Method for producing swine erysipelas live vaccine

A production method and technology for vaccine production, applied in bacterial antigen components, antibacterial drugs, etc., can solve the problems of low freeze-drying survival rate, instability between batches, storage, transportation and use difficulties, etc.

Active Publication Date: 2015-01-28
北京中海生物科技有限公司 +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the quality of meat, liver, and gastric extract, the main components of meat liver stomach (membrane) digestion soup, is greatly affected by the type, age, freshness, and degree of digestion of the animal. The number of cultured bacteria is low and unstable between batches.
(2) Vaccine freeze-drying adopts 1.5% gelatin 5% sucrose as protective agent all the time, and th

Method used

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  • Method for producing swine erysipelas live vaccine
  • Method for producing swine erysipelas live vaccine
  • Method for producing swine erysipelas live vaccine

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0066] - Preparation of synthetic media

[0067] 1. Preparation of Synthetic Medium

[0068] Synthetic medium formula (W / V) is as follows:

[0069] Soytone 10g, Peptone 20g, Yeast Powder 20g, Dipotassium Hydrogen Phosphate 0.675g, Potassium Dihydrogen Phosphate 0.1632g, Magnesium Sulfate 0.2g, Glucose 5g, L-Arg-HCl 5g, Tryptophan 1g, Tween-800.5ml 1. Add water for injection to 1000ml;

[0070] 2. Preparation method of synthetic medium

[0071] According to the above formula, the culture medium components (except glucose) were weighed sequentially and added to the water for injection. After fully dissolving, adjust the pH value to 7.8-8.2 with 2mol / L sodium hydroxide solution, and sterilize at 116°C for 30 minutes. Just before use, aseptically add 50% glucose solution with a final concentration of 0.5%, and mix well.

[0072] 50% glucose solution (W / V) Weigh 50g of glucose, add water for injection to dissolve, dilute to 100ml, sterilize at 116°C for 30 minutes, store at 2-8...

Embodiment 2

[0075] ——Preparation of lyoprotectant

[0076] The formulation of the protectant

[0077] Dextran 10g, BSA 1g, sorbitol 50g, sucrose 50g, hydrolyzed milk protein 5g, L-sodium glutamate 5g, dipotassium hydrogen phosphate 1.25g, potassium dihydrogen phosphate 0.52g, water for injection 1000ml.

[0078] 2. Preparation method

[0079] Solution 1: In a clean beaker, add an appropriate amount of water for injection (100°C), weigh dextran, BSA, and sorbitol in turn according to the formula, add them to the beaker, and stir with a glass rod while adding to make it completely dissolve, then use Dilute to 500ml with water for injection, and sterilize at 116°C for 30 minutes.

[0080] Solution 2: Weigh sucrose, hydrolyzed milk protein, sodium L-glutamate, potassium dihydrogen phosphate and dipotassium hydrogen phosphate in a beaker in turn according to the formula, add water for injection to fully dissolve it, and set the volume to 500ml, 0.22μm Membrane-sterilizing filters are steril...

Embodiment 3

[0083] ——Erysipelas live vaccine (CVCC1318)

[0084] 1 Preparation of bacterial solution for seedling production

[0085] Inoculate the Erysipelas suis CVCC1318 seed liquid into the liquid medium according to 1% to 2% of the total medium, ferment or aerate at 37°C for 8 to 10 hours, add an appropriate amount of antifoaming agent according to the need during the cultivation process, and When the pH rises, add an appropriate amount of sterilized 2mol / L sodium hydroxide solution to control the pH value. After the cultivation, the bacterial solution was centrifuged at 3500r / min for 20min, and the bacterial precipitate was harvested. The precipitate was suspended by adding an appropriate amount of heat-resistant freeze-drying protective agent that had been sterilized and preheated to 37°C. During the subpackaging process, attention should be paid to keeping warm and shaking evenly. After subpackaging, freeze and vacuum dry quickly, according to the method stipulated in the "Chines...

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Abstract

The invention relates to application of a synthetic medium and a freeze-drying protective agent in the production of a swine erysipelas live vaccine. The synthetic medium provided by the invention is designed according to growth characteristics and growth requirements of bacillus erysipelatos-suis, is controllable in raw material sources of the formula, is not influenced by materials such as beef, beef liver, pig stomach and so on, is stable in batch, simple in manufacturing, convenient to use, suitable for carrying out high cell density culture of bacteria and high in amount of culture bacteria; in the freeze-drying process, through the adoption of the freeze-drying protective agent developed for the characteristics of the bacillus erysipelatos-suis and a matched freeze-drying curve, the activity of the bacteria is effectively protected in the freeze-drying process, so that the survive rate of the freeze-dried bacteria is increased and the vaccine is preserved for a long time at 2-8 DEG C and can be preserved for 24 months.

Description

[0001] Technical field The present invention relates to a production method of porcine erysipelas live vaccine, which belongs to the field of biology, especially the field of veterinary biological products. Background technique [0002] Erysipelas is one of the three major infectious diseases of pigs. It is distributed worldwide and is also a zoonotic disease. The pathogen is Erysipelas suis. At present, the existing swine erysipelas live vaccines in our country include single vaccine, double vaccine and triple vaccine, and they have played an important role in preventing swine erysipelas disease. But at present, there are the following outstanding problems in the bacterial culture and freeze-drying process, which is far behind developed countries. (1) Poor culture stability. From the 1960s to the 1970s, the meat liver and stomach (membrane) digestion soup containing 2% to 4% lysed whole blood was used as the medium for the production of vaccines. Since the 1980s, domestic ve...

Claims

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Application Information

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IPC IPC(8): A61K39/02A61P31/04
Inventor 朱良全李聪研丁家波孙晔沈青春
Owner 北京中海生物科技有限公司
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