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Monoclonal antibody hybridoma cell strain of soybean lipoxygenase Lox3 as well as antibody and application of monoclonal antibody

A hybridoma cell line and monoclonal antibody technology, applied in biochemical equipment and methods, anti-enzyme immunoglobulin, instruments, etc., can solve the problems of sensitivity and specificity limitations

Active Publication Date: 2014-04-02
INST OF CEREAL & OIL CROPS HEBEI ACAD OF AGRI & FORESTRY SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, since it is determined by the principle of color reaction of lipoxygenase coupled redox indicator, its sensitivity and specificity are limited to a certain extent.

Method used

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  • Monoclonal antibody hybridoma cell strain of soybean lipoxygenase Lox3 as well as antibody and application of monoclonal antibody
  • Monoclonal antibody hybridoma cell strain of soybean lipoxygenase Lox3 as well as antibody and application of monoclonal antibody
  • Monoclonal antibody hybridoma cell strain of soybean lipoxygenase Lox3 as well as antibody and application of monoclonal antibody

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0074] Embodiment 1: the establishment of the monoclonal antibody hybridoma cell line of anti-soybean lipoxygenase isozyme

[0075] 1.1 Experimental materials:

[0076] Soybean lipoxygenase complete deletion material "Zhongte No. 1" was purchased from the Institute of Crop Science, Chinese Academy of Agricultural Sciences;

[0077] The preservation numbers of the National Resource Bank of Century near-isogenic materials in the United States are: Lox-1: WDD01483, Lox-3: WDD01485;

[0078] Lox-2 enzyme-deficient soybean variety "Wuxing No. 2", approval number: National Examination Bean 2004005;

[0079] BALB / C mice and mouse SP2 / 0 cells were purchased from the Experimental Animal Center of Academy of Military Medical Sciences (Beijing);

[0080] New Zealand white rabbits: purchased from the Experimental Animal Center of Hebei Province;

[0081] 96-well polystyrene microplates were purchased from Xiamen Yunpeng Technology Development Co., Ltd.

[0082] 1.2 Chemical reagents: ...

Embodiment 2

[0107] Example 2: Preparation, detection and application of monoclonal antibodies

[0108] 2.1 Detection antibody preparation

[0109] 2.1.1 Monoclonal antibody preparation and purification

[0110] Preparation of monoclonal antibody ascites: the cell line Lox1-E obtained in step ⑧ in Example 1 1 , Lox2-G 5 , Lox3-H 8 Use culture flasks to expand culture respectively, with 1×10 6 The amount of cells / mouse was inoculated into the peritoneal cavity of BALB / C mice pretreated with paraffin oil to induce ascites. Each cell line was inoculated into 5 mice, and the ascites could be collected after 7 days. Lox1-E 1 , Lox2-G 5 , Lox3-H 8 25ml, 19ml, and 23ml of monoclonal antibody ascites were obtained respectively.

[0111] Ascites titer determination of monoclonal antibody: use indirect ELISA method, Sigma company product Soybean Lox type1-B as detection antigen, determine Lox1-E 1 , Lox2-G 5 , Lox3-H 8 The titers of monoclonal antibody ascites were 1:320,000, 1:640,000, an...

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Abstract

The invention discloses a monoclonal antibody hybridoma cell strain of soybean lipoxygenase Lox3, wherein a collection number of the monoclonal antibody hybridoma cell strain is CGMCC5123. The invention further discloses a monoclonal antibody secreted by the hybridoma cell strain and application of the hybridoma cell strain to detection of the soybean lipoxygenase Lox3.

Description

[0001] Case Description [0002] This application is a divisional application of a monoclonal antibody hybridoma cell line and its antibody and application patent application with the application date of August 2, 2011, the application number 201110219334.4, and the invention name is soybean lipoxygenase isozyme. technical field [0003] The present invention relates to a kind of monoclonal antibody hybridoma cell line and its monoclonal antibody, particularly relate to the monoclonal antibody hybridoma cell line and its monoclonal antibody of respectively secreting anti-soybean lipoxygenase isozyme Lox1, Lox2 and Lox3, cell The preservation numbers of the strains are: CGMCC5121, CGMCC5122 and CGMCC5123, which also involve the application of monoclonal antibodies secreted by each of them in the detection of soybean lipoxygenase isozymes by double-antibody sandwich enzyme-linked immunosorbent assay (DAS-ELISA), which belongs to biological detection technology field. Backgroun...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N5/20C07K16/40G01N33/577G01N33/573
Inventor 马志民张孟臣秦君闫龙李亚璞蒋春志张梅申邸锐唐晓东许宁
Owner INST OF CEREAL & OIL CROPS HEBEI ACAD OF AGRI & FORESTRY SCI
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