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A kind of method utilizing serum-free culture medium to produce pedv attenuated vaccine

A technology of serum-free culture medium and attenuated vaccine is applied in the field of PEDV attenuated vaccine production, which can solve the problems of high cost of serum production, and achieve the effect of avoiding immune problems and promoting proliferation.

Active Publication Date: 2019-07-02
陕西诺威利华生物科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0007] In order to solve the above technical problems, as well as the many problems existing in the use of serum in the prior art and the high production cost, the present invention aims to provide a method for producing attenuated PEDV vaccine using a serum-free medium without adding animal serum, It greatly reduces the production cost, improves the economic benefits of production, and avoids many problems caused by the use of serum

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  • A kind of method utilizing serum-free culture medium to produce pedv attenuated vaccine
  • A kind of method utilizing serum-free culture medium to produce pedv attenuated vaccine

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Effect test

Embodiment 1

[0044] Embodiment 1: a kind of serum-free medium suitable for large-scale production of PEDV vaccine is characterized in that, described serum-free medium is made of basal medium, bovine serum albumin, cortisol, insulin, transferrin, glutathione It consists of peptide, trypsin, taurine and sodium selenite, wherein the weight ratio of taurine and sodium selenite is 2:1.

[0045] The basal medium IMDM (Giboc);

[0046] The content of the bovine serum albumin is 10mg / mL.

[0047] The content of the cortisol is 20 μg / mL.

[0048] The content of the insulin is 20 μg / mL.

[0049] The content of the transferrin is 15 μg / mL.

[0050] The content of glutathione is 3mM.

[0051] The content of the trypsin is 4 μg / mL.

[0052] The content of the taurine is 20 μg / mL.

[0053] The content of the sodium selenite is 10 μg / mL.

[0054]The preparation method of the serum-free medium that is suitable for large-scale production of PEDV vaccine of the present invention is: step 1, take by ...

Embodiment 2

[0055] Embodiment 2: a kind of serum-free medium suitable for large-scale production of PEDV vaccine is characterized in that, described serum-free medium is made of basal medium, bovine serum albumin, cortisol, insulin, transferrin, glutathione It consists of peptide, trypsin, taurine and sodium selenite, wherein the weight ratio of taurine and sodium selenite is 3:1.

[0056] The basal medium IMDM (Giboc);

[0057] The content of the bovine serum albumin is 10mg / mL.

[0058] The content of the cortisol is 20 μg / mL.

[0059] The content of the insulin is 20 μg / mL.

[0060] The content of the transferrin is 15 μg / mL.

[0061] The content of glutathione is 3mM.

[0062] The content of the trypsin is 4 μg / mL.

[0063] The content of the taurine is 30 μg / mL.

[0064] The content of the sodium selenite is 10 μg / mL.

[0065] The preparation method of the serum-free medium that is suitable for large-scale production of PEDV vaccine of the present invention is: step 1, take by...

Embodiment 3

[0066] Embodiment 3: PEDV small-scale proliferation test

[0067] Test strain: PEDV SX-YL strain, obtained from Northwest Agriculture and Forestry University, which was isolated from a large-scale farm in Yangling, Shaanxi.

[0068] Experimental cells: Vero cell line, a gift from the Veterinary Immunology Laboratory, School of Veterinary Medicine, Northwest A&F University.

[0069] experiment method:

[0070] (1) Recovery of Vero cell lines:

[0071] (1) Quickly put Vero cells taken out of liquid nitrogen into a 37°C water bath, shake gently to thaw quickly;

[0072] (2) Centrifuge at 300 rpm for 10 min;

[0073] (3) Discard the supernatant, add 37°C pre-warmed cell culture medium (containing 5% FBS DMEM), and gently blow off the Vero cells;

[0074] (4) Add the mixed cell suspension into a sterile T25 bottle containing 5m L5% FBS DMEM cell culture medium, and culture it in a cell culture incubator with suitable conditions;

[0075] (5) After two passages, add the vero ce...

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Abstract

The invention relates to a method for producing a PEDV attenuated vaccine by using a serum-free culture medium. The method is characterized in that the serum-free culture medium is prepared from a basal culture medium, bovine serum albumin, cortisol, insulin, transferrin, glutathione, trypsin, taurine and sodium selenite, wherein the proportioning ratio (in parts by weight) of the taurine to the sodium selenite is (2 to 3): 1. The culture medium is free of animal blood serum, so that the production cost is reduced greatly, the economic benefit of production is improved, and many problems resulting from blood serum employment are also avoided.

Description

[0001] Technical field: [0002] The invention relates to the field of biotechnology, in particular to a method for producing PEDV attenuated vaccine by using a serum-free medium. [0003] Background technique: [0004] Porcine epidemic diarrhea (PED) is an acute and highly contagious enteroviral disease caused by porcine epidemic diarrhea virus (PEDV), the symptoms of which include diarrhea, vomiting, anorexia, dehydration and weight loss in piglets. Although pigs of different ages may be infected to varying degrees and develop symptoms, the condition of piglets is particularly severe, and the mortality rate is as high as 100%. PEDV mainly enters the host through the intestinal mucosal surface of piglets, mainly causing damage to intestinal epithelial cells. Although pigs of different ages may be infected to varying degrees and develop symptoms, the condition of piglets is particularly severe, and the mortality rate is as high as 100%. PEDV mainly enters the host through the i...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): A61K39/215A61P31/14C12N7/00
CPCA61K39/12A61K2039/5254A61P31/14C12N7/00C12N2770/20034C12N2770/20051
Inventor 杜恩岐陈瑞
Owner 陕西诺威利华生物科技有限公司
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