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A composition for improving the stability of phage preparations and its application

A composition and phage technology, applied in the field of biotechnology and microbial products, can solve the problems of low survival rate, poor phage storage effect, and short phage storage time, and achieve the effects of cost reduction, good and stable protection, and high activity

Active Publication Date: 2022-05-27
QINGDAO PHAGEPHARM BIO TECH CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the existing protective agents have poor preservation effects on phages, the storage time of phages is short, the survival rate is low, and phages are easily contaminated, and must be preserved under low temperature and ultra-low temperature conditions, which greatly restricts the development of phages. Therefore, the storage technology of phage is an important technical problem restricting the wide application of phage

Method used

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  • A composition for improving the stability of phage preparations and its application
  • A composition for improving the stability of phage preparations and its application
  • A composition for improving the stability of phage preparations and its application

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0038] Example 1 Screening of Components of Compositions of the Invention

[0039] Select 12 kinds of common protective agent added components to carry out single factor experiments respectively: prepare and sterilize according to the concentration in Table 1, and set up a control group without adding protective agent, a total of 13 groups; among which groups 1-12 each protective agent The percentages are all g / L, and the balance is deionized water.

[0040] Table 1 Component concentrations of 13 phage protection agents and their sterilization methods

[0041]

[0042]

[0043] The protective agent components in the above Table 1 are uniformly mixed with Escherichia coli phage in a volume ratio of 1:1 under sterile conditions to obtain an Escherichia coli phage preparation. And placed at 37 ℃ for accelerated storage test at a constant temperature, the titer was detected, and the storage effect of each component was compared.

[0044] The results are shown in Table 2. U...

Embodiment 2

[0048] Example 2 Screening of the optimal component addition amount of the composition of the present invention

[0049] Among the above-mentioned protective agent components, the three kinds of magnesium sulfate, gelatin and potassium behenate with the best effect were screened for the working addition amount.

[0050] Magnesium sulfate

[0051] Under sterile conditions, different amounts of magnesium sulfate were mixed with Escherichia coli phage in a volume ratio of 1:1 to obtain an Escherichia coli phage preparation. And placed at 37 ℃ for accelerated storage test at constant temperature, the titer was detected, and the storage effects of different additions were compared.

[0052] The results are shown in Table 3. Under the storage temperature of 37 °C, the titers of 15g / L and 40g / L were reduced by 1 order of magnitude after the 5th day of storage, and 5 orders of magnitude after the 30th day of storage; it can be seen from Table 3 When the addition amount is between 20...

Embodiment 3

[0065] Example 3 Screening of the best combination of the composition of the present invention

[0066] The best protective agent components selected above are combined to screen the best composition.

[0067] Composition 1

[0068] Sodium chloride 6g / L, magnesium sulfate 30g / L, gelatin 5mL / L, glycine 0.1g / L, and water are prepared into composition 1 according to the above-mentioned component additions;

[0069] Composition 2

[0070] Add sodium chloride 6g / L, magnesium sulfate 30g / L, gelatin 5mL / L, glycine 0.1g / L, sodium dihydrogen phosphate 0.1g / L, dipotassium hydrogen phosphate 0.1g / L, and water according to the above components. Formulated into composition 2;

[0071] Composition 3

[0072] Sodium chloride 6g / L, magnesium sulfate 30g / L, gelatin 5mL / L, glycine 0.1g / L, sodium dihydrogen phosphate 0.1g / L, dipotassium hydrogen phosphate 0.1g / L, glycerin 20mL / L, water according to The above-mentioned component addition amount is formulated into composition 3

[0073] Comp...

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Abstract

The present invention relates to a composition for improving the stability of bacteriophage preparations and its application. Specifically disclosed is a composition for improving the stability of a phage preparation, comprising the following components: sodium chloride 6g / L, magnesium sulfate 20-35g / L, dipotassium hydrogen phosphate 0.1g / L, sodium dihydrogen phosphate 0.1g / L, Gelatin 3‑8mL / L, potassium sorbate 0.05‑0.2g / L, glycine 0.1g / L, glycerin 20mL / L, water balance. Also disclosed is the use of the composition for the preparation of phage formulations. Compared with other protective agent formulations, the composition formula of the present invention can keep the phage preparation product active for a long time and have a higher titer, and can maintain the stability of the phage preparation product for a longer time in a heat-accelerated environment and at room temperature. , greatly reducing the cost of transportation and warehousing.

Description

technical field [0001] The invention belongs to the field of biotechnology and microbial products, and particularly relates to a composition for improving the stability of a phage preparation and its application. Background technique [0002] The problems of drug resistance, ecological environment, drug residues in food, and bio-sanitation safety caused by the abuse of antibiotics have attracted worldwide attention. The Chinese government has also promulgated corresponding laws, regulations or measures to promote the "anti-resistance process". The research and development of new antibiotic substitutes has become a major scientific problem to be solved urgently in animal husbandry. As a naturally occurring bacterial "nemesis", bacteriophage may play an important role in the era of "resistance-free". Phages were used to treat human infections as early as 1917, but with the advent of antibiotics in the 1940s, research into their therapeutic value was largely put on hold. Bact...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N1/04C12R1/92
CPCC12N1/04
Inventor 潘强任慧英孙虎芝庄莹王佳存庄盈婷李大任张召佐
Owner QINGDAO PHAGEPHARM BIO TECH CO LTD
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