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Nanoliposome antivenom active enzyme A2 and enteropathogen specific composite IgY and application thereof

An enteric pathogen, specific technology, applied in the application, anti-enzyme immunoglobulin, hydrolase, etc., can solve the problems of rising feed raw material prices, low feed conversion rate, and disrupting intestinal flora balance.

Active Publication Date: 2015-06-17
SHENZHEN JASON INTELLIGENT BIOTECH CO LIMLTED PRC
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0003] (1) The feed conversion rate is not high: a snake venom active enzyme A2 (including lipoprotein phospholipase A2 and phospholipase A2) secreted by livestock, poultry and aquatic animals, these enzymes are equivalent to the "roundworms" in the human body, and will prevent The absorption of nutrients by the digestive system results in a low feed conversion rate; it not only restricts the operating efficiency of breeding enterprises, but also causes long-term contradictions between food supply and demand, causing the price of feed raw materials to continue to rise
[0004] (2) The outbreak of infectious diseases is on the verge of multiplying: diarrhea in autumn and winter has become an epidemic, and highly pathogenic diseases such as blue ear disease and foot-and-mouth disease often cause large-scale deaths of pigs and cattle, resulting in disastrous consequences. Infectious diseases of fish and shrimp Outbreaks also bring devastating blows to aquaculture farms from time to time; thus, seriously affect the economic benefits of the aquaculture industry
[0005] (3) Abuse of antibiotics and antiviral chemicals: Since antibiotics are broad-spectrum bactericidal, they kill harmful bacteria in animals as well as beneficial bacteria, which breaks the balance of intestinal flora , reduces intestinal immunity
However, the existing antiviral drugs only have a certain degree of interference or influence on a certain stage of a certain virus replication process, and have no effect on viruses that have entered the cell.
The harsh reality is that so far, antiviral drugs that can inactivate viruses without harming host cells have not been successfully developed in the world

Method used

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  • Nanoliposome antivenom active enzyme A2 and enteropathogen specific composite IgY and application thereof
  • Nanoliposome antivenom active enzyme A2 and enteropathogen specific composite IgY and application thereof
  • Nanoliposome antivenom active enzyme A2 and enteropathogen specific composite IgY and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0224] Preparation of anti-lipoprotein phospholipase A2 and porcine enteropathogen-specific complex IgY:

[0225] (1) Preparation of specific compound IgY against lipoprotein phospholipase A2 and porcine enteropathogenic bacteria

[0226] 1. Prepare the expressed protein antigen component of lipoprotein phospholipase A2, the specific steps are as follows:

[0227] The lipoprotein phospholipase A2 gene was subcloned from the eukaryotic expression vector pCGN-PLD1 into the shuttle plasmid pAdTrack-CMV with a green fluorescent marker protein; then homologous recombination was performed in Escherichia coli BJ5183 together with the adenovirus backbone vector; positive After the recombinant was linearized by Pac I, it was transfected into the virus assembly cell line 293 cells, and the specific lipoprotein phospholipase A2 recombinant adenovirus expression protein was successfully constructed; the virus particles were used to infect the pheochromocytoma cell PC12 cells, and highly e...

Embodiment 2

[0271] Preparation of anti-lipoprotein phospholipase A2 and porcine enteric pathogen-specific complex IgY pre-ingredients:

[0272] Using the prepared nano-sized anti-lipoprotein phospholipase A2 and porcine intestinal pathogen-specific compound IgY1050g as the main raw materials to make a high-efficiency and safe drug that can improve the nutrient absorption function of piglets and prevent and treat porcine transmissible gastroenteritis and epidemic diarrhea green pre-mixed ingredients.

[0273]

[0274] Add both ingredients to a blender and mix well. Then use a packaging machine to pack in 500g, and then pack. Leave the factory after passing the sampling inspection.

Embodiment 3

[0276] Preparation of anti-phospholipase A2-specific IgY:

[0277] 1. Prepare the expression protein antigen component of phospholipase A2, the specific steps are as follows:

[0278]The phospholipase A2 gene was subcloned from the eukaryotic expression vector pCGN-PLD1 into the shuttle plasmid pAdTrack-CMV with a green fluorescent marker protein; then homologous recombination was carried out in Escherichia coli BJ5183 together with the adenovirus backbone vector; positive recombinants After linearization by Pac I, it was transfected into the virus assembly cell line 293 cells, and the specific phospholipase A2 recombinant adenovirus expression protein was successfully constructed; the virus particles were used to infect pheochromocytoma PC12 cells, and the phospholipase A2 recombinant was highly expressed Adenovirus-expressed protein. The identified phospholipase A2 recombinant adenovirus expressed protein was purified by cesium chloride gradient centrifugation, and then mas...

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Abstract

The invention discloses a method for preparing nanoliposome antivenom active enzyme A2 (comprising lipoprotein phospholipase A2 and phospholipase A2) and enteropathogen specific composite IgY and antivenom active enzyme A2 (comprising lipoprotein phospholipase A2 and phospholipase A2) specific composite IgY. The method comprises the following steps of: (1) reconstructing the lipoprotein phospholipase A2 and the phospholipase A2 expression protein by using genetic engineering technology; (2) preparing domestic aquatic animal main enteropathogen composite antigen; (3) preparing the antivenom active enzyme A2 and enteropathogen specific composite IgY and the antivenom active enzyme A2 specific IgY; (4) preparing a rough product of lipoprotein phospholipase A2 and domestic aquatic animal enteropathogen specific composite IgY; and (5) nanorizing and performing liposome processing on the antivenom active enzyme A2 and enteropathogen specific composite IgY and the antivenom active enzyme A2 specific IgY. A feed or feed additive for domestic aquatic animals prepared by the specific composite IgY or IgY can enhance the health of the animal intestines, suppress special phospholipase which can affect the nutrient absorption and improve the conversion efficiency of the feed.

Description

technical field [0001] The present invention relates to a feed and feed additive for livestock, poultry and aquatic animals, in particular to a nano-liposome anti-venom active enzyme A2 and enteropathogen-specific composite IgY and anti-venom active enzyme A2-specific IgY. The snake venom active enzyme can be lipoprotein phospholipase A2 or phospholipase A2, and the invention also provides the application of this IgY in livestock, poultry and aquatic animal feed and feed additive. Background technique [0002] Current problems in China's aquaculture industry: [0003] (1) The feed conversion rate is not high: a snake venom active enzyme A2 (including lipoprotein phospholipase A2 and phospholipase A2) secreted by livestock, poultry and aquatic animals, these enzymes are equivalent to "roundworms" in the human body, and will prevent The absorption of nutrients by the digestive system leads to a low feed conversion rate; it not only restricts the operating efficiency of breedi...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C07K16/40C07K16/02C07K1/18C07K1/16C12N15/55C12N15/861C12N9/16A23K1/16A23K1/18A23K20/10
Inventor 包晟杨荣鉴汪玲张善钧
Owner SHENZHEN JASON INTELLIGENT BIOTECH CO LIMLTED PRC