CN-122012406-A - Hybridoma cell strain secreting azaphenanthridine monoclonal antibody and application thereof

Abstract

The invention relates to a hybridoma cell strain secreting an azaphenanthridine monoclonal antibody and application thereof, and belongs to the technical field of safety immunodetection. The hybridoma cell strain is preserved in China general microbiological culture Collection center (China Committee for culture Collection of microorganisms) at the date of 11 and 12 of 2025, and has a preservation address of North Star Xiyu No.1, 3 in the Korean region of Beijing city and a preservation number of CGMCC No.46743. The azaphenanthridine monoclonal antibody is secreted by the hybridoma cell strain, has good detection sensitivity (IC 50 value is 0.529 ng/mL) on the azaphenanthridine, and can be used for detecting the residue of the azaphenanthridine.

Inventors

• XU CHUANLAI
• WU AIHONG
• GUO LINGLING
• XU XINXIN
• QU AIHUA
• Kou shuai
• KUANG HUA
• XU LIGUANG
• SUN MAOZHONG
• WU XIAOLING
• LIU LIQIANG
• HAO CHANGLONG
• SONG SHANSHAN

Assignees

• 江南大学

Dates

Publication Date

20260512

Application Date

20260129

Claims (10)

1. 1. The hybridoma cell strain secreting the azaphenanthridine monoclonal antibody is characterized in that the hybridoma cell strain is preserved in China general microbiological culture Collection center (China Committee for culture Collection of microorganisms) on the 11 th month of 2025, the preservation address is North Star Xiyu No.1, 3 in the Korean region of Beijing city, and the preservation number is CGMCC No.46743.
2. 2. An azaphenanthridine monoclonal antibody produced by secretion from the hybridoma cell line of claim 1.
3. 3. The azaphenanthridine monoclonal antibody according to claim 1, wherein the azaphenanthridine monoclonal antibody is obtained from an azaphenanthridine complete antigen immunized animal.
4. 4. The azaphenanthridine monoclonal antibody according to claim 3, wherein the complete antigen of azaphenanthridine is obtained by coupling the azaphenanthridine hapten to a carrier protein.
5. 5. The azaphenanthridine monoclonal antibody according to claim 4, wherein the azaphenanthridine hapten has the following structural formula: 。
6. 6. The azaphenanthridine monoclonal antibody according to claim 4, wherein the carrier protein comprises bovine serum albumin and/or chicken ovalbumin.
7. 7. A composition comprising the hybridoma cell line of claim 1 and/or the azaphenanthridine monoclonal antibody of any one of claims 2-6.
8. 8. A kit comprising one or more of the hybridoma cell line of claim 1, the azaphenanthridine monoclonal antibody of any one of claims 2-6, and the composition of claim 7.
9. 9. A test strip comprising one or more of the hybridoma cell line of claim 1, the azaphenanthridine monoclonal antibody of any one of claims 2-6, and the composition of claim 7.
10. 10. Use of the hybridoma cell line of claim 1, the azaphenanthridine monoclonal antibody of any one of claims 2-6, the composition of claim 7, the kit of claim 8 or the test strip of claim 9 for detecting azaphenanthridine, wherein the use does not involve diagnosis and treatment of diseases.

Description

Hybridoma cell strain secreting azaphenanthridine monoclonal antibody and application thereof Technical Field The invention relates to the technical field of safety immunodetection, in particular to a hybridoma cell strain secreting an azaphenanthridine monoclonal antibody and application thereof. Background Azophenanthridine (isometamidium, ISM) is a long-acting anti-trypanosome drug composed of a group of compound isomers, and is widely applied to prevention and treatment of animal parasitic diseases such as cattle and sheep as a chemical drug for preventing and treating animal sleeping diseases by inhibiting trypanosome DNA and RNA polymerase and preventing nucleic acid synthesis. However, the azaphenanthridine has side effects of cancerogenesis, malformation, mutation and the like while preventing and treating parasitic diseases of animals of cattle and sheep. The abuse of the prevention and treatment medicines can not only affect animals per se, but also cause potential harm to human health due to residues in animal foods. To mitigate the negative effects of azaphenanthridine on human health, many countries have established a Maximum Residual Limit (MRL) in animal-derived products. In China, the highest residual limit of azaphenanthridine is set to 0.1 mg/kg. Therefore, detecting the residual amount of the azaphenanthridine in the animal-derived agricultural products is important for guaranteeing the health of human bodies. Currently, the detection method of the azaphenanthridine is mainly instrument detection, and commonly used methods include 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). Although these chromatographic methods have high sensitivity and specificity, there are also disadvantages such as thorough purification of the sample, high solvent consumption, expensive equipment, high requirements on the expertise of the operator, and the like. Therefore, it is highly desirable to establish a rapid, simple method for detecting the residual amount of azaphenanthridine. The ELISA is a high-efficiency, sensitive and rapid detection method, and has the advantages of simple sample pretreatment requirement, less purification steps, high analysis flux, low detection cost and simple and convenient operation, and is suitable for the on-site rapid detection of a large number of samples, thereby being widely applied to the field of drug residue analysis. On the premise of detecting the azaphenanthridine by adopting an enzyme-linked immunosorbent assay, the monoclonal antibody with high specificity and high sensitivity to the azaphenanthridine is obtained. Therefore, it is important to establish a preparation method of the high-specificity and high-sensitivity monoclonal antibody of the azaphenanthridine. The inventor tries to prepare the azaphenanthridine monoclonal antibody by a hybridoma cell technology, but in the process of preparing a hybridoma cell strain capable of secreting the azaphenanthridine monoclonal antibody, a plurality of key problems still need to be intensively studied, namely, a preparation method of an azaphenanthridine hapten and a complete antigen, how to enable a mouse to generate a strong immune response effect, how to ensure that the prepared hybridoma cell strain can stably secrete the azaphenanthridine monoclonal antibody, and how to improve the specificity and sensitivity of the secreted monoclonal antibody. Disclosure of Invention In order to solve the technical problems, the invention provides a hybridoma cell strain secreting an azaphenanthridine monoclonal antibody and application thereof. The azaphenanthridine monoclonal antibody secreted by the hybridoma cell strain has good detection sensitivity (IC 50 values are 0.529 ng/mL respectively) to the azaphenanthridine, and can be used for establishing an immunological detection method of the azaphenanthridine and detecting the residue of the azaphenanthridine in food. The invention is realized by the following technical scheme: The first object of the present invention is to provide a hybridoma cell strain secreting an azaphenanthridine monoclonal antibody, wherein the hybridoma cell strain is preserved in the China general microbiological culture Collection center (China Committee for culture Collection of microorganisms) at the year 2025, 11 and 12, and the preservation address is the Seagaku No. 1, 3 in the Korean region North Star in Beijing city, and the preservation number is CGMCC No.46743. The second object of the invention is to provide an azaphenanthridine monoclonal antibody which is secreted by the hybridoma cell strain. In one embodiment of the invention, the azaphenanthridine monoclonal antibody is obtained by immunizing an animal with an azaphenanthridine complete antigen. Specifically, BALB/c mice are