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Hybridoma of DON (Deoxynivalenol) monoclonal antibody as well as preparation method and application thereof

A technology of deoxynivalenol and monoclonal antibody, which is applied in the field of bioengineering and can solve difficult and demanding problems

Inactive Publication Date: 2011-05-25
中国疾病预防控制中心营养与食品安全所 +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, in this study, the same protein and DON conjugate (DON-BSA) were used in the detection of the immunized animals and the monoclonal antibody, and the method of preparing anti-DON monoclonal antibody by the carrier exclusion method has not been reported at home and abroad.
Because only one complete antigen needs to be synthesized, although this method is economical, simple, and time-saving, it is technically more demanding and difficult than the method of using different complete antigens for immunization and detection.

Method used

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  • Hybridoma of DON (Deoxynivalenol) monoclonal antibody as well as preparation method and application thereof
  • Hybridoma of DON (Deoxynivalenol) monoclonal antibody as well as preparation method and application thereof
  • Hybridoma of DON (Deoxynivalenol) monoclonal antibody as well as preparation method and application thereof

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Experimental program
Comparison scheme
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Embodiment Construction

[0028] (1) Preparation of anti-DON monoclonal antibody

[0029] 1. Animal immunization

[0030] BSA and DON are coupled with 1,3 dicyclyl diimide (DCC) and N-hydroxysuccinimide (NHS) respectively, and the two are connected by glutaric acid to form a DON-BSA conjugate, and DON and The molar ratio of BSA was 50:1. The method has high coupling efficiency, stable coupled antigen, good solubility and strong immunogenicity.

[0031] Prepare the DON-BSA conjugate into a 1mg / ml solution, adopt a small-dose and long-term immunization scheme, and use 100μg DON-BSA for the first immunization of 6-8 week-old female Balb / c mice (body weight 18-20g) , adding an equal amount of complete Freund's adjuvant (Sigma company product), intraperitoneal injection. After 2 weeks, 100 μg of DON-BSA was added and an equal amount of incomplete Freund's adjuvant (product of Sigma Company) was added for intraperitoneal injection. After another 2 weeks, 100 μg of DON-BSA was added into the same amount o...

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Abstract

The invention relates to the biological engineering field, in particular to hybridoma of a DON (Deoxynivalenol) monoclonal antibody as well as a preparation method and application thereof. Through cell fusion, an immunizing antigen and a detection antigen adopt DON-BSA (Bovine Serum Albumin) to obtain and store hybridoma secreting the DON monoclonal antibody with high efficiency, and the antibody has high sensitivity, strong specificity and appetency with DON and low cross reaction with other toxins. The invention lays the foundation for researching DON test reagents.

Description

Technical field: [0001] The invention relates to the field of bioengineering, in particular to a hybridoma cell anti-deoxynivalenol (DON) monoclonal antibody and its preparation method and application. Background technique: [0002] Among the many pollution factors of agricultural products in my country, mycotoxins are an important class of natural pollutants. The food-borne diseases and food trade disputes caused by them have always been the focus of global attention. Among them, Deoxynivalenol (DON) is the most toxic mycotoxin, most closely related to human health, most polluting to crops, and most widely distributed. Deoxynivalenol (DON) is not only the main toxin that contaminates scab wheat, but also contaminates corn, barley, sorghum and other grains. People and animals can be poisoned after eating food and feed contaminated by this toxin, producing DON Fusarium is also the main pathogenic bacteria that causes rice bakanae disease, vegetables (such as zucchini zucchin...

Claims

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Application Information

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IPC IPC(8): C12N5/20C07K16/14G01N33/577G01N33/569C12R1/91
Inventor 李凤琴赵丽王伟
Owner 中国疾病预防控制中心营养与食品安全所
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