Hybridoma cell strain that secrets anti-dinitolmide monoclonal antibodies and the application of hybridoma cell strain

Patent metadata

Field	Value
Publication number	US-2022010029-A1
Application number	US-202117368108-A
Country	US
Kind code	A1
Filing date	Jul 6, 2021
Priority date	Jul 8, 2020
Publication date	Jan 13, 2022
Grant date	—

Abstract

A hybridoma cell strain secreting anti-dinitolmide monoclonal antibodies deposited at China General Microbiological Culture Collection Center (CGMCC) at No. 1 West Beichen Road, Chaoyang District, Beijing 100101, China on Nov. 28, 2019, with deposit number of CGMCC No. 19165. It is classified as a monoclonal cell strain. The monoclonal antibody secreted by the hybridoma cell strain DAS3H10 has a good affinity and high sensitivity to dinitolmide. Because of IC 50 to dinitolmide up to 9.01 ng/mL, the monoclonal antibody could be used to prepare dinitolmide immunoassay kit and colloidal gold test strip and provides a powerful detection method and means for the detection of dinitolmide in animal-derived foods.

First claim

What is claimed is:

1 . A hybridoma cell strain secreting anti-dinitolmide monoclonal antibodies and deposited at China General Microbiological Culture Collection Center (CGMCC) at No. 1 West Beichen Road, Chaoyang District, Beijing 100101, China on Nov. 28, 2019, with deposit number of CGMCC No. 19165.

2 . Anti-dinitolmide monoclonal antibody secreted by the hybridoma cell strain of claim 1 .

3 . A method of making complete immunogen of dinitolmide comprising the following steps: dissolving 4.5 mg of 3,5-dinitro-2-methylbenzoic acid, as well as 5.0 mg EDC (1-ethyl-3-(3-dimethylaminopropyl)carbodiimide) and 3.7 mg NHS (N-hydroxyamide) in DMF (N,N-dimethylformamide), stirring and activating at room temperature for 6 hour to obtain an activated solution; dissolving another 15 mg BSA(bovine serum protein) in 3 mL, 0.05M, pH 9.6 carbonate buffer solution; adding the activated solution dropwise to the BSA solution, stirring at room temperature overnight, and then taking out immunogen and dialyzing with PBS for 3 days; and storing the immunogen by aliquoted storage at −20° C. and obtaining the complete immunogen of dinitolmide.

4 . A method of detecting dinitolmide residues in food comprising applying the anti-dinitrotoamine monoclonal antibody of claim 2 to food and detecting dinitolmide residues in food based on the interaction of the anti-dinitrotoamine monoclonal antibody with the food.

Assignees

• Univ Jiangnan

Inventors

• Xu Chuanlai
• Liu Jie
• Kuang Hua
• Xu Liguang
• Sun Maozhong
• Liu Liqiang
• Wu Xiaoling
• Hao Changlong
• Song Shanshan
• Hu Yongming
• Zheng Qiankun

Classifications

• G01N33/5308

  for analytes not provided for elsewhere, e.g. nucleic acids, uric acid, worms, mites · CPC title

• G01N33/94

  involving narcotics {or drugs or pharmaceuticals, neurotransmitters or associated receptors} · CPC title

• C07K2317/92

  Affinity (KD), association rate (Ka), dissociation rate (Kd) or EC50 value · CPC title

• C07K16/44 Primary

  against material not provided for elsewhere {, e.g. haptens, metals, DNA, RNA, amino acids} · CPC title

• C07K14/795

  Porphyrin- or corrin-ring-containing peptides · CPC title