CN113637642B - Hybridoma cell strain secreting chlorfenapyr monoclonal antibody and application thereof - Google Patents
Hybridoma cell strain secreting chlorfenapyr monoclonal antibody and application thereof Download PDFInfo
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Abstract
The invention discloses a hybridoma cell strain secreting a chlorfenapyr monoclonal antibody and application thereof, and belongs to the technical field of immunodetection. The invention discloses a hybridoma cell strain JHJ secreting a chlorfenapyr monoclonal antibody, which is preserved in China general microbiological culture Collection center (CGMCC), and is classified and named as the monoclonal cell strain JHJ, the preservation date is 2021, the month is 13, and the preservation number is CGMCC No.22329. According to the invention, the BALB/c mice are immunized by using the trichloro acaricidal hol complete antigen, spleen cells of the mice with high titer and excellent inhibition are taken and fused with myeloma cells by a PEG method, and hybridoma cell strains are obtained through screening and three subcloning by an indirect competitive ELISA method. Monoclonal antibodies secreted by this cell lineHas better specificity and detection sensitivity (IC) to the chlorfenapyr 50 The value is 2 ng/mL), provides raw materials for the immunodetection of the residual chlorfenapyr in food, and has practical application value.
Description
Technical Field
The invention belongs to the technical field of immunodetection, and particularly relates to a hybridoma cell strain secreting a chlorfenapyr monoclonal antibody and application thereof.
Background
Trichlorfon (dicofos), also known as 1, 1-di (p-chlorophenyl) -2, 2-trichloroethanol or kele powder, is an organic chlor-acaricide commonly used in agricultural production, and is often used for preventing and controlling various mites of cotton, fruit trees and flowers in China. However, in the process of producing the trichlorfon, a certain amount of DDT residues may be contained, which is harmful to the environment and human health. There has been increasing evidence in recent years that exposure to the environment is toxic to fish, reptiles, birds, mammals and humans and estrogenic effects, with extremely high toxicity to aquatic organisms. Therefore, the application of the trichlorfon in agricultural production is forbidden in China. 10.27 in 2017, the world health organization international cancer research institutes published a preliminary collation reference for the list of carcinogens, and the trichlorfon is in the list of 3 types of carcinogens. The maximum residual limit of pesticide in food GB 2763-2021 specifies the maximum residual limit standard (MRL) of the trichlorfon in various foods, such as 0.01mg/kg for vegetables and fruits, 0.02mg/kg for nuts and 0.02mg/kg for rice, wheat and miscellaneous cereals.
In order to effectively monitor the use condition of the trichlorfon in food, a measuring method with good specificity and high sensitivity is needed, but the existing detecting method such as gas chromatography, high performance liquid chromatography tandem mass spectrometry and the like has the defects of high cost, complicated sample pretreatment, high technical requirements for operators, incapability of immediately outputting results and more interferents in the food, so that the instrument method is not suitable for on-site detection. The immunological rapid detection method is a low-cost, rapid, high-sensitivity and high-specificity immunological detection method, is simple and convenient to operate, and is widely applicable to the field of pesticide residue detection. An efficient immunological detection method is established, and the screening of monoclonal antibodies with high specificity is an important precondition.
Disclosure of Invention
The invention aims to: in order to overcome the defects in the prior art, the invention provides a hybridoma cell strain secreting the monoclonal antibody of the chlorfenapyr and application thereof, and the antibody prepared by the cell strain has higher detection sensitivity to the chlorfenapyr and can be used for establishing an immunological detection method of the chlorfenapyr.
The technical scheme is as follows: in order to achieve the above purpose, the invention adopts the following technical scheme:
the first object of the present invention is to provide a hybridoma cell strain JHJ secreting the monoclonal antibody of the chlorfenapyr, which is preserved in China general microbiological culture Collection center (CGMCC), the institute of microbiological research, national academy of sciences of China, no. 3, north Chen West Lu 1, korea, beijing, the collection date 2021, 05 month 13, and the preservation number CGMCC No.22329.
The monoclonal antibody of the trichlorethamate is secreted by a hybridoma cell strain JHJ of the monoclonal antibody of the trichlorethamate with the preservation number of CGMCC No.22329.
The second object of the invention is to provide a trichlorethamine hapten, which has the following structural formula:
the third object of the invention is to provide a method for synthesizing a trichlorfon hapten, which comprises the following steps: adding 4,4' -dichloro diphenyl ketone and carboxymethyl hydroxylamine semi-hydrochloride solid into a mixed solution composed of methanol, pyridine and pure water, and continuously stirring in a water bath at 60-90 ℃ for reaction for 5-10 h; taking out, and standing overnight at room temperature; drying with nitrogen, adding dichloromethane for redissolution, extracting with pure water, discarding the water layer, and drying the organic phase with nitrogen to obtain a viscous solid which is the chlorfenapyr hapten.
In one embodiment of the present invention, the molar mass ratio of the 4,4' -dichlorobenzophenone to the carboxymethyl hydroxylamine hemi-hydrochloride solid is 1: 3-6, wherein the concentration of the 4,4' -dichlorobenzophenone in the mixed solution is 5-10 g/L; preferably, the molar mass ratio of the 4,4' -dichlorobenzophenone to the carboxymethyl hydroxylamine half hydrochloride solid is 1:4, the concentration of the 4,4' -dichlorobenzophenone in the mixed solution is 8.3g/L.
In one embodiment of the invention, the mixed solution is formed by methanol, pyridine and pure water according to a volume ratio of 2-5:1-3:1; preferably, the mixed solution is formed by methanol, pyridine and pure water according to a volume ratio of 4:1:1.
The fourth object of the invention is to provide a method for preparing a complete antigen of the trichlorfon, wherein the complete antigen comprises an immunogen of the trichlorfon, and the method comprises the following steps: dissolving the chlorfenapyr hapten with N, N-dimethylformamide, adding 1-ethylcarbodiimide hydrochloride and N-hydroxysuccinimide, and continuously stirring at room temperature for reaction to obtain a chlorfenapyr hapten activating solution; dissolving carrier protein with boric acid buffer solution, which is called protein A solution; slowly dripping the trichlorfon hapten activating solution into the protein A solution at room temperature, stirring at room temperature for reaction overnight, and dialyzing to obtain the trichlorfon complete antigen.
In one embodiment of the invention, the carrier proteins include keyhole limpet hemocyanin, bovine serum albumin, BSA, and other carrier proteins suitable for coupling.
In one embodiment of the invention, the mole mass ratio of the trichlorfon hapten, the 1-ethylcarbodiimide hydrochloride and the N-hydroxysuccinimide is 1:3 to 5: 3-5, wherein the concentration of the chlorfenapyr hapten in the N, N-dimethylformamide is 20-30 g/L; preferably, the mole mass ratio of the trichlorfon hapten, the 1-ethylcarbodiimide hydrochloride and the N-hydroxysuccinimide is 1:4.1:3.4, the concentration of the trichlorfon hapten in N, N-dimethylformamide is 25g/L.
In one embodiment of the invention, the concentration of the carrier protein in the boric acid buffer solution is 3-5 g/L; preferably, the carrier protein is present in the boric acid buffer at a concentration of 3.3g/L.
In one embodiment of the invention, the molar mass ratio of the trichlorfon hapten to the carrier protein is 6000-8000: 1, a step of; preferably, the molar mass ratio of the trichlorfon hapten to the carrier protein is 6160:1.
in one embodiment of the invention, the structural formula of the trichlorfon hapten is as follows:
the fifth object of the present invention is to provide a method for preparing a complete antigen of chlorfenapyr, wherein the complete antigen comprises a chlorfenapyr coating antigen, and the method comprises the following steps: dissolving 2, 2-bis (p-chlorophenyl) acetic acid with anhydrous N, N-dimethylformamide, adding 1-ethylcarbodiimide hydrochloride and N-hydroxysuccinimide, and continuously stirring at room temperature for reaction to obtain a trichlorfon hapten activating solution; the carrier protein is dissolved in a boric acid buffer solution, which is called solution B; and under the condition of room temperature, dropwise adding the trichloro acaricidal hapten activating solution into the solution B, stirring at room temperature for reaction overnight, and dialyzing to obtain the trichloro acaricidal coating antigen.
In one embodiment of the invention, the carrier protein comprises chicken ovalbumin OVA, bovine serum albumin BSA and other carrier proteins suitable for coupling.
In one embodiment of the present invention, the molar mass ratio of the 2, 2-bis-p-chlorophenyl acetic acid, 1-ethylcarbodiimide hydrochloride and N-hydroxysuccinimide is 1:3 to 5: 3-5, wherein the concentration of the 2, 2-bis-p-chlorophenyl acetic acid in the N, N-dimethylformamide is 10-25 g/L; preferably, the molar mass ratio of the 2, 2-bis-p-chlorophenyl acetic acid to the 1-ethylcarbodiimide hydrochloride to the N-hydroxysuccinimide is 1:3.5:3.0, wherein the concentration of the 2, 2-bis-p-chlorophenyl acetic acid in N, N-dimethylformamide is 20g/L.
In one embodiment of the invention, the concentration of the carrier protein in the boric acid buffer solution is 3-7 g/L; preferably, the carrier protein is present in the boric acid buffer at a concentration of 5g/L.
In one embodiment of the present invention, the molar mass ratio of the 2, 2-bis-p-chlorophenyl acetic acid to the carrier protein is 50 to 100:1, a step of; preferably, the molar mass ratio of the 2, 2-bis-p-chlorophenyl acetic acid to the carrier protein is 64:1.
in one embodiment of the invention, the preparation method of the hybridoma cell strain JHJ secreting the chlorfenapyr monoclonal antibody provided by the invention comprises the following basic steps:
(1) Synthesis of hapten:
the synthesis scheme of the trichlorfon hapten is shown in the reaction formula. 4,4' -dichlorobenzophenone (15 mg) and carboxymethyl hydroxylamine hemi-hydrochloride (30 mg) were weighed into a 10mL screw-type brown glass bottle, 1.8mL of a mixed solution of methanol, pyridine and pure water (v: v: v=4:1:1) was added, followed by placing in a water bath at 70℃and continuously stirring the reaction for 6 hours. After the reaction was completed, the mixture was taken out and allowed to stand at room temperature overnight. After the solution is dried by nitrogen, 2mL of dichloromethane is added for redissolution, pure water is used for repeated extraction for 3-4 times (1 mL/time), the water layer is discarded, and the viscous solid obtained by drying the organic phase by nitrogen is the trichlorfon hapten.
(2) Preparation of immunogens and coating precursors:
preparation of the immunogen: 5.0mg of the chlorfenapyr hapten is weighed, 200 mu L of anhydrous N, N-dimethylformamide is used for dissolution, then 12mg of 1-ethylcarbodiimide hydrochloride and 6.0mg of N-hydroxysuccinimide are added, the mixture is uniformly mixed, and the reaction is continued at room temperature for 6 hours (called solution A). Dissolving key hole hemocyanin 10mg with 3.0mL boric acid buffer solution (called protein A solution), slowly adding the solution A into the protein A solution at room temperature, stirring at room temperature for reaction overnight, and dialyzing to obtain immunogen;
preparation of coating raw material: 4.0mg of 2, 2-bis-p-chlorophenyl acetic acid was weighed, dissolved in 200. Mu.L of anhydrous N, N-dimethylformamide, followed by adding 9.6mg of 1-ethylcarbodiimide hydrochloride and 4.8mg of N-hydroxysuccinimide, and mixing well, and the reaction was continued with stirring at room temperature for 6 hours (referred to as solution B). Weighing 10mg of chicken ovalbumin OVA, dissolving in 2mL of boric acid buffer solution (called solution B), dropwise adding the solution B into the solution B at room temperature, stirring at room temperature for reaction overnight, and dialyzing to obtain the coating antigen.
(3) Immunization of mice: after mixing and emulsifying the total antigen of the trichlorfon and the equal volume of the complete Freund adjuvant, BALB/c mice are subcutaneously injected through the nape of the neck. Complete Freund's adjuvant was used for the first immunization, and incomplete Freund's adjuvant was used for the booster immunization. Each boost was 3 weeks apart. The final boost was performed by intraperitoneal injection with complete antigen (without adjuvant); serum titers and inhibition were detected by indirect competitive enzyme-linked immunosorbent assay (ic-ELISA).
(4) Cell fusion and cell strain establishment: the method comprises the steps of fusing mouse spleen cells and myeloma cells by a PEG method, selectively culturing by a HAT culture medium, detecting the positive and inhibition rate of cell holes by ic-ELISA, subcloning hybridoma cells with high positive and inhibition rate by a limiting dilution method, and finally screening to obtain a trichlorfon monoclonal antibody hybridoma cell strain JHJ.
(5) Identification of hybridoma cell line properties: sensitivity was determined by ic-ELISA.
High potency low IC 50 The spleen cells of the mice are fused with myeloma cells of the mice by a PEG method, and a hybridoma cell strain JHJ is obtained through screening and subcloning by an indirect competitive enzyme-linked immunosorbent assay.
The sixth object of the invention is to provide an application of the monoclonal antibody of the trichlorfon, which is used for analysis and detection of trichlorfon residues in food safety detection.
In one embodiment of the invention, the monoclonal antibody of the trichlorfon is used for preparing a detection main body for analyzing and detecting the residual quantity of the trichlorfon in food safety detection.
In one embodiment of the present invention, the detection body includes a reagent, a detection plate, and a kit.
The invention has the beneficial effects that: firstly, the monoclonal antibody secreted by the cell strain JHJ provided by the invention has better specificity and detection sensitivity (IC) to the chlorfenapyr 50 The value is 2 ng/mL), can realize the detection of the residual quantity of the trichlorfon in fruits, vegetables and grains, provides a raw material for the immunodetection of the residual quantity of the trichlorfon in foods, and has practical application value. Secondly, the invention provides a preparation method of a hybridoma cell strain for preparing the monoclonal antibody capable of secreting the trichlorfon, in particular to a novel hapten preparation method, which is prepared by adding 4,4' -dichlorobenzophenone and carboxymethyl hydroxylamine half hydrochloride solid into a mixed solution composed of methanol, pyridine and pure water for reaction, and has the advantages of simple and easily obtained raw materials and simple and easy operation of the preparation process.
Preservation of biological material samples: the monoclonal antibody hybridoma cell strain JHJ of the trichlorfon is preserved in China general microbiological culture Collection center (CGMCC) of China Committee for culture Collection of microorganisms, and the division name is monoclonal cell strain JHJ, the preservation date 2021, 05 month and 13 days, and the preservation number is CGMCC No.22329 of the institute of microorganisms of national academy of sciences No. 3, north Chenxi, korea, beijing city.
Drawings
FIG. 1 is a schematic diagram showing the results of immunoglobulin subtype identification of the JHJ monoclonal antibody of the present invention.
FIG. 2 is a standard curve of inhibition of the chlorfenapyr by the JHJ monoclonal antibody of the present invention.
Detailed Description
The invention will be further described with reference to the drawings and examples. The invention will be better understood from the following examples. However, those skilled in the art will readily appreciate that the specific material ratios, process conditions and results thereof described in the examples are only illustrative of the present invention and should not be construed as limiting the scope of the present invention.
Reagents used in the examples of the present invention:
solution preparation: carbonate Buffer (CBS): weighing Na 2 CO 3 1.59g，NaHCO 3 2.93g, respectively dissolving in a small amount of double distilled water, mixing, adding double distilled water to about 800mL, mixing, adjusting pH to 9.6, adding double distilled water to 1000mL, and storing at 4deg.C for use;
phosphate Buffer (PBS): 8.00g NaCl,0.2g KCl,0.2g KH 2 PO 4 ，2.9g Na 2 HPO 4 ·12H 2 O is dissolved in 800mL of pure water, pH is regulated to 7.2-7.4 by NaOH or HCl, and volume is regulated to 1000mL;
PBST: PBS containing 0.05% Tween 20;
TMB color development liquid: and (3) solution A: na (Na) 2 HPO 4 . 12H 2 18.43g of O, 9.33g of citric acid and pure water to 1000mL; and (2) liquid B: 60mg of TMB was dissolved in 100mL of ethylene glycol. A. And mixing the solution B according to the volume ratio of 1:5 to obtain TMB color development solution, wherein the solution B is mixed immediately before use.
According to the embodiment of the invention, the mouse is immunized by the total antigen of the chlorfenapyr, the cell fusion is carried out, the HAT selective culture medium is used for culturing, and the cell supernatant is screened by the ic-ELISA, so that the monoclonal antibody hybridoma cell strain with higher sensitivity to the chlorfenapyr is finally obtained.
Example 1 preparation of hybridoma cell line JHJ
(1) Synthesis of hapten:
the synthesis scheme of the trichlorfon hapten is shown in the reaction formula. 4,4' -dichlorobenzophenone (15 mg,0.0597 mmol) and carboxymethyl hydroxylamine hemi-hydrochloride (30 mg,0.235 mmol) were weighed as solids into a 10mL screw-on brown glass bottle, 1.8mL of a mixed solution of methanol, pyridine and pure water (v: v=4:1:1) was added, followed by placing in a 70℃water bath with continuous stirring for reaction for 6h. After the reaction was completed, the mixture was taken out and allowed to stand at room temperature overnight. After the solution is dried by nitrogen, 2mL of dichloromethane is added for redissolution, pure water is used for repeated extraction for 3-4 times (1 mL/time), the water layer is discarded, and the viscous solid obtained by drying the organic phase by nitrogen is the trichlorfon hapten.
(2) Preparation of immunogens and coating precursors:
preparation of the immunogen: 5.0mg (0.0154 mmol) of the chlorfenapyr hapten is weighed, 200 mu L of anhydrous N, N-dimethylformamide is used for dissolution, then 12mg (0.0628 mmol) of 1-ethylcarbodiimide hydrochloride and 6.0mg (0.0522 mmol) of N-hydroxysuccinimide are added, and the mixture is uniformly mixed and stirred at room temperature for reaction for 6 hours (called solution A). Dissolving key hole hemocyanin 10mg (0.0000025 mmol) with 3.0mL boric acid buffer solution (called protein A solution), slowly adding the solution A into the protein A solution at room temperature, stirring at room temperature for reaction overnight, and dialyzing to obtain immunogen;
preparation of coating raw material: 4.0mg (0.0142 mmol) of 2, 2-bis-p-chlorophenyl acetic acid was weighed, dissolved with 200. Mu.L of anhydrous N, N-dimethylformamide, then 9.6mg (0.0503 mmol) of 1-ethylcarbodiimide hydrochloride, 4.8mg (0.0417 mmol) of N-hydroxysuccinimide was added, and the mixture was stirred uniformly at room temperature for 6 hours (referred to as solution B). 10mg (0.000222 mmol) of chicken ovalbumin OVA is weighed, dissolved in 2mL of boric acid buffer solution (called B solution), and the B solution is added dropwise into the B solution at room temperature, stirred at room temperature for reaction overnight and dialyzed to obtain the coating antigen.
(3) Immunization of mice: after mixing and emulsifying the total antigen of the trichlorfon and the equal volume of the complete Freund adjuvant, BALB/c mice are subcutaneously injected through the nape of the neck. Complete Freund's adjuvant was used for the first immunization, and incomplete Freund's adjuvant was used for the booster immunization. Each boost was 3 weeks apart. The final boost was performed by intraperitoneal injection with complete antigen (without adjuvant); serum titers and inhibition were detected by indirect competitive enzyme-linked immunosorbent assay (ic-ELISA).
(4) Cell fusion: three days after the last boost, cell fusion was performed according to the conventional PEG method, specifically as follows:
a. taking eyeball and blood, killing a mouse by a cervical dislocation method, immediately putting the mouse into 75% alcohol for disinfection, soaking for about 5min, taking out spleen of the mouse by aseptic operation, moderately grinding the spleen by a rubber head of a syringe, obtaining spleen cell suspension by a 200-mesh cell screen, collecting, centrifuging (1200 rpm,8 min), washing the spleen cells for three times by using an RPMI-1640 culture medium, and diluting the spleen cells to a certain volume and counting for later use after the last centrifugation;
b. collecting SP2/0 cells: SP2/0 tumor cells were cultured in 10% FBS (fetal bovine serum) RPMI-1640 medium at 5% CO 7-10 days prior to fusion 2 In an incubator. The number of SP2/0 tumor cells required before fusion reaches (1-4) x 10 7 Ensuring SP2/0 tumor cells to be in logarithmic growth phase before fusion. During fusion, collecting tumor cells, suspending in RPMI-1640 basic culture solution, and performing cell count;
c. the fusion process was 7min. 1min, 1mL of PEG 1500 was added dropwise to the cells from slow to fast; and (2) standing for 2 min. Dripping 1mL of RPMI-1640 culture medium in the period of 1min for 3min and 4 min; dripping 2mL of RPMI-1640 culture medium in the period of 1min at the 5 th and 6 th min; at 7min, 1mL of RPMI-1640 medium was added dropwise every 10 s. Then, the mixture was incubated at 37℃for 5min. Centrifuging (800 rpm,8 min), discarding supernatant, re-suspending in RPMI-1640 screening medium containing 20% fetal calf serum and 2% 50 XHAT, adding to 96-well cell plate at 200 μl/well, and standing at 37deg.C under 5% CO 2 Culturing in an incubator.
(5) Cell screening and cell strain establishment: the cells were subjected to half-replacement of the RPMI-1640 selection medium on day 3 of cell fusion, full replacement with a 100 XHT RPMI-1640 transition medium containing 20% fetal bovine serum and 1% on day 5, and cell supernatants were collected on day 7 for selection. Screening is carried out in two steps: the first step is to screen out positive cell holes by using ic-ELISA, and the second step is to select the trichlorfon as a standard substance, and to measure the inhibition effect of positive cells by using ic-ELISA. Selecting cell holes with better inhibition on the chlorfenapyr standard, subcloning by adopting a limiting dilution method, and detecting by the same method. The cell line JHJ was obtained by repeating the above steps three times.
(6) Preparation and identification of monoclonal antibodies: taking 8-10 week old BALB/c mice, and injecting 1mL of sterile paraffin oil into the abdominal cavity of each mouse; intraperitoneal injection of 1X 10 per mouse after 7 days 6 Hybridoma cells, ascites was collected from the seventh day, and the ascites was purified by the octanoic acid-ammonium sulfate method. Under the condition of meta-acid, the n-octanoic acid can precipitate other hetero proteins except IgG immunoglobulin in ascites, and then the mixture is centrifuged and the precipitate is discarded; precipitating IgG type monoclonal antibody with equal amount of saturated ammonium sulfate solution, centrifuging, discarding supernatant, dissolving with 0.01M PBS solution (pH 7.4), dialyzing for desalting, and storing at-20deg.C;
immunoglobulin subtype identification is carried out on the trichloro acaricidal alcohol monoclonal antibody obtained by ascites purification by using a mouse monoclonal antibody subtype identification kit, the subtype is IgG2b type, and the light chain type is KAPPA type detected by the mouse monoclonal antibody subtype identification kit, as shown in figure 1.
EXAMPLE 2 IC of a monoclonal antibody to Trichloroacarid 50 Is (are) determined by
1, coating: the coating stock prepared in step (2) of example 1 was diluted in a 1. Mu.g/mL ratio with 0.05M carbonate buffer at pH9.6, 100. Mu.L/well, and reacted at 37℃for 2 hours;
2, washing: pouring the solution in the plate, and washing with the washing liquid for 3 times each for 3min;
and 3, closing: after drying, 200. Mu.L/well of blocking solution was added thereto and reacted at 37℃for 2 hours. Washing and drying for later use;
4, sample adding: the antisera was diluted from 1:1000 in a double ratio and added to the coated wells at each dilution, 100. Mu.L/well, and reacted at 37℃for 30min; after extensive washing, HRP-goat anti-mouse IgG diluted 1:3000 was added, 100. Mu.L/well, and reacted at 37℃for 30min;
5 developing: taking out the ELISA plate, fully washing, adding 100 mu L of TMB color developing solution into each hole, and carrying out light-shielding reaction for 15min at 37 ℃;
6 termination and measurement: 50. Mu.L of stop solution was added to each well to terminate the reaction, and the OD 450 value of each well was measured by using a microplate reader.
Sensitivity to chlorfenapyr was detected by ic-ELISA as shown in FIG. 2, according to the standard equation y=0.1159+1.375/(1+ (x/1.240) 0.784 ) IC for determining monoclonal antibody of trichlorfon by IC-ELISA 50 The sensitivity of the antibody is 2ng/mL, which indicates that the antibody has higher sensitivity and can be used for the immune analysis and detection of the chlorfenapyr.
The foregoing is only a preferred embodiment of the invention, it being noted that: it will be apparent to those skilled in the art that various modifications and adaptations can be made without departing from the principles of the present invention, and such modifications and adaptations are intended to be comprehended within the scope of the invention.
Claims (5)
1. The hybridoma cell strain secreting the chlorfenapyr monoclonal antibody is preserved in China general microbiological culture Collection center (CGMCC), the institute of microbiological research, national academy of sciences, no. 3, north Chen West Lu No. 1, the area of the Beijing, and the dynasty, and is classified and named as monoclonal cell strain JHJ, the preservation date is 2021, the month is 05, and the preservation number is CGMCC No.22329.
2. The hybridoma cell line of claim 1, wherein said hybridoma cell line secretes and produces a monoclonal antibody to chlorfenapyr.
3. The hybridoma cell strain of claim 2, wherein the trichlorfon monoclonal antibody is used for analysis and detection of trichlorfon residues in food safety detection.
4. The hybridoma cell line according to claim 3, wherein the monoclonal antibody against trichlorfon is used for preparing a test body for analysis and detection of residual trichlorfon in food safety test.
5. The hybridoma cell line according to claim 4, wherein the detection body comprises a reagent, a detection plate, and a kit.
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