Triple yolk antibody freeze-dried powder as well as preparation method and application thereof

A technology of egg yolk antibody and freeze-dried powder, which is applied in the direction of antibody, freeze-dried transportation, and veterinary vaccines. low effect

Active Publication Date: 2021-08-13
SOUTHWEST UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The low production cost, easy preparation, strong pertinence, and good curative effect of yolk antibody make it widely welcomed by farm technicians. The promotion and application of various yolk antibody products

Method used

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  • Triple yolk antibody freeze-dried powder as well as preparation method and application thereof
  • Triple yolk antibody freeze-dried powder as well as preparation method and application thereof
  • Triple yolk antibody freeze-dried powder as well as preparation method and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0015] Preparation method of anti-pig PRV-TGEV-PEDV triple yolk antibody freeze-dried powder

[0016] 1) Use 3mL TGEV, 2mL PRV and 3mL attenuated strain antigens of PEDV (each antigen diluted 1:8 with aluminum hydroxide adjuvant and then stored at 37°C for 20 minutes), subcutaneously immunize the neck at 140 days in sequence For laying hens, each immunization interval is 7 days, each antigen is immunized 4 times, and the total immunization time is 3 months. The immunized eggs are collected, egg yolk and egg white are separated, and PBS solution (0.01 mL / L, pH7.2), add PEG-6000 so that the final concentration of PEG-6000 in the solution is 3.5%, stir evenly, place at 4°C for 20min, and centrifuge at 5000rpm for 20min. Take the supernatant and add PEG-6000, so that the concentration of PEG-6000 in the solution is 12%, mix well and then let it stand for 20 minutes, centrifuge at 5000rpm for 20 minutes, pour off the supernatant, leave the precipitate and dissolve it in PBS Store ...

Embodiment 2

[0019] Design and Screening of Lyoprotectant Components

[0020] (1) Design of lyoprotectant components

[0021] Three kinds of lyoprotectants were prepared from gelatin, sucrose and other ingredients, respectively numbered A-1, A-2, A-3, and prepared triple yolk antibody at the ratio of 1:1, 1:1.5, and 1:2 respectively. The solution was evenly mixed, and 0.01% thimerosal was added as a preservative. 3 kinds of protective agents with different components were mixed with the triple egg yolk antibody solution in 3 proportions to form 9 groups of test samples. Each group set 3 parallel samples and numbered them. A represents the formula of the freeze-dried protective agent, and B represents different Adding ratio, B1 represents 1:1; B2 represents 1:1.5, B3 represents 1:2. See Table 1 for specific groups.

[0022] Table 1 Lyoprotectant components (unit: %)

[0023]

[0024] (2) Screening of optimal lyoprotectant formulations

[0025] ① Screening criteria

[0026] The qual...

Embodiment 3

[0040] Setting and screening of freeze-dried powder storage conditions

[0041] (1) Setting of freeze-dried powder temperature

[0042] Place the vials in a refrigerator at -20°C for 4 hours, and at -80°C for 12 hours, then put them in a freeze-vacuum dryer (-54°C) for 30 hours to make a freeze-dried preparation. The vacuum degree is maintained at 10-30Pa. Keep at -80°C. Store in a -20°C refrigerator.

[0043] (2) Screening of storage conditions for freeze-dried powder of anti-porcine PRV-TGEV-PEDV triple yolk antibody

[0044] Take the freeze-dried powder of the formula with the best antibody titer, set three storage temperature values, namely 4°C, 15-25°C (room temperature environment), and -20°C, set two parallel samples for each temperature value, and store them for 6 When the storage time expires, the samples stored at -20°C shall be thawed naturally at 4°C, and the samples stored at 4°C and 15°C shall be referred to the antibody titer detection methods and standards i...

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Abstract

The invention relates to triple yolk antibody freeze-dried powder as well as a preparation method and application thereof, and belongs to the technical field of veterinary drugs. The triple yolk antibody freeze-dried powder is stored for 6 months at the temperature of -20 DEG C, and the antibody titer is basically unchanged. The preparation method is low in cost and simple in operation process, fat and other impurities in yolk liquid are easier to remove compared with other methods, and the influence on the yolk antibody titer in the yolk liquid is minimum.

Description

technical field [0001] The invention belongs to the technical field of veterinary medicine, and in particular relates to a triple yolk antibody freeze-dried powder, a preparation method and application thereof. Background technique [0002] The rapid development of large-scale pig farming in China is accompanied by more and more complex diseases. In recent years, porcine transmissible gastro-enteritis virus (TGEV), porcine epidemic diarrhea virus (PEDV) and pseudorabies virus (PRV) have The main pathogen of intractable diarrhea in piglets in large-scale pig farms has caused huge economic losses to farmers. Vaccination is the main measure to prevent and control the three diseases. [0003] Yolk antibody solution (IgY) is an immunoglobulin with no residue and no side effects, and is widely used in the prevention and treatment of diseases. There are many reports of piglet diarrhea secondary to immune failure caused by various reasons in large-scale pig farms, which provides ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K39/42A61K9/19A61K47/42A61K47/26A61K47/36A61K47/32A61P31/14
CPCA61K39/42A61K9/19A61K47/42A61K47/26A61K47/36A61K47/32A61P31/14A61K2039/70A61K2039/552Y02A50/30
Inventor 魏光河孟霞李海帆姚馨月杨萍瑞
Owner SOUTHWEST UNIVERSITY
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