Mononal antibody against avermectin, hybridoma cell line secreting the same and its preparation method
A hybridoma cell line, anti-abamectin single technology, applied in the field of immunology, can solve the discount of application prospects of immunoassay methods, polyclonal antibody does not have unlimited supply, polyclonal antibody uniformity, specificity is not as good as single Issues such as cloning antibodies
Inactive Publication Date: 2007-06-20
NORTHEAST AGRICULTURAL UNIVERSITY
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Problems solved by technology
Although the detection method is simpler than before, the antibodies used in the established immunoassay method are still polyclonal antibodies. Antibodies have the characteristics of unlimited supply, so the application prospect of this immunoassay method is greatly reduced
Method used
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Embodiment
[0040] Materials and Instruments
[0041] Main reagents and purchase sources
[0042] Polyethylene glycol (PEG)
SIGMA
Freund's complete adjuvant (FCA)
Beijing Xinjingke Biological Co., Ltd.
Freund's incomplete adjuvant (FICA)
Beijing Xinjingke Biological Co., Ltd.
Dimethylsulfoxide (DMSO)
Harbin Yishida Biological Engineering Co., Ltd.
HRP-labeled goat anti-mouse IgG
Beijing Zhongshan Jinqiao Biotechnology Co., Ltd.
HAT and HT
GIBCO
DMEM
GIBCO
Tianjin
liquid paraffin oil
Beijing Reagent Company
Huamei Bioengineering Company
Huamei Bioengineering Company
IgG Antibody Subclass Kit
Southern Biotech
PEG(1450)
SIGMA
AVM
Provided by China Agricultural University Microbiology Factory
IVM
Provided by China Agricultural University Microbiology Factory
BSA
SIGMA
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Abstract
The invention provides a novel Avermectin resistant monoclonal antibody, hybridomas cell line excreting it and its preparing process, which comprises synthesizing artificial antigen AVM-BSA, immuning BALB / c mouse for five times, carrying out cell fusion to mouse spleen cells and SP2 / O marrow tumor cells, using PEG1450 as fusion agent to screen positive fused cells, subcloning five times with limited dilution method, obtaining hybridomas cell lines 2F2 specifically secreting AVM antibody, whose micro-organism Deposit number is CGMCC NO.1413, injecting BALB / c mouse's abdominal cavity with the hybridomas cell, gathering ascitic fluid, centrifuging, gathering supernatant fluid, purifying and identifying to obtain the Avermectin resistant monoclonal antibody.
Description
technical field [0001] The invention relates to a monoclonal antibody, in particular to an anti-abamectin monoclonal antibody, a hybridoma cell line secreting it and a preparation method, belonging to the field of immunology. Background technique [0002] Avermectin (AVM) is currently the most widely used anti-endoparasite drug in veterinary clinics, and it is also an agricultural insecticide widely used in agricultural production. Since the first discovery in 1970 by scientists of Merk Sharp and research laboratory in the United States in the screening of soil actinomycete samples from Kitasato Research Institute in Japan, AVMs have been widely used worldwide (Egerton J.E, et al, 1979). As an antiparasitic drug, AVMs has a novel chemical structure and a unique mechanism of action. Due to its specific anthelmintic effect and high safety, it is regarded as the most excellent and most widely used antiparasitic drug for veterinary use with high efficiency and low toxicity. Pa...
Claims
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IPC IPC(8): C12N5/18C07K16/00C12P21/08G01N33/53
Inventor 李术徐世文熊永忠
Owner NORTHEAST AGRICULTURAL UNIVERSITY