparation of the sample pad:

[0072] Soak the cut blank sample pad of 30 * 30 cm in the sample pad treatment solution for 5 minutes, take it out and dry it at 37 °C for 16 - 24 h, and store it in a constant temperature and humidity storage box for later use; the sample pad treatment solution is 0.1 M PB buffer containing 0.3% Tween 20, 1% sucrose, 0.5% BSA, and 0.05% sodium azide.

[0073] 4. Assembly of colloidal gold qualitative immunochromatographic test strip:

[0074] Stack the reaction membrane prepared in step (1) in the middle of the PVC board backing, stack the gold pad and the absorbent pad of step (2) at both ends respectively. The reaction membrane is connected to the gold pad and the absorbent pad, the gold pad is connected to the sample pad, the detection area is close to the sample pad, and the control area is close to the absorbent pad to obtain a test paper board. Cut the test paper board into 3-mm test strips, load the test strips into a test paper card to obtain a colloidal gold qualitative immunochromatographic test paper card, and then further test the sodium pentachlorophenol colloidal gold immunochromatographic test paper card prepared by the present invention.

[0075] Detection limit test of sodium pentachlorophenol colloidal gold immunochromatographic test paper card

[0076] Prepare a series of standard solutions of sodium pentachlorophenol with different concentrations using 0.01M PBS buffer solution, and then take 100 μl of the standard solution and add it to the sample adding hole of the sodium pentachlorophenol colloidal gold immunochromatographic test paper card of the present invention. Start timing after adding the sample. The result can be observed in 5 - 8 minutes, and the reading is invalid after 8 minutes.

[0077] Naked-eye reading method: If the color development of the T line is stronger than that of the C line or there is no obvious difference from the color development of the C line, it indicates that the test result is negative (-). If the color development of the T line is significantly weaker than that of the C line or the T line does not develop color, it indicates that the test result is positive (+). Invalid: If the C line does not appear, it indicates that the operation is incorrect or the test strip has failed. Set 3 groups of repetitions for the detection test. The rapid qualitative detection of the colloidal gold immunochromatographic test paper card can be realized.

[0078] The measurement results are as follows:

[0079] Table 2 Measurement results of colloidal gold qualitative immunochromatographic test paper card for detecting standard solutions of sodium pentachlorophenol with different concentrations

[0080] Standard solution concentration (μg / L) 0.1 0.2 0.4 0.8 1.6 3.2 6.4 Sodium pentachlorophenate — — — + + + +

[0081] By detecting the concentration values of the sodium pentachlorophenol series standard solutions, the results are shown in Table 2. The test strips of the present invention can detect sodium pentachlorophenol with different concentrations (Table 2). Therefore, the colloidal gold qualitative immunochromatographic test strips prepared by the present invention have high sensitivity for the detection of sodium pentachlorophenol.

[0082] Stability test of colloidal gold qualitative immunochromatographic test paper card

[0083] The storage condition of the colloidal gold qualitative immunochromatographic test strip is room temperature. To ensure the stability of the test strip, an accelerated destructive experiment was conducted on the test strip. It was continuously placed at 45°C for 60 days, and the color change of the negative and sulfamethazine standard solution was detected on the 0th day, 5th day, 10th day, 20th day, 30th day, 40th day, 50th day, and 60th day respectively. The experiment was set with 3 replicates, and the results are shown in Table 3: ("+" represents positive, "-" represents negative).

[0084] Table 3 Stability test

[0085]

[0086] As can be seen from Table 3, after 60 days, the T / C color development depth reading results of the colloidal gold qualitative immunochromatographic test strip stored sealed at 45°C showed no obvious change, indicating that the colloidal gold qualitative immunochromatographic test strip can be stably stored for at least 60 days at 45°C in the accelerated experiment. Therefore, the sodium pentachlorophenol colloidal gold qualitative immunochromatographic test strip prepared by the present invention can be stably stored at room temperature for more than one year, fully meeting the requirements of the market during storage and transportation.

[0087] The above is a further detailed description of the present invention and should not be regarded as a limitation on the specific implementation of the present invention. For those of ordinary skill in the technical field to which the present invention pertains, any simple deduction or substitution without departing from the concept of the present invention falls within the protection scope of the present invention.

Claims

1. A pentachlorophenol sodium hapten, characterized in that, The hapten has the structure shown in Formula I: Formula I.

2. The preparation method of the sodium pentachlorophenol hapten according to claim 1, characterized in that, It includes the following steps: 1) Compound 1 and Compound 2 undergo a substitution reaction to obtain Compound 3; 2) Compound 3 is hydrolyzed to obtain the sodium pentachlorophenol hapten; Among them, the structural formula of Compound 1 is ; the structural formula of Compound 2 is ; The structural formula of Compound 3 is .

3. The preparation method according to claim 2, wherein The molar dosage of Compound 1 is more than 5 times that of Compound 2, and the temperature for the substitution reaction of Compound 1 and Compound 2 is 20 - 30 °C.

4. An artificial antigen of sodium pentachlorophenol, characterized in that, The sodium pentachlorophenol artificial antigen is obtained by coupling the sodium pentachlorophenol hapten described in Claim 1 with a protein carrier.

5. The sodium pentachlorophenol artificial antigen according to claim 4, wherein, The structural formula of the sodium pentachlorophenol artificial antigen is shown in Formula II: Formula II; wherein, protein is a protein carrier.

6. The sodium pentachlorophenolate artificial antigen according to claim 4 or 5, characterized in that, The protein carrier is selected from at least one of bovine serum albumin, ovalbumin, human serum albumin, or hemocyanin.

7. The preparation method of the sodium pentachlorophenol artificial antigen according to any one of claims 4 to 6, characterized in that, It includes the following steps: 1) The hapten shown in Formula I, carbodiimide, and N-hydroxysuccinimide are mixed, and an activation reaction is carried out to obtain the hapten activated ester; 2) The carrier protein and the hapten activated ester are mixed, and an amidation reaction is carried out to obtain the sodium pentachlorophenol artificial antigen.

8. The preparation method according to claim 7, characterized in that, The activation reaction time is 3 - 5 h, and the amidation reaction time is 16 - 24 h.

9. Use of the sodium pentachlorophenol artificial antigen according to any one of Claims 4 - 6 in a sodium pentachlorophenol ELISA detection method.

10. A test strip or test reagent for detecting sodium pentachlorophenate, characterized in that, The test strip or test reagent contains the sodium pentachlorophenol artificial antigen according to any one of Claims 4 - 6.

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