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A kind of large-scale production method of mycoplasma hyopneumoniae live vaccine

A technology of Mycoplasma hyopneumoniae and a production method is applied in the field of large-scale production of live Mycoplasma hyopneumoniae vaccine, and can solve problems such as no large-scale overall production method and the like

Active Publication Date: 2021-02-26
NANJING DAYAO NETWORK TECH CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

An invention applied by Nanjing Tianbang Biotechnology Co., Ltd.: a method for rapid freeze-drying of 50 heads of live vaccines for Mycoplasma porcine pneumonia, application number 201711096020.3, only reported the freeze-drying method for 50 heads, but no 50 heads or more than 100 heads Report on the overall production method of large scale

Method used

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  • A kind of large-scale production method of mycoplasma hyopneumoniae live vaccine
  • A kind of large-scale production method of mycoplasma hyopneumoniae live vaccine
  • A kind of large-scale production method of mycoplasma hyopneumoniae live vaccine

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0016] Embodiment 1 Mycoplasma pneumonia live vaccine (168 strains) antigen preparation

[0017] 1. Materials

[0018] 1.1 Strains: Live vaccine against Mycoplasma swine pneumonia (168 strains) was provided by Guizhou Foster Biotechnology Co., Ltd.

[0019] 1.2 Material: KM 2 The culture medium and related experimental consumables were provided by Guizhou Foster Biotechnology Co., Ltd., and the bacterial fermentation tank production workshop was provided by the School of Life Sciences, Hubei University.

[0020] 2. Method

[0021] Swine Mycoplasma pneumoniae live vaccine (168 strains) was subjected to step-by-step amplified fermentation culture according to the culture method of first-level seed liquid, second-level seed liquid, third-level seed liquid, 30L seed tank, and 300L culture tank, and finally set the culture tank The optimal design of the culture conditions was carried out sequentially according to a single variable, and a total of parameters were optimized, and t...

Embodiment 2

[0065] The semi-finished product test that embodiment 2 swine mycoplasma pneumonia live vaccine (168 strains) prepares

[0066] According to the above-mentioned optimum scheme conclusion of embodiment 1, test obtains that the culture titer of the live vaccine of mycoplasma swine pneumonia (168 strains) semi-finished product reaches 10 9 Every milliliter has passed the pure inspection and reached the standard of the current version of "Chinese Veterinary Pharmacopoeia". The specific requirements are as follows:

[0067] 1 Purity test is carried out according to the current version of "Chinese Veterinary Pharmacopoeia", and it should be pure.

[0068] 2 Viable bacteria counts take semi-finished products, in KM 2 The medium was serially diluted 10 times to 10 -12 , another set KM 2 Control 3 tubes of culture medium, make 3 replicates in the same batch, culture at 37°C for 10 days, check the discoloration of the culture medium, and check for the presence of typical Mycoplasma ...

Embodiment 3

[0069] Example 3 Determination of Large-scale Production Parameters of Porcine Mycoplasma Pneumonia Live Vaccine (168 Strains)

[0070] According to the protective agent formula concentration is 1.5wt.%, 3.0wt.%, 6.0wt.%, 9.0wt.%, 10.0wt.%, respectively, sucrose concentration is 12.5wt.%, 25wt.%, prepare the protective agent, At the same time, set the mass ratio of antigen and protective agent to 5:5, 6:4, 7:3, 8:2, and 9:1 respectively, and set the filling volume of 10mL vials to 2, 3, 4mL. Orthogonal experiments were performed to determine the bacterial counts of CCU prepared by different protocols, as shown in Table 6.

[0071] Set the freeze-drying program according to the following procedure, pre-freezing at -40°C for 4 hours, sublimation in the first stage at -40°C to -15°C for 28 hours, drying at -15°C to 28°C for 17 hours; or, pre-freezing at -40°C for 4 hours, First sublimation -40℃~-15℃, 18h, drying -15℃~28℃, 14h; -40℃ pre-freezing for 4h, first sublimation -40℃~-15...

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Abstract

A large-scale production method of Mycoplasma hyopneumoniae live vaccine, which is characterized in that it comprises the preparation of Mycoplasma hyopneumoniae antigen, the preparation of freeze-dried protective agent, the proportioning and mixing of antigen and protective agent, the freeze-dried composition of large-scale vaccine, and the final preparation Mycoplasma hyopneumoniae live vaccine. This large-scale production method can hold 50 or 100 large-scale volumes in each vial (7mL or 10mL in volume), and at the same time optimize the antigen titer of each head to reach ≥10 8 CCU (more than 100 times the national standard), has the advantages of good safety, simple preparation method, and reduced cost.

Description

technical field [0001] The invention relates to the technical field of veterinary biological products, and relates to a large-scale production method of a Mycoplasma hyopneumoniae live vaccine. Background technique [0002] Mycoplasma hyopneumonia, also known as swine asthma, is a contact chronic respiratory infectious disease caused by Mycoplasma hyopneumoniae, which exists widely in all parts of the world. Infection of pigs not only leads to lower feed conversion rate, poor growth and development, but also is prone to secondary virus or bacterial infection, resulting in increased mortality of pigs and heavy economic losses. Vaccination is the most important means of preventing this disease. [0003] In recent years, the application of live vaccines against Mycoplasma swine pneumonia has gradually been paid attention to and widely used. The traditional production of 10-head and 20-head products has frequent dilution steps in the process of immunization. An invention filed...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): A61K9/19A61K47/42A61K47/26A61K39/02A61P11/00A61P31/04C12N1/20C12R1/35
CPCA61K9/19A61K39/02A61K47/26A61K47/42A61K2039/552A61P11/00A61P31/04C12N1/20
Inventor 朱燕锋卢业云杨德明
Owner NANJING DAYAO NETWORK TECH CO LTD