CN-121975741-A - Hybridoma cell strain secreting rofecoxib monoclonal antibody and application thereof

Abstract

The invention relates to a hybridoma cell strain secreting a rofecoxib monoclonal antibody and application thereof, and belongs to the technical field of immunodetection. The invention prepares the complete antigen of rofecoxib, and screens to obtain a hybridoma cell strain secreting the monoclonal antibody of the rofecoxib through the steps of animal immune reaction and cell fusion, and the hybridoma cell strain is preserved in China general microbiological culture Collection center (CGMCC) with the preservation number of 46734. The monoclonal antibody secreted by the hybridoma cell strain has good sensitivity and specificity, has an IC 50 value of 20.0 ng/mL for rofecoxib, has cross reaction rates of less than 1% for various rofecoxib functional analogues, can be applied to preparing a rofecoxib detection product, and provides an efficient detection method and means for detecting the residual of the rofecoxib in food.

Inventors

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

Assignees

• 江南大学

Dates

Publication Date

20260505

Application Date

20251212

Claims (10)

1. 1. The hybridoma cell strain secreting the rofecoxib monoclonal antibody is characterized in that the hybridoma cell strain is preserved in China general microbiological culture Collection center (China general microbiological culture Collection center) at the year 2025, the preservation address is number 3 of the West-Lou 1, the Korean region North Star in Beijing, and the preservation number is CGMCC No.46734.
2. 2. The hybridoma cell line according to claim 1, wherein the hybridoma cell line is obtained from a rofecoxib hapten by synthesizing a rofecoxib complete antigen and immunizing an animal.
3. 3. The hybridoma cell line of claim 1, wherein the rofecoxib hapten has the structural formula: 。
4. 4. A monoclonal antibody of rofecoxib is characterized in that, the rofecoxib monoclonal antibody is secreted by the hybridoma cell line of any one of claims 1-3.
5. 5. A composition comprising the hybridoma cell line of any one of claims 1-3 and/or the rofecoxib monoclonal antibody of claim 4.
6. 6. A test strip for detecting rofecoxib, wherein the test strip comprises the hybridoma cell strain of any one of claims 1-3, the rofecoxib monoclonal antibody of claim 4, or the composition of claim 5.
7. 7. The test strip of claim 6, further comprising a sample pad, a binding pad, a water absorbing pad, and a cellulose acetate membrane, wherein the binding pad is coated with a rofecoxib monoclonal antibody, and the rofecoxib monoclonal antibody is labeled with colloidal gold or fluorescent microspheres.
8. 8. A kit for detecting rofecoxib, comprising the hybridoma cell line of any one of claims 1-3, the rofecoxib monoclonal antibody of claim 4, or the composition of claim 5.
9. 9. The kit according to claim 8, further comprising an ELISA plate, a rofecoxib coating antigen, a rofecoxib standard solution, an enzyme-labeled secondary antibody and a color development solution.
10. 10. Use of the hybridoma cell line of any one of claims 1-3, the rofecoxib monoclonal antibody of claim 4, the composition of claim 5, the test strip of claim 6 or 7 or the kit of claim 8 or 9 for detecting rofecoxib, wherein the use does not involve diagnosis or treatment of a disease.

Description

Hybridoma cell strain secreting rofecoxib 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 rofecoxib monoclonal antibody and application thereof. Background Rheumatic and rheumatoid osteoarthritis are common diseases and frequently encountered diseases of middle-aged and elderly people, and are chronic diseases which can not be radically treated, and the symptoms of joint swelling and pain seriously affect the life quality of patients. Clinically, anti-inflammatory analgesic drugs are the first therapeutic scheme for alleviating such symptoms, and the action mechanism is mainly to reduce the generation of prostaglandin by inhibiting cyclooxygenase, thereby achieving the purpose of diminishing inflammation and relieving pain. Non-steroidal anti-inflammatory drugs (NSAIDs) are the mainstream of such drugs, and can be classified into acid derivatives (e.g., aspirin), oxicams (e.g., meloxicam), sibs (e.g., rofecoxib, celecoxib), pyrazoles (e.g., phenylbutazone), and the like according to structures. However, NSAIDs may also cause a series of serious toxic side effects, including gastrointestinal damage, blood system damage, hepatorenal toxicity, elevated blood pressure, anaphylaxis, and nervous system side effects, while exerting therapeutic effects. As such, its use must be under strict medical supervision. However, in recent years, the phenomenon of illegally adding such chemical drugs to traditional Chinese medicines and health foods has occurred. In order to make the products show the false appearance of quick-acting and high-efficiency, some illegal manufacturers cheat consumers and gain violence, and anti-inflammatory analgesics such as rofecoxib and the like are illegally added, so that huge medication safety risks are brought to consumers with unknown and real states. To combat this disorder, the regulatory authorities have established corresponding detection methods, such as the 2009 national food and drug administration approved supplementary test methods for antirheumatic Chinese patent drugs. However, the method and the TLC, HPLC, HPLC-MS/MS reported in the literature have the limitations of complicated operation, time and labor waste, expensive instruments and the like, and are difficult to meet the urgent requirement of rapid and on-site screening of a large number of samples in market supervision. Huang Yueyan and the like, determining 17 anti-inflammatory analgesic chemical drugs including rofecoxib in traditional Chinese medicines and health-care foods by utilizing an ultra-high performance liquid chromatography-tandem mass spectrometry, determining the added chemical drugs by comparing information such as molecular ion peaks, secondary fragment ion peaks, chromatographic retention time and the like of a sample peak and a reference peak in an MRM channel, and calculating the accurate quantity of the added drugs according to an external standard method by using mass spectrum peak areas. (UPLC-MS/MS method for rapidly determining 17 anti-inflammatory analgesic chemical drugs illegally added in Chinese medicinal materials and health food [ J ]. Chinese herbal medicine 2016,47 (02): 246-254.). The detection Limit (LOD) of the 17 anti-inflammatory analgesic chemical methods is 0.3-5.0 ng/g, the quantitative Limit (LOQ) is 0.9-15.0 ng/g, and the sample recovery rate is 90.5% -113.8%. The method is complex in operation, complex in sample treatment, unfavorable for detection of a large number of samples in actual production, urgent in need of establishing a detection method with simple operation, high sensitivity and strong specificity, is used for effectively monitoring illegal addition of rofecoxib in health-care food, and has important significance for guaranteeing public health and market order. Disclosure of Invention In order to solve the technical problems, the invention provides a hybridoma cell strain secreting a rofecoxib monoclonal antibody and application thereof. The invention prepares the complete antigen of rofecoxib by synthesizing the hapten of rofecoxib, screens to obtain a hybridoma cell strain secreting the monoclonal antibody of rofecoxib through the steps of mouse immune reaction and cell fusion, and is preserved in the China general microbiological culture Collection center (CGMCC) No.46734. The invention is realized by the following technical scheme: The first object of the present invention is to provide a hybridoma cell strain secreting rofecoxib monoclonal antibody, wherein the hybridoma cell strain is preserved in China general microbiological culture Collection center (China general microbiological culture Collection center) at the 11 th month and 12 th year of 2025, and the preservation address is the North Star Xiya No. 1, 3 in the Korean region of Beijing city, and the preservation number is CGMCC No.46734. In one embodiment of the invention, t