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Anti-phenacetin monoclonal antibody hybridoma cell strain AD as well as preparation method and application thereof

A technology for hybridoma cell lines and cloning antibodies, applied in biochemical equipment and methods, carboxylic acid amide preparation, chemical instruments and methods, etc., to achieve good detection sensitivity and specificity

Active Publication Date: 2021-06-01
JIANGNAN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, there is no relevant report on the use of enzyme-linked immunosorbent assay for phenacetin detection so far

Method used

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  • Anti-phenacetin monoclonal antibody hybridoma cell strain AD as well as preparation method and application thereof
  • Anti-phenacetin monoclonal antibody hybridoma cell strain AD as well as preparation method and application thereof
  • Anti-phenacetin monoclonal antibody hybridoma cell strain AD as well as preparation method and application thereof

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Embodiment

[0027] The present invention can be better understood from the following examples. However, those skilled in the art will readily understand that the specific material ratios, process conditions and results described in the examples are only used to illustrate the present invention, and should not and will not limit the present invention described in detail in the claims .

[0028] The present invention immunizes mice with phenacetin complete antigen, through cell fusion, HAT selective medium culture, and icELISA screening of cell supernatant, and finally obtains a monoclonal antibody hybridoma with high sensitivity to phenacetin cell line.

[0029] 1. Hapten Synthesis

[0030] (1) Synthesis of hapten Phe-BA:

[0031] The synthetic route is as follows:

[0032]

[0033] Dissolve 0.20 g of acetaminophen in 2 mL of dimethyl sulfoxide, add 1.0 g of K 2 CO 3 , stirred for 0.5h. Then, 0.77 g of ethyl 4-bromobutyrate was added under stirring, heated to reflux, stirred at 1...

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Abstract

The invention discloses an anti-phenacetin monoclonal antibody hybridoma cell strain AD as well as a preparation method and an application thereof, and relates to the technical field of food safety immunodetection, the monoclonal antibody hybridoma cell strain has a preservation name of monoclonal cell strain AD and a preservation number of CGMCC19681. A reaction product of phenacetin metabolite acetaminophen and ethyl 4-bromobutyrate is hydrolyzed to obtain Phe-BA as a hapten, and the hapten and carrier protein are coupled to prepare the immunogen Phe-BA-BSA. After a mouse is subjected to immunogen Phe-BA-BSA injection immunization, the mouse is fused with myeloma cells through a PEG method, and the hybridoma cell strain is obtained through screening and five times of subcloning of an indirect competitive enzyme-linked immunosorbent assay. The monoclonal antibody secreted by the cell strain can be prepared into a phenacetin detection kit, has good affinity and detection sensitivity to phenacetin, and can be used for immunodetection of phenacetin residual quantity in food.

Description

technical field [0001] The invention belongs to the technical field of food safety immunoassay, and in particular relates to an anti-phenacetin monoclonal antibody hybridoma cell line AD and a preparation method and application thereof. Background technique [0002] Phenacetin (Phe) belongs to nonsteroidal antiinflammatory drugs (Nonsteroidal Antiinflammatory Drugs, NSAIDs), has anti-inflammatory, analgesic, antipyretic and other effects, and is widely used clinically in the treatment of various fever and various pain symptoms. Relief. At present, NSAIDs are one of the most used drugs in the world. There are about 30 million people in the world who use them every day. With the increase in the use of NSAIDs, the safety use of these drugs has also attracted more and more attention. The U.S. Food and Drug Administration The Food and Drug Administration (FDA) believes that NSAIDs have potential cardiovascular and gastrointestinal bleeding risks, which makes the safe use of NSAID...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N5/20C07K16/44C07C231/12C07C233/25G01N33/577G01N33/53C12R1/91
CPCC07K16/44C07C231/12G01N33/9486G01N33/577C07K2317/92C07C233/25G01N2470/10
Inventor 胥传来姚静静匡华刘丽强宋姗姗吴爱红徐丽广胡拥明
Owner JIANGNAN UNIV
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