Compound preparation with egg yolk antibodies for resisting dairy cow mastitis and method for preparing compound preparation

A technology of egg yolk antibody and compound preparation, which is applied in the direction of antibodies, antibacterial drugs, drug combinations, etc., can solve the problems of mixed infection that cannot be well controlled and limits the application of egg yolk antibody, and achieves high purity, low preparation cost, and specificity. strong effect

Inactive Publication Date: 2017-05-17
DALIAN UNIV OF TECH +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, most of the antibody preparations currently used are single-pathogen antibodies, which cannot control mixed infections well
On the other hand, the application of egg yolk antibody to prevent and treat diseases also has unavoidable disadvantages, such as the possibility of carrying the pathogen of egg infectious diseases, which limits the application of egg yolk antibody in the poultry industry

Method used

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  • Compound preparation with egg yolk antibodies for resisting dairy cow mastitis and method for preparing compound preparation
  • Compound preparation with egg yolk antibodies for resisting dairy cow mastitis and method for preparing compound preparation
  • Compound preparation with egg yolk antibodies for resisting dairy cow mastitis and method for preparing compound preparation

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0035] Example 1 Preparation of specific egg yolk antibody against main pathogenic bacteria of dairy cow mastitis

[0036] Take the preparation of anti-Staphylococcus aureus antibody as an example.

[0037] (1) Preparation of Staphylococcus aureus antigen

[0038] Staphylococcus aureus was cultured upside down on TSA solid agar medium at 37°C for 24 hours, then a single colony was picked and inoculated in nutrient broth liquid medium, and cultured on a shaker at 37°C and 140 rpm for 12 hours, then cultured to a bacterial concentration of 10 8CFU / mL, and then centrifuged to obtain the precipitate and supernatant containing Staphylococcus aureus, the precipitate was mixed with sterile PBS to obtain 10 9 CFU / mL bacterial suspension of Staphylococcus aureus.

[0039] (2) Inactivation of Staphylococcus aureus antigen

[0040] Add formaldehyde to the bacterial suspension so that the final volume concentration of formaldehyde is 0.5%. After inactivation on a shaking table at 37°C...

Embodiment 2

[0048] Example 2 Titer Detection of Egg Yolk Antibody

[0049] The titer was detected by enzyme-linked immunosorbent assay, with 10 9 Pathogen-coated ELISA plates with a concentration of CFU / mL bacteria, 100 μL per well, coated overnight at 4 °C, swung the plate, washed 3 times with washing solution, added blocking solution containing 1% BSA and bathed in water at 37 °C for 2 h. After the water bath, shake the plate, wash 3 times, and then add 100 μL of egg yolk antibody (or crude extract of egg yolk antibody) diluted with conjugate diluent containing 1‰BSA to each well, and set negative control and blank control at the same time, 37°C Water bath 2h. After the water bath, wash 3 times, add 100 μL of peroxidase-labeled rabbit anti-chicken antibody (IgG-HRP) diluted in conjugate diluent to each well, and bathe in water at 37°C for 1 hour. Wash 5 times after the reaction, add 100 μL of TMB chromogenic solution to each well, develop color at room temperature for 20 min in the da...

Embodiment 3

[0050] The purity identification of embodiment 3 egg yolk antibody

[0051] Get the product obtained in step (4) step (5) in the embodiment 1 and carry out yolk antibody purity identification, use non-reducing SDS-PAGE gel electrophoresis detection, the result is as follows figure 2 , M: Protein Maker; 1: crude extract of egg yolk antibody; 2: solution obtained after ammonium sulfate salting out; 3: solution obtained after sodium sulfate salting out; 4: solution obtained after ultrafiltration and concentration; 5: egg yolk antibody freeze-dried powder . Detected by reducing SDS-PAGE gel electrophoresis, the results are as follows image 3 , where M: protein maker; 1: crude extract of yolk antibody; 2: solution obtained after ammonium sulfate salting out; 3: solution obtained after sodium sulfate salting out; 4: solution obtained after ultrafiltration and concentration; 5: lyophilized egg yolk antibody pink. figure 2 The results show that the molecular weight of IgY is 180...

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Abstract

The invention discloses a compound preparation with egg yolk antibodies for resisting dairy cow mastitis and a method for preparing the compound preparation. The compound preparation comprises, by weight, 5-10 parts of egg yolk antibodies for resisting staphylococcus aureus, 5-10 parts of egg yolk antibodies for resisting Escherichia coli, 5-10 parts of egg yolk antibodies for resisting streptococcus agalactiae, 1-5 parts of food-grade pectin, 1-10 parts of multi-vitamins and 45-65 parts of deionized water. The compound preparation and the method have the advantages that the compound preparation is high in specificity and purity, excellent effects of inhibiting the staphylococcus aureus, the streptococcus agalactiae and the Escherichia coli can be realized, the antibodies are low in preparation cost and are convenient and simple, the compound preparation and the method are applicable to large-scale production, and the compound preparation can be used for preventing and treating the dairy cow mastitis due to the staphylococcus aureus, the streptococcus agalactiae and the Escherichia coli; the compound preparation is a paste-shaped preparation, paste can be directly injected into affected cow milk ponds and can be preserved in the affected cow milk ponds for a long term, and accordingly effects of sustained-release therapy can be realized.

Description

technical field [0001] The invention relates to the technical and application fields of biological products, in particular to an anti-cow mastitis egg yolk antibody compound preparation and a preparation method thereof. Background technique [0002] Cow mastitis, also known as cow mastitis, is a type of disease that causes an inflammatory response to the mammary glands of cows after being stimulated by physical, chemical, and microorganisms. diseases with the greatest economic losses. The main impact of mastitis in dairy cows is: the decline in milk production of dairy cows, the changed milk composition will also affect the quality of milk products, the high cost of waste milk, treatment of dairy cows mastitis and the huge economy brought by the elimination of sick cows loss. In 1996, according to the report of the American Cow Mastitis Association (NMC), about 35% of dairy cows in the world suffer from mastitis, and the milk production of sick cows decreases by about 20%,...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K39/40A61P31/04A61P15/14C07K16/12C07K16/02C07K1/34C07K1/30C07K1/14
Inventor 徐永平李晓宇耿慧君王丽丽甄宇红陈红莉吴妍妍
Owner DALIAN UNIV OF TECH
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