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Chicken interleukin-18 monoclonal antibody as well as preparation method and application thereof

A monoclonal antibody and interleukin technology, which is applied to the preparation method of peptide, interleukin, biochemical equipment and methods, etc., can solve the problems of low titer, poor specificity of monoclonal antibody, and restrictions on production and application

Inactive Publication Date: 2014-02-19
HARBIN VETERINARY RES INST CHINESE ACADEMY OF AGRI SCI +1
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0003] At present, the hybridoma cell lines used to secrete chicken interleukin-18 monoclonal antibody have defects such as low titer and poor stability, and the specificity of the secreted monoclonal antibody is poor, which seriously restricts production and application, and needs to be improved urgently

Method used

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  • Chicken interleukin-18 monoclonal antibody as well as preparation method and application thereof
  • Chicken interleukin-18 monoclonal antibody as well as preparation method and application thereof
  • Chicken interleukin-18 monoclonal antibody as well as preparation method and application thereof

Examples

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Embodiment 1

[0023] Example 1 Preparation and Identification of Chicken Interleukin-18 Monoclonal Antibody

[0024] 1. Experimental method

[0025] 1.1 Immunization of mice

[0026] (1) Prepare SDS-PAGE strips containing ChIL-18 recombinant protein;

[0027] (2) Before immunization, add a small amount of PBS and grind it with a grinding rod, and add an appropriate amount of double antibody solution, preferably the colloidal particles can pass through the needle of a 1mL syringe, that is, the immunogen;

[0028] (3) Select 5 clean-grade BALB / c mice aged 6-8 weeks, and immunize them with the prepared immunogen by intraperitoneal injection, 300 μL each time, and immunize once every 2 weeks;

[0029] (4) One week after the third immunization, blood was collected from the infraorbital sinus, and then serum was separated according to conventional methods;

[0030] (5) Perform IFA verification test on the antiserum of immunized mice and CHO-ChIL-18F cells to detect whether the two can react, a...

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Abstract

The invention discloses a chicken interleukin-18 monoclonal antibody as well as a preparation method and an application thereof. According to the method, recombinant protein ChIL-18 for prokaryotic expression is taken as an immunogen, and ChIL-18 protein for eukaryotic cell line expression is taken as a screening source; and after cell fusion, hybridoma cells are screened with an IFA (indirect immunofluorescence assay), and eight cell strains which secrete the chicken interleukin-18 monoclonal antibody are obtained through screening, wherein the stability of a hybridoma cell strain 4E11 is the best, the titer and the specificity of the monoclonal antibody secreted by the hybridoma cell strain 4E11 are the highest, and the microbial preservation number is CGMCC NO. 7658. The invention further provides the hybridoma cell strain 4E11 and an application of the monoclonal antibody secreted by the hybridoma cell strain 4E11 in detection or purification of the chicken interleukin-18 protein.

Description

technical field [0001] The present invention relates to a chicken interleukin monoclonal antibody, in particular to a chicken interleukin-18 monoclonal antibody and a hybridoma cell line secreting the monoclonal antibody. The present invention further relates to the application of the monoclonal antibody in detecting or purifying chicken interleukin-18 protein The invention belongs to the field of preparation and application of chicken interleukin-18 monoclonal antibody. Background technique [0002] Interleukin, referred to as interleukin, is a cytokine produced by white blood cells that mediates intercellular interactions and plays a regulatory role in cell activation, proliferation and differentiation. With the development of molecular biology, research on chicken interleukin genes and functions has gradually become a hot spot. So far, a variety of interleukins have been isolated from chickens, such as: IL-1, IL-2, IL-6, IL- 10. IL-12, IL-16, IL-18, etc. These interleuki...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N5/20C07K16/24G01N33/68G01N33/577C07K14/54C07K1/22C12R1/91
Inventor 韩凌霞文辉强廉传江司昌德林欢陈洪岩
Owner HARBIN VETERINARY RES INST CHINESE ACADEMY OF AGRI SCI
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