CN-122012405-A - Hybridoma cell strain secreting bisbenzoxazole monoclonal antibody and application thereof

Abstract

The invention relates to a hybridoma cell strain secreting a bisbenzoxazole monoclonal antibody and application thereof, and belongs to the technical field of immunodetection. The hybridoma cell strain capable of secreting the bisbenzoxazole acid monoclonal antibody is screened for the first time, the monoclonal antibody separated from the hybridoma cell strain has good sensitivity and specificity to the bisbenzoxazole acid, the IC 50 value is 0.2579 ng/mL, and the monoclonal antibody has no cross reaction to structural analogues of the bisbenzoxazole acid such as DDT, fenpyrad, benazolin, fenbuconazole, pyribenzoxim and the like. Therefore, the monoclonal antibody of the invention can be used for establishing an immunological detection method of the bisbenzoxazole acid so as to detect the residue of the bisbenzoxazole acid in food.

Inventors

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

Assignees

• 江南大学

Dates

Publication Date

20260512

Application Date

20260120

Claims (10)

1. 1. The hybridoma cell strain is named as a monoclonal cell strain, and the preservation number is CGMCC No.46735.
2. 2. The hybridoma cell line according to claim 1, wherein the hybridoma cell line is obtained by immunizing an animal with a complete antigen prepared from a hapten, wherein the hapten has the structural formula shown in formula I: 。
3. 3. the hybridoma cell line according to claim 2, wherein said complete antigen is derived from said hapten-coupled carrier protein.
4. 4. The hybridoma cell line of claim 3, wherein said carrier protein comprises keyhole limpet hemocyanin.
5. 5. Use of the hybridoma cell line of any one of claims 1-4 for detecting bisbenzoxazole acid.
6. 6. A monoclonal antibody secreted by the hybridoma cell line of any one of claims 1-4.
7. 7. The use of the monoclonal antibody of claim 6 for detecting bisbenzoxazole acid.
8. 8. A detection product of bisbenzoxazole acid, characterized in that said detection product comprises the monoclonal antibody of claim 6.
9. 9. The test product of claim 8, further comprising a coating antigen.
10. 10. The test product of claim 9, wherein the coating antigen is made from hapten-coupled carrier proteins, wherein the carrier proteins comprise chicken ovalbumin.

Description

Hybridoma cell strain secreting bisbenzoxazole monoclonal antibody and application thereof Technical Field The invention relates to the technical field of immunodetection, in particular to a hybridoma cell strain secreting a bisbenzoxazole monoclonal antibody and application thereof. Background The bisbenzoxazole acid is a herbicide safener which is specially used for protecting crops from herbicide injury, is mainly used in corn and paddy fields, and can protect crops and improve the weeding effect. The research shows that it can up-regulate the activity of cytochrome P450 monooxygenase, glutathione-S-transferase, hydrolase and other key enzyme system in crop, so as to speed up the oxidation, conjugation and hydrolysis reaction of herbicide molecule, convert it into non-toxic or low-toxic product and isolate the product for storage. It can enter the human body through skin contact, inhalation or misfeeding. Environmental residue and toxicity, namely that the bisbenzoxazole acid is easy to remain in soil and water, has extremely high toxicity to aquatic organisms (such as fish) and also forms threat to non-target organisms such as bees, birds and the like. Its strong mobility may also pose a pollution risk to groundwater. Since bisbenzoxazole acid has a long durability, it can be accumulated in soil. Furthermore, it may cause long term adverse effects in aquatic environments. While these materials are classified by the U.S. environmental protection agency as preferential toxic pollutants, they are used in their pure form or in combination with other materials in many parts of the world for various herbicides. In order to mitigate the negative effects of bisbenzoxazole acid on human health, many countries have established the Maximum Residual Limit (MRL) for its use in agricultural products. EU 396/2005 regulation in the European Union sets the highest residual limit of 0.05 mg/kg of bisbenzoxazole acid. Detecting the presence of the residue of bisbenzoxazole acid in agricultural products is critical to ensuring human health. At present, the method for detecting the bisbenzoxazole acid 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 combined tandem mass spectrometry (UPLC-MS/MS). Although these chromatographic-based methods have high sensitivity and specificity, there are drawbacks such as the need for thorough sample purification, high solvent consumption, expensive equipment and skilled technicians. Thus, there is a need for a rapid, simple method of detecting bisbenzoxazole acid residues. The ELISA is a very efficient, sensitive and rapid detection method, and has the advantages of simple pretreatment of samples during detection, few purification steps, large analysis capacity, low detection cost and simple and convenient operation, and is suitable for the on-site rapid detection of a large number of samples, thus being widely applied to the analysis of drug residues. On the premise that the enzyme-linked immunosorbent assay is used for detecting the bisbenzoxazole acid, a monoclonal antibody with high specificity and high sensitivity to the bisbenzoxazole acid is obtained. Such a monoclonal antibody is lacking in the prior art. Disclosure of Invention Therefore, the technical problem to be solved by the invention is to overcome the problem that a high-specificity and high-sensitivity bisbenzoxazole monoclonal antibody is lacking in the prior art. In order to solve the technical problems, the invention provides a hybridoma cell strain secreting a bisbenzoxazole monoclonal antibody and application thereof. The hybridoma cell strain capable of secreting the bisbenzoxazole acid monoclonal antibody is screened for the first time, the monoclonal antibody separated from the hybridoma cell strain has good sensitivity and specificity to the bisbenzoxazole acid, the IC 50 value is 0.2579 ng/mL, and the monoclonal antibody has no cross reaction to structural analogues of the bisbenzoxazole acid such as DDT, fenpyrad, benazolin, fenbuconazole, pyribenzoxim and the like. Therefore, the monoclonal antibody of the invention can be used for establishing an immunological detection method of the bisbenzoxazole acid so as to detect the residue of the bisbenzoxazole acid in food. The first object of the present invention is to provide a hybridoma cell strain, which is characterized in that the hybridoma cell strain is named as a monoclonal cell strain, and the preservation number is CGMCC No.46735. Further, the hybridoma cell strain is obtained by immunizing an animal with a complete antigen prepared from a hapten, wherein the structural formula of the hapten is shown as formula I: 。 further, the complete antigen is derived from the hapten-coupled carrier protein. Further, the carrier pro