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Monoclonal antibody hybridoma cell lines of soybean lipoxygenase isozyme, and antibody and application thereof
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A hybridoma cell line, monoclonal antibody technology, applied in biochemical equipment and methods, anti-enzyme immunoglobulin, instruments, etc., can solve problems such as sensitivity and specificity limitations
Active Publication Date: 2013-03-13
INST OF CEREAL & OIL CROPS HEBEI ACAD OF AGRI & FORESTRY SCI
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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.
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Embodiment 1
[0072] Embodiment 1: the establishment of the monoclonal antibody hybridoma cell line of anti-soybean lipoxygenase isozyme
[0073] 1.1 Experimental materials:
[0074] Soybean lipoxygenase complete deletion material "Zhongte No. 1" was purchased from the Institute of Crop Science, Chinese Academy of Agricultural Sciences;
[0075] The preservation numbers of the National Resource Bank of Century near-isogenic materials in the United States are: Lox-1: WDD01483, Lox-3: WDD01485;
[0105] Example 2: Preparation, detection and application of monoclonal antibodies
[0106] 2.1 Detection antibody preparation
[0107] 2.1.1 Monoclonal antibody preparation and purification
[0108] 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.
[0109] Ascitestiter 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 antibodies in ascites were 1:320,000, 1:640,00...
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technical field [0001] 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: CGMCC 5121, CGMCC 5122 and CGMCC 5123, and also involve the application of the monoclonal antibodies secreted by each of them in the detection of soybean lipoxygenase isozymes by double-antibody sandwich enzyme-linked immunosorbent assay (DAS-ELISA), It belongs to the technical field of biological detection. Background technique [0002] Soybean contains three lipoxygenase (Lox for short) isoenzymes namely Lox1, Lox2 and Lox3, which can specifically catalyze polyunsaturated fatty acids with cis, cis-1, 4-pentadiene structure through molecular addition Oxygen, forming hydroperoxide derivatives with conjugated double bond...
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Patent Type & Authority Patents(China)
IPC IPC(8): C12N5/20C07K16/40G01N33/577G01N33/573
Inventor 张孟臣马志民蒋春志邸锐杨春燕赵双进秦君闫龙刘兵强赵青松唐晓东史晓蕾李亚璞
Owner INST OF CEREAL & OIL CROPS HEBEI ACAD OF AGRI & FORESTRY SCI
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