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Hybridoma cell strain ZXL-1 secreting specific monoclonal antibody to HB (hygromycin B) and application of hybridoma cell strain ZXL-1

A hybridoma cell line and monoclonal antibody technology, applied in the field of immunoassay, can solve the problems of cumbersome and time-consuming processing

Active Publication Date: 2017-09-19
JIANGNAN UNIV +1
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  • Abstract
  • Description
  • Claims
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Problems solved by technology

[0004] At present, the most commonly used detection methods for HB are instrumental detection methods such as high performance liquid chromatography. This detection method takes a long time, requires relevant professional skills for operation, and cumbersome sample pretreatment, etc.

Method used

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  • Hybridoma cell strain ZXL-1 secreting specific monoclonal antibody to HB (hygromycin B) and application of hybridoma cell strain ZXL-1

Examples

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Effect test

Embodiment 1

[0037] Example 1 Preparation of HB Monoclonal Antibody

[0038]1. Preparation of immunogen and coating source: the immunogen HB-BSA obtained by coupling HB with bovine serum albumin (BSA) by the glutaraldehyde method; The coupled HB-OVA was used as the coating antigen.

[0039] The specific synthesis steps of HB-BSA are as follows:

[0040] a. Weigh 10 mg of HB, dissolve it in 1 ml of 0.1 M phosphate buffer solution with pH 7.4, then add 100 μL of glutaraldehyde solution with a mass fraction of 25% diluted 10 times dropwise, and stir at room temperature for 30 minutes to obtain an activation solution ;

[0041] b. Weigh 50mg of BSA, dissolve it in 5mL 0.1M pH9.0 carbonate buffer solution, add the activation solution drop by drop under constant stirring, and react for 3-4 hours at room temperature, then dilute the reaction solution with 0.01M pH 7.4 Phosphate buffered saline dialyzed to obtain HB-BSA.

[0042] The specific synthesis steps of HB-OVA are as follows:

[0043]...

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Abstract

The invention discloses a hybridoma cell strain ZXL-1 secreting a specific monoclonal antibody to HB (hygromycin B) and an application of the hybridoma cell strain ZXL-1 and belongs to the technical field of immunodetection. The hybridoma cell strain ZXL-1 secreting the specific monoclonal antibody to the HB has been collected in CGMCC (China general microbiogical culture collection center) of CCCCM (China committee for culture collection of microorganisms) with the collection number of CGMCC No. 13097. The anti-HB monoclonal antibody is obtained from the obtained hybridoma cell strain ZXL-1 specifically secreting the antibody to the HB after ascites preparation and purification. The antibody can be used for immunodetection of HB residues in animal-derived food and meet the market requirement for quick HB immunodetection products at present.

Description

technical field [0001] The invention relates to a hybridoma cell line ZXL-1 secreting hygromycin B specific monoclonal antibody and its application, to a preparation method of hygromycin B monoclonal antibody, and to the indirect competition of hygromycin B in animal source food The establishment of an enzyme-linked immunoassay method belongs to the technical field of immunoassays. Background technique [0002] Hygromycin B (HB) is an aminoglycoside antibiotic produced by Streptomyces hygroscopicus, which can kill bacteria, fungi and advanced eukaryotic cells by inhibiting protein synthesis. It has a certain deworming activity, and it can be added to pig and poultry feed for a long time, and has a good effect of repelling nematodes. [0003] The problem of veterinary drug residues in food of animal origin is one of the public health hotspots in the international community in recent years, and has attracted widespread attention. Since HB has been clinically proven to be oto...

Claims

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Application Information

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IPC IPC(8): C12N5/20C07K16/44G01N33/577G01N33/543
Inventor 胥传来郭玲玲匡华徐丽广马伟刘丽强宋珊珊吴晓玲郑乾坤
Owner JIANGNAN UNIV
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