Culture medium for screening strain capable of highly yielding Hib vaccine as well as preparation method and screening method of strain

A technology of culture medium and solid medium, which is applied in the biological field to achieve the effects of increased production, low cost and convenient preparation

Inactive Publication Date: 2015-01-21
CHENGDU OLYMVAX BIOPHARM
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the ease with which capsulated Haemophilus influenzae type b strains mutate to the non-capsulated form can pose a problem for PRP production, therefore, how to screen high-producing Hib vaccine strains has become the focus of research in this field

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0034] Embodiment 1: preparation culture medium

[0035] A solid medium for screening high-yield Hib vaccine strains, the formula includes the following components: relative to the amount of solvent used as 1000ml, it also contains 10g of yeast extract, 3g of disodium hydrogen phosphate, 0.94g of sodium dihydrogen phosphate, 60 % sodium lactate solution 0.5ml, magnesium chloride 0.2g, calcium chloride dihydrate 0.01g, ammonium sulfate 0.5g, glucose 10g, hemin 10mg, coenzyme I 10mg, agar 10g, adjust the pH to 6.5, and the solvent is water;

[0036] The preparation method of above-mentioned solid culture medium comprises the following steps:

[0037] S1. Weighing: Weigh the raw materials of each component according to the ratio of the above solid medium formula, and set aside;

[0038] S2. Preparation of liquid A:

[0039] S21. Ultrafiltration: Dilute the yeast extract with distilled water to a concentration of 100g / L, dilute the glucose with distilled water to a concentration...

Embodiment 2

[0043] Embodiment 2: preparation culture medium

[0044] A solid medium for screening high-yielding Hib vaccine strains, the formula includes the following components: relative to the amount of solvent used in 1000ml, it also contains 20g of yeast extract, 28.65g of disodium hydrogen phosphate, 3.12g of sodium dihydrogen phosphate, 1.5ml of 60% sodium lactate solution, 0.6g of magnesium chloride, 0.04g of calcium chloride dihydrate, 1g of ammonium sulfate, 20g of glucose, 20mg of hemin, 20mg of coenzyme I, 15g of agar, adjust the pH to 7.5, and the solvent is water;

[0045]The preparation method of above-mentioned solid culture medium comprises the following steps:

[0046] S1. Weighing: Weigh the raw materials of each component according to the ratio of the above solid medium formula, and set aside;

[0047] S2. Preparation of liquid A:

[0048] S21. Ultrafiltration: Dilute the yeast extract with distilled water to a concentration of 90g / L, dilute the glucose with distille...

Embodiment 3

[0052] Embodiment 3: preparation culture medium

[0053] A liquid medium for screening high-yield Hib vaccine strains, the formula includes the following components: yeast extract 14g, disodium hydrogen phosphate 10g, sodium dihydrogen phosphate 1.83g, 60% sodium lactate solution 0.8ml, magnesium chloride 0.3g, dihydrogen phosphate Calcium chloride 0.02g, ammonium sulfate 0.72g, glucose 14g, hemin 12mg, coenzyme I 13mg, adjust the pH to 7, and the solvent is water.

[0054] The preparation method of above-mentioned liquid culture medium comprises the following steps:

[0055] (1) Weighing: take the raw materials of each component according to the ratio of the above-mentioned liquid culture medium formula, and set aside;

[0056] (2) Preparation of liquid A:

[0057] A. Ultrafiltration: Dilute the yeast extract with distilled water to a concentration of 110g / L, dilute the glucose with distilled water to a concentration of 220g / L, perform ultrafiltration with a membrane bag wi...

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PUM

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Abstract

The invention discloses a culture medium for screening a strain capable of highly yielding a Hib vaccine. The formula of the culture medium comprises the following components: a yeast extract, disodium hydrogen phosphate, sodium dihydrogen phosphate, a 60% sodium lactate solution, magnesium chloride, calcium chloride dihydrate, ammonium sulfate, glucose, chlorhematin, a coenzyme I and/or agar. A preparation method of the culture medium for screening the strain capable of highly yielding the Hib vaccine comprises the steps of weighting, preparing a solution A, preparing a solution B and mixing. The high-yielding strain with a capsule is screened by virtue of carrying out passage growth on solid and liquid culture media for many times. Since the culture medium disclosed by the invention is free of animal blood and protein components derived from animals, the potential risk is avoided and the safety of the vaccine is improved; the preparation method of the culture medium has the advantages of simple operation, convenience in preparation and low cost; due to the screening method, the problem of strain screening in the scale production of the Hib vaccine is solved, the yield of the Hib capsular polysaccharide is greatly increased, the working efficiency is improved and the production cost is decreased.

Description

technical field [0001] The invention belongs to the field of biotechnology, and in particular relates to a culture medium for screening high-yield Hib vaccine strains, a preparation method and a screening method. Background technique [0002] Haemophilus influenzae type b (Hib) is a type of Haemophilus influenzae with extremely strong invasiveness. Among the serious infections caused by Haemophilus influenzae, almost 90% are caused by type B, mainly causing meningitis, Pneumonia, pericarditis, arthritis, bacteremia, epiglottitis and other diseases, the disease makes 15% to 35% of survivors suffer from permanent disability, typical symptoms include mental retardation or deafness. Meningitis and pneumonia caused by Haemophilus influenzae type b can lead to mental retardation or deafness. Meningitis and pneumonia caused by Haemophilus influenzae type b are the leading causes of infant mortality. Years of epidemiological studies have shown that the use of Hib vaccine is an eff...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N1/20C12R1/21
CPCC12N1/20
Inventor 陈道远朱冲陈元芬
Owner CHENGDU OLYMVAX BIOPHARM
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