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Vibrio harveyi recombined outer-membrane protein Ompk microspheres vaccine and preparation method thereof

A technology of Vibrio harveyi and microsphere vaccines, which can be applied to antibacterial drugs, pharmaceutical formulas, medical preparations with non-active ingredients, etc., can solve the problems of difficult operability of fish vaccines, and achieve large-scale use, Antigen content is high, the effect of preventing vibriosis

Inactive Publication Date: 2009-08-19
PEARL RIVER FISHERY RES INST CHINESE ACAD OF FISHERY SCI
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Problems solved by technology

[0003] The purpose of the present invention is to provide a vibrio microsphere vaccine that can be taken orally and its preparation method for the shortcomings of the existing vibriosis vaccine injection or immersion immunization, so as to solve the problem of difficult operability of fish vaccines in production practice

Method used

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  • Vibrio harveyi recombined outer-membrane protein Ompk microspheres vaccine and preparation method thereof

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manufacture example

[0016] Manufacturing example: preparation of Vibrio harveyi recombinant outer membrane protein Ompk microsphere vaccine

[0017] Inoculate the Escherichia coli engineering strain carrying the pBV220 prokaryotic expression plasmid of the Ompk gene on an LB plate containing 50 μg / ml ampicillin, and culture it at 28°C for 18-20h; pick a single colony and inoculate it in the LB liquid medium, culture it at 30°C for 12h, and then Transfer to LB liquid medium at a ratio of 1:100, culture at 30°C for 6 hours, raise the temperature sharply to 42°C, induce culture for 7 hours, collect the induced cells by centrifugation, ultrasonically break, and extract the inclusion bodies. After washing with urea solution, dissolve with 8M urea solution, dialysis and concentrate through tangential flow ultrafiltration system to obtain recombinant protein with higher purity ( figure 1 ). The concentration of the recombinant protein was measured, and after filter sterilization, it was stored at 4°C f...

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Abstract

The invention discloses a microsphere vaccine and a preparation method thereof. The vaccine is the microsphere vaccine which is prepared from recombinant outer-membrane protein Ompk of vibrio harveyi as a common mariculture fish pathogen, as well as biodegradable polymer materials through emulsification and drying. The microsphere vaccine is simple in preparation process, convenient for factory production, stable and reliable in production, safe and convenient to use, and the product yield reaches more than 80 percent. The microsphere vaccine has the advantages of high antigen content, long sustained release time, few toxic-side effects, convenience for large-scale application and the like. The prepared microsphere vaccine which is orally taken for immunization can stimulate an immune system to produce immune response in a long period of time, has high relative protection ratio to tested fishes, and can effectively prevent mariculture fish vibriosis.

Description

technical field [0001] The present invention relates to a microsphere vaccine. The invention also relates to a preparation method of the microsphere vaccine. Background technique [0002] Vibrio is a class of Gram-negative, polar-flagellated, motile, non-spore-forming brevibacteria or Vibrio bacteria. It is one of the most common bacterial groups in the marine environment, and some species are pathogenic bacteria of marine organisms. Vibrio harveyi (Vh) is one of the common important pathogenic bacteria, which can infect a variety of farmed animals and cause huge economic losses. In recent years, marine Vibrio vaccines have played an increasingly important role in the prevention and treatment of vibriosis. There are four main ways to immunize fish vaccines: injection method, spray method, soaking method and oral method. The injection method has the advantages of less dosage and good immune effect, but it is time-consuming, laborious, inconvenient to operate, and easy to da...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K39/106A61K9/16A61K47/36A61P31/04
Inventor 任燕吴淑勤李宁求常藕琴石存斌
Owner PEARL RIVER FISHERY RES INST CHINESE ACAD OF FISHERY SCI
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