E.maxima nanometer subunit vaccine and preparation method and application thereof

A Eimeria gigantic technology, applied in the field of Eimeria gigantic nano-subunit vaccines and its preparation, achieves improved embedding rate, improved immune protection effect, and excellent protection effect

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

AI Technical Summary

Problems solved by technology

It has been reported that E.tenella microneptin 3 (EtMIC3) is the key molecule that determines the parasitism of E. tenella in the cecum, but the key molecule that determines the parasitism site of E. maxima has not been reported yet.

Method used

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  • E.maxima nanometer subunit vaccine and preparation method and application thereof
  • E.maxima nanometer subunit vaccine and preparation method and application thereof
  • E.maxima nanometer subunit vaccine and preparation method and application thereof

Examples

Experimental program
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Effect test

Embodiment 1

[0029]Example 1. Expression and purification of Eimeria maxima EmMIC3 recombinant protein

[0030] 1. Preparation of genetically engineered bacteria

[0031] Immunoprotective properties of E.maxima pET-32a(+)-MIC3 (Huang Jingwei, Eimeria maxima, four nemelin proteins and their binding ability to chicken intestinal epithelial cells. Doctoral dissertation of Nanjing Agricultural University, 2016) The recombinant expression plasmid was transformed into E.coli BL21 competent cells to obtain bacteria containing the recombinant expression plasmid of pET-32a(+)-MIC3.

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

[0033] Inoculate the bacteria containing the pET-32a(+)-MIC3 recombinant expression plasmid in LB liquid medium at a volume ratio of 1:100, and cultivate to OD at 37°C and 200r / min 600 When it is 0.4-0.6, add IPTG with a final concentration of 1mmol / L to induce expression. The EmMIC3 recombinant protein was purified according to the...

Embodiment 2

[0036] Example 2. Immunoprotective Detection of Eimeria maxima Recombinant Protein Subunit Vaccine and Nano Subunit Vaccine

[0037] 1. Experimental design

[0038] The 0-day-old Hy-Line white chicks were raised in a strictly sterilized, coccidia-free environment, and had free access to water and food. The 14-day-old chickens were weighed one by one, and the individuals who were overweight and underweight were eliminated, and the average body weight between the test groups was randomly divided into groups to make the average body weight close to the same, with 20 birds in each group; the recombinant proteins EmMIC3, pET- 32a-tagged protein, PLGA, and PLGA-EmMIC3 and PLGA-pET-32a-tagged protein, which are prepared by encapsulating recombinant protein with PLGA nanomaterials, were used to immunize chickens by intramuscular injection of 100 μg in the leg muscle, at 21 days of age Booster immunization, orally infect fresh sporulated oocysts of E.maxima 10×10 at the age of 28 days...

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Abstract

The invention relates to an E.maxima recombinant protein subunit vaccine EmMIC3 and a nanometer subunit vaccine PLGA-EmMIC 3, and belongs to the technical field of biological veterinary medicines. Thenanometer subunit vaccine is prepared by wrapping recombinant protein subunit vaccine EmMIC3 with PLGA nanometer material. Animal immune protective tests prove that the recombinant protein subunit vaccine EmMIC3 and the nanometer subunit vaccine PLGA-EmMIC3 can effectively resist the infection of the E.maxima. The invention uses nanometer material PLGA to wrap the recombinant protein EmMIC3 to form a brand-new vaccine form, which is different from the single EmMIC3 recombinant protein, and the two components are different. After the recombinant protein EmMIC3 and the nanometer material PLGA are coated, the immune protection effect is obviously improved, and the protection effect is improved from general level to excellent level.

Description

technical field [0001] The invention relates to the technical field of biological veterinary medicine, and relates to a nano-subunit vaccine of Eimeria maxima and a preparation method and application thereof. Background technique [0002] Chicken coccidiosis is a globally prevalent parasitic disease that seriously endangers the poultry industry. Its morbidity rate is 50-70%, mortality rate is 20-30%, can reach more than 80% when serious. The production efficiency of resistant chickens is greatly reduced, causing economic losses of more than 3 billion US dollars per year. At present, chicken coccidiosis is mainly controlled by drugs. However, with the continuous emergence of drug-resistant strains, the preventive effect of drugs has been significantly reduced, and chicken coccidia drug resistance and anticoccidial drug residues during the use of drugs have seriously affected the safety of animal-sourced food. Therefore, efforts are being made to find alternatives to coccid...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K39/012A61K9/51A61K47/34A61P33/02C12N15/70C07K14/455
CPCA61K39/012A61K9/5153A61P33/02C07K14/455C12N15/70Y02A50/30
Inventor 李祥瑞宋小凯严若峰徐立新黄剑梅
Owner NANJING AGRICULTURAL UNIVERSITY
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