Eimeria acervulina nano-subunit vaccine, preparation method and application thereof

A technology of Eimeria and heap-type, which is applied in the field of Eimeria heap-type nano-subunit vaccine and its preparation, can solve the problems such as the application research report of PLGA nano-subunit vaccine, and achieve the effect of immune protection The effect of enhancement and embedding rate improvement

Active Publication Date: 2019-12-13
NANJING AGRICULTURAL UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

There is no research report on the application

Method used

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  • Eimeria acervulina nano-subunit vaccine, preparation method and application thereof
  • Eimeria acervulina nano-subunit vaccine, preparation method and application thereof
  • Eimeria acervulina nano-subunit vaccine, preparation method and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0029] Example 1. Expression and purification of Eimeria acervulina recombinant protein EaMIC3

[0030] 1. Preparation of genetically engineered bacteria

[0031] The E. acervulina pET-32a-EaMIC3 ORF (Zhang Zhenchao, the role of Eimeria acerciformis microwire protein in the specificity of invasion site. Nanjing Agricultural University PhD dissertation, 2016) was transformed into E.coli BL21 receptor. Cells, and get bacteria containing pET-32a-EaMIC3 ORF recombinant expression plasmid.

[0032] 2. Expression and purification of Eimeria acervulina recombinant protein EmMIC3

[0033] Bacteria containing pET-32a-EaMIC3 ORF recombinant expression plasmid were inoculated into LB liquid medium at a volume ratio of 1:100, and cultivated to OD at 37℃, 200r / min 600 When it is 0.4-0.6, add IPTG with a final concentration of 1mmol / L to induce expression. Purify the EMIC3 recombinant protein in accordance with the GE protein purification kit instructions, and subject the collected protein samples...

Embodiment 2

[0036] Example 2. Immune protection test of Eimeria acervulina recombinant protein subunit vaccine and nano subunit vaccine 1. Experiment design

[0037] 0-day-old Hailan white chicks are raised in a strictly disinfected, coccidia-free environment, and they are free to drink and eat. The 14-day-old chickens were weighed one by one, and overweight and underweight individuals were eliminated. Randomly grouped and adjusted the average weight between the test groups so that the average weight was close to the same, 15 birds in each group; recombinant proteins EaMIC3 and pET- were used respectively. The PLGA-EaMIC3 and PLGA-pET-32a-tagged proteins are nano subunit vaccines prepared by encapsulating recombinant proteins with 32a-tagged protein, PLGA, and PLGA nanomaterials to immunize chickens by intramuscular injection into the leg. The immunization dose is 100μg, and it is administered at 21 days of age. Booster immunity, oral infection of fresh E.acervulina sporulated oocysts 10×10 ...

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Abstract

The present invention discloses an eimeria acervulina nano-subunit vaccine, a preparation method and an application thereof. The eimeria acervulina nano-subunit vaccine is a nano-particle formed by PLGA encapsulating a recombinant protein EaMIC3, and the recombinant protein EaIMIC3 is an eimeria acervulina microneme protein 3 and has an amino acid sequence shown as SEQ ID NO.1. The recombinant protein EaMIC3 is coated by the nano-material PLGA to form a brand-new vaccine form and to obtain the eimeria acervulina PLGA nano-subunit vaccine with a relatively high immune protection effect.

Description

Technical field [0001] The invention relates to the technical field of biological veterinary medicines, and relates to an Eimeria acervulina nano subunit vaccine and a preparation method and application thereof. Background technique [0002] Chicken coccidiosis is an intestinal disease caused by protozoan parasites of the genus Eimeria. It is one of the most important poultry diseases in the world. The incidence rate of chicken coccidiosis is 50-70%, the mortality rate is 20-30%, and it is as high as 80% in severe cases. Chicken coccidiosis can cause extensive damage to the intestines of chickens, weight loss of infected chickens, lower feed conversion rate, lower egg production rate of laying hens, and economic losses of more than 3 billion US dollars to the poultry industry every year. At present, the main method to control chicken coccidiosis is to add precise amounts of anticoccidiosis drugs to the feed. However, long-term use of anticoccidial drugs has led to the continuou...

Claims

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Application Information

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IPC IPC(8): A61K39/012A61K9/51A61K47/34A61P33/02C12N15/70C07K14/455
CPCA61K39/012A61K9/5153A61P33/02C07K14/455C12N15/70Y02A50/30
Inventor 李祥瑞严若峰宋小凯徐立新黄剑梅
Owner NANJING AGRICULTURAL UNIVERSITY
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