Anti-Vibrio anguillarum yolk antibody, and preparation method, titer detection method and application thereof
A technology of yolk antibody and Vibrio anguillarum, which is applied in the direction of antibodies, antibacterial drugs, antibacterial immunoglobulin, etc., can solve the problem of no anti-Vibrio anguillarum yolk antibody, etc., achieve cheap acquisition, strong inhibitory effect, and simple preparation method easy effect
Inactive Publication Date: 2012-07-04
NINGBO UNIV
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At present, yolk antibody has been widely concerned and applied in animal husbandry, but the research and development in the aquaculture industry is still in the basic stage. At present, the patents applied for publication and authorization in aquaculture include anti-prawn virus disease, anti-Cryptosporidium bayeri parahaemolyticus disease and anti-Aeromonas hydrophila yolk antibody, but there is no relevant research report on anti-Vibrio anguillarum yolk antibody and its preparation method and application
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[0021] The present invention will be further described in detail below in conjunction with the accompanying drawings and embodiments.
1. Specific examples
[0022] An anti-vibrio anguillarum yolk antibody of the present invention is prepared by preparing vibrio anguillarum inactivated antigen immunized eggs, collecting highly immune eggs for separation and purification, and performing protein concentration determination and SDS-PAGE detection of protein bands to determine the purity, and indirect ELISA detection Antibody titer, antibacterial experiment in vitro to determine the effect, and finally obtain high-yield, high-efficiency, high-purity egg yolk antibody. The preparation process is as follows:
[0023] (1) Preparation of Vibrio anguillarum antigen and inactivation
[0024] Inoculate Vibrio anguillaris on LB solid agar medium, and culture it upside down at 28°C for 18 hours, then pick a single colony and inoculate it in LB liquid medium, shake the shaker at 28°C and 1...
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Abstract
The invention discloses an anti-Vibrio anguillarum yolk antibody, and the preparation method, the titer detection method and the application thereof. The invention is characterized in that the anti-Vibrio anguillarum yolk antibody is obtained by immunizing laying hens with Vibrio anguillarum as antigen, collecting highly-immunized eggs and performing separation and purification of yolk of the immunized eggs. The method comprises a step of preparing a Vibrio anguillarum antigen and inactivating the antigen, a step of performing basic immunization once in laying hens and secondary immunization for two to three times and collecting eggs, and a step of performing centrifugal separation, salting-out and dialysis to obtain the final product. The titer of the anti-Vibrio anguillarum yolk antibody is determined by indirect enzyme-linked immunosorbent assay. The anti-Vibrio anguillarum yolk antibody can be used for prevention and treatment of diseases caused by Vibrio anguillarum infection and can also be used for development of Vibrio anguillarum test kits and immunoreaction and diagnosis in immunology. The invention has the advantages that the yolk antibody is higher in purity and has a significant inhibitory effect on Vibrio anguillarum; the extraction of highly-immunized yolk is cost-efficient and convenient, and is adapted to low-cost and mass production; and the preparation method is simple and feasible.
Description
technical field [0001] The invention relates to the field of preparation technology and application of biological products, in particular to an anti-vibrio anguillarum yolk antibody and its preparation method and application. Background technique [0002] Vibrio anguillarum ( Vibrio anguillarum ) is a highly prevalent aquatic animal pathogen, which can cause disease to both seawater and freshwater animals, and has brought huge losses to farmers all year round. According to reports, Vibrio anguillaris can infect flounder, sea bass, salmon, yellow croaker, rainbow trout, eel, sweetfish, cod, turbot, etc.; Shrimp and shellfish have a strong stimulating effect on the immune system and are the most harmful bacteria in Vibrio. [0003] At present, there are the following methods for the prevention and treatment of diseases caused by Vibrio anguillarum: antibiotics, Chinese herbal medicines, inactivated vaccines, and monoclonal antibodies. Antibiotics and chemical drugs are easy...
Claims
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IPC IPC(8): C07K16/12C07K16/02G01N33/569A61K39/40A61P31/04
Inventor 李登峰周永强刘联国郭安南彭娇陈炯顾晓英
Owner NINGBO UNIV
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