Vaccine composition and its preparation method and application

A vaccine composition and antigen technology, which is applied in the directions of drug combination, pharmaceutical formula, medical preparation containing active ingredients, etc., can solve the problems of not removing the immunosuppressive components of Mycoplasma hyopneumoniae, not using it, increasing the pollution of process steps, etc.

Active Publication Date: 2019-12-20
PU LIKE BIO ENG
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] Chinese patent application CN88101554A discloses that the membrane protein of Mycoplasma hyopneumoniae contains the antigenic protein of immunosuppressive components, and discloses a method of treating the membrane of Mycoplasma hyopneumoniae with a non-ionic surfactant, and then proceeds to obtain the non-immunosuppressive antigen. The technical solution, but the disclosed technical solution needs to obtain the Mycoplasma hyopneumoniae membrane first, and the process of obtaining the membrane requires freeze-thaw cycles, ultrasonic treatment, etc., and the operation process is cumbersome, and the antigen does not use the supernatant culture solution that does not contain Mycoplasma hyopneumoniae cells soluble antigen in
However, it does not remove the immunosuppressive components of Mycoplasma hyopneumoniae, and the treatment with protein A column increases the process steps and the possibility of contamination

Method used

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  • Vaccine composition and its preparation method and application
  • Vaccine composition and its preparation method and application
  • Vaccine composition and its preparation method and application

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0066] Example 1 Preparation of Mycoplasma hyopneumoniae

[0067] 1.1. Preparation of whole bacterial antigen of Mycoplasma hyopneumoniae

[0068] After unsealing the freeze-dried strain Mycoplasma hyopneumoniae HN0613, inoculate the liquid culture medium with 10% of the inoculum amount, shake culture at 37°C for 3 to 7 days, and harvest when the pH value drops from 7.5 to 6.8. seed. Take the first-grade seeds and inoculate the liquid medium with 5% inoculum amount, shake and culture at 37°C for 3-7 days, harvest when the pH value drops from 7.5 to 6.8, and use them as secondary production seeds after pure inspection. Inoculate the secondary seeds of qualified Mycoplasma hyopneumoniae HN0603 strain in liquid culture medium at 5% (v / v) respectively, shake and culture at 37°C for 3-7 days, and harvest when the pH value drops from 7.5 to 6.8. Make the final concentration of formaldehyde solution 0.3% (V / V), inactivate at 37°C for 24 hours, stir once every 4 hours, 10min each ti...

Embodiment 2

[0078] The preparation of embodiment 2 swine mycoplasma pneumonia vaccine composition

[0079] The mycoplasma hyopneumoniae antigen prepared by embodiment 1 and Montanide TM Gel 01 adjuvant is mixed together by the components and ratio contained in the mycoplasma pneumoniae vaccine composition in Table 3, stirred at a speed of 500-800r / min for 10-15min, and added 1% (volume ratio) thimerosal solution before terminating the stirring , so that the final concentration does not exceed 1 / 10,000, fully shake and mix, and store at 2-8°C after aliquoting.

[0080] Table 1 Preparation of Mycoplasma hyopneumoniae vaccine composition

[0081]

Embodiment 3

[0082] Example 3 Efficacy Test of Vaccine Compositions of Mycoplasma Pneumoniae Porcine with Different Components

[0083] The vaccine composition prepared in Example 2 was used to immunize 50 piglets at the age of 14-21 days respectively (excluding porcine reproductive and respiratory syndrome, porcine circular type 2 and swine fever), totally 12 groups, 5 piglets / group, 1mL / head. 42 days after immunization, inject 100 MID of virulent CVCC354 strain of Mycoplasma hyopneumoniae Jinan strain into all pig trachea 50 / head, observed 28 days after dissection, and observed lung lesions, lung lesions were scored according to the 28-point method. About 10 days after the challenge, pigs in the blank control group (3 of them) successively developed symptoms such as coughing and wheezing, and their fur was not smooth;

[0084] Table 2 The scores of lung lesions and the reduction rate of lung lesions in each test group

[0085] Group No immune components Mean pneumonia l...

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Abstract

The present invention provides a method for preparing a vaccine composition containing Mycoplasma hyopneumoniae antigen, wherein the method comprises: (1) culturing and inactivating Mycoplasma hyopneumoniae; (2) treating step (1) with a nonionic surfactant inactivated Mycoplasma hyopneumoniae; and (3) separating the non-ionic surfactant-insoluble antigen components after treatment with the non-ionic surfactant, and adding an adjuvant. The present invention also provides that the vaccine composition containing Mycoplasma hyopneumoniae antigen prepared by the preparation method removes the immunosuppressive components, and at the same time retains the immune active components to the greatest extent; It has a good protective effect on the lung disease of pigs.

Description

technical field [0001] The invention belongs to the technical field of veterinary biological products, and in particular relates to a vaccine composition containing swine mycoplasma pneumonia antigen and a preparation method thereof. Background technique [0002] Mycoplasma pneumonia of swine (Mycoplasmalpneumonia of swine, MPS) is also known as swine panting disease (Li Yanming, Zhang Ying. Advances in biological research on Mycoplasma hyopneumoniae, Advances in Veterinary Medicine, 2003,24(3):25-27) or swine endemic Pneumonia is a contact chronic respiratory disease of pigs caused by Mycoplasma hyopneumoniae (Mhp). It is widely distributed and characterized by chronicity, contact, high infectivity, high incidence and low mortality. Manifested as cough and dyspnea, dissection showed fleshy or marbled lung tissue lesions. [0003] The Ross study at the University of Iowa found that after Mhp infection, the ability of lymphocytes to produce antibodies decreased, cellular imm...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): A61K39/02A61K39/295A61K39/108A61P31/04A61P11/00A61P31/20
Inventor 张许科孙进忠廖永洪田克恭
Owner PU LIKE BIO ENG
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