Vibrio outer membrane protein subunit adjuvant vaccine and large-scale preparation technique
A technology of outer membrane protein and subunit, which is applied in antibacterial drugs, medical preparations containing active ingredients, antibody medical ingredients, etc., and can solve problems such as the lack of subunit vaccines for the prevention and control of marine fish disease immunity, and the late start of development.
Inactive Publication Date: 2017-07-07
PEARL RIVER FISHERY RES INST CHINESE ACAD OF FISHERY SCI
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Problems solved by technology
Due to the late start of the development of aquatic vaccines in my country, the subunit vaccines used in the prevention and control of diseases in some aquatic products are basically still in the laboratory scale. At present, there are no subunit vaccines that can be produced on a large scale and applied to the prevention and control of diseases in seawater fish.
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[0021] The technical solutions of the present invention will be described in detail below through specific examples, but the scope of the present invention is not limited by these examples.
[0022] This embodiment provides a large-scale production technology of Vibrio outer membrane protein subunit adjuvant vaccine for immune prevention and treatment of seawater fish vibriosis, which specifically includes the following steps:
[0023] (1) Primary seed preparation
[0024] The engineered bacteria were inoculated on nutrient agar, 30°C, cultivated for 24 hours, picked bacterial moss and inoculated 9 bottles of 300ml nutrient broth / bottle, 30°C, 180r / min, cultivated for 10 hours, and obtained 2.7L shake flask seed liquid. The Escherichia coli DH5α (pBVOmpK) selected in this example is a recombinant bacterium prepared by our laboratory. Competent Escherichia coli Dh5 was purchased from Takara.
[0025] (2) Secondary seed preparation
[0026] 2.7L shake flask seed liquid was in...
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Abstract
The invention relates to a vibrio harveyi outer membrane protein subunit adjuvant vaccine and particularly relates to a vibrio harveyi outer membrane protein subunit adjuvant vaccine and a large-scale preparation technique thereof. The vibrio harveyi outer membrane protein subunit adjuvant vaccine contains a vibrio harveyi outer membrane protein Ompk and a Montanide ISA 763A(763A) adjuvant. The invention further provides a large-scale preparation method of the vibrio harveyi outer membrane protein subunit adjuvant vaccine. The preparation method is characterized by comprising the steps of carrying out high-density culture and induction expression on escherichia coli (engineering bacteria) carrying and containing vibrio outer membrane protein Ompk gene plasmids by virtue of a 500L fermentation tank, extracting, purifying to obtain the vibrio harveyi outer membrane protein Ompk, emulsifying the vibrio harveyi outer membrane protein Ompk together with the Montanide ISA 763A(763A) adjuvant in a certain proportion, and carrying out sterile split charging, so as to prepare a vibrio outer membrane protein Ompk adjuvant vaccine finished product. The immune protective rates of the vaccine to Saddletail grouper and crucians reach above 60%.
Description
technical field [0001] The invention belongs to the technical field of immune prevention and control of aquaculture diseases, and in particular relates to a seawater vibrio ichthyosis outer membrane protein subunit adjuvant vaccine and a large-scale preparation technology. Background technique [0002] Pathogenic Vibrio is the most important bacterial pathogen of marine animals, and the disease caused by it is often called "vibriosis". Vibrio harveyi is one of the important pathogens of bacterial diseases in marine fish farming. While developing various inactivated vaccines, researchers are also developing outer membrane protein subunit vaccines to prevent and control marine fish farming diseases. Due to the late start of the development of aquatic vaccines in my country, the subunit vaccines used in the prevention and control of diseases in some aquatic products are basically still in the laboratory scale. At present, there are no subunit vaccines that can be produced on a...
Claims
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IPC IPC(8): A61K39/106A61K39/39A61P31/04
CPCA61K39/107A61K39/39A61K2039/55566
Inventor 陶家发孙承文赖迎迢江小燕罗霞赵长臣巩华刘春花黄志斌
Owner PEARL RIVER FISHERY RES INST CHINESE ACAD OF FISHERY SCI