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Chloramphenicol universal monoclonal antibody hybridoma cell strain and application thereof

A monoclonal antibody, chloramphenicol technology, applied in biochemical equipment and methods, instruments, microorganisms, etc., can solve the problems of expensive instruments, complicated operations, and long time.

Active Publication Date: 2015-01-07
JIANGNAN UNIV
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

Among them, the instrument detection method is accurate, but the instrument is expensive,

Method used

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  • Chloramphenicol universal monoclonal antibody hybridoma cell strain and application thereof

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Embodiment 1

[0034] Example 1 Preparation of chloramphenicol-like universal monoclonal antibodies

[0035] 1. Preparation of complete antigen

[0036] Preparation of solution A: take 0.023mmol hapten TAP-NH 2 In a 25mL beaker, add 2mL DMF, 0.2mL water and 0.2mL 1mol / L hydrochloric acid solution to form a yellow solution, and cool to 0-5°C. Then add 1mol / L sodium nitrite solution drop by drop, monitor with starch-potassium iodide test paper, stop dripping until the test paper turns blue-gray, and stir at 4°C for 1h.

[0037] Preparation of liquid B: Weigh 20 mg of KLH, 100 mg of BSA, and 150 mg of OVA to dissolve each in 6 mL of pH 9.0 borate buffer to obtain three different B solutions and store them at 4°C. Under stirring at 4℃, add liquid A to liquid B dropwise, and adjust the pH with 1mol / L sodium hydroxide solution to keep the pH at 9, stir overnight at 4℃, and dialyze to obtain immunogen TAP-NH. 2 -KLH, coated original TAP-NH 2 -BSA and TAP-NH 2 -OVA.

[0038] 2. Animal immunization: select ...

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Abstract

The invention discloses a chloramphenicol universal monoclonal antibody hybridoma cell strain and application thereof, belonging to the technical field of food safety immunological detection. The preparation method comprises the following steps: by taking a thiamphenicol analogue 1-(4-aminophenyl)-2-dichlorethamin-1,3-propylene glycol as a hapten, allowing the hapten to pass through diazotization coupling bovine serum albumin (BSA), thereby obtaining a complete antigen; uniformly mixing with a Freund's adjuvant, and performing subcutaneous injection immunotherapy on BALB/c mice; synthesizing an envelope antigen to be used for screening mice serum and cell supernatant by using diazotization; and fusing splenocyte of immune mice with mouse myeloma cells through a PEG method, performing indirect ELISA and indirect competitive ELISA screening and subcloning for three times, thereby obtaining a hybridoma cell strain with mass selection property. The cell strain provided by the invention has high inhibition properties on thiamphenicol, chloramphenicol and florfenicol, and the requirements on chloramphenicol stumpy immunodetection products in the current market can be met.

Description

Technical field [0001] The invention relates to a chloramphenicol-like universal monoclonal antibody hybridoma cell strain and its application, and specifically relates to a preparation method of chloramphenicol-like universal monoclonal antibody, which belongs to the technical field of food safety immunological detection. Background technique [0002] Chloramphenicol antibiotics are one of the widely used veterinary drugs. Currently, there are mainly three drugs: chloramphenicol (CAP), thiamphenicol (TAP) and florfenicol (FF). However, chloramphenicol can cause aplastic anemia in humans. This toxic effect has nothing to do with the dosage and the course of treatment, which has attracted the attention of many countries and regions. The United States only allows CAP to be used in non-edible animals and cannot be detected in animal food; the European Union does not allow CAP to be used in dairy cows and laying hens, and its use in other large animals is also restricted. The Minist...

Claims

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Application Information

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IPC IPC(8): C12N5/20C07K16/44G01N33/577C12R1/91
Inventor 匡华郭玲玲胥传来徐丽广马伟刘丽强宋珊珊吴晓玲
Owner JIANGNAN UNIV
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