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Mononal antibody against avermectin, hybridoma cell line secreting the same and its preparation method

A hybridoma cell line, anti-avermectin single technology, applied in the field of immunology, can solve the application prospect discount of immunodetection method, polyclonal antibody does not have unlimited supply, polyclonal antibody homogeneity, specificity is inferior to monoclonal antibody. Cloning antibodies, etc.

Inactive Publication Date: 2006-02-15
NORTHEAST AGRICULTURAL UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

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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  • Mononal antibody against avermectin, hybridoma cell line secreting the same and its preparation method
  • Mononal antibody against avermectin, hybridoma cell line secreting the same and its preparation method
  • Mononal antibody against avermectin, hybridoma cell line secreting the same and its preparation method

Examples

Experimental program
Comparison scheme
Effect test

Embodiment

[0039] Materials and Instruments

[0040] 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

fetal bovine serum

Tianjin

liquid paraffin oil

Beijing Reagent Company

bovine serum albumin

Huamei Bioengineering Company

ovalbumin

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

OVA

SIGMA

tert-Butyldimethylc...

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PUM

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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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Application Information

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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
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