CN105950570A - Preservation method for bacteriophage - Google Patents
Preservation method for bacteriophage Download PDFInfo
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- CN105950570A CN105950570A CN201610422049.5A CN201610422049A CN105950570A CN 105950570 A CN105950570 A CN 105950570A CN 201610422049 A CN201610422049 A CN 201610422049A CN 105950570 A CN105950570 A CN 105950570A
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N7/00—Viruses; Bacteriophages; Compositions thereof; Preparation or purification thereof
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N2795/00—Bacteriophages
- C12N2795/00011—Details
Abstract
The invention provides a preservation method for bacteriophage. The preservation method comprises the steps that bacteriophage liquid and a bacteriophage preservation protecting agent are mixed to be uniform to obtain a bacteriophage stock solution, and then the bacteriophage stock solution is subpackaged into ampoules to be sealed and cryo-preserved at room temperature or the temperature of 2 DEG C to 7 DEG C. The preservation method for the bacteriophage is reasonable and convenient to apply, and the bacteriophage can be preserved at the room temperature or refrigerated, is low in requirement for the preservation environment and good in adaptability and can be preserved for a long term.
Description
Technical field
The invention belongs to biotechnology and microbial product field, in particular it relates to a kind of phagocytosis
The store method of body.
Background technology
Phage is the total of the virus of the microorganisms such as a class energy bacterial infection, actinomycetes, spirillum
Claim, some antibacterials are had the specificity of height, when these antibacterials of Phage Infection, can be
Antibacterial breeds and kills antibacterial, and it does not has toxicity to animals and plants.Along with molecular biology skill
The development of art, phage is widely used for multiple fields such as medical diagnosis on disease, water process.Due to
Gene can be inserted the characteristic in host DNA by phage so that it also becomes important
Molecule and genetics research instrument.Utilize phage, a lot of exquisite experiment can be designed.
But in above process, be required for phage is preserved, the preservation of prior art
The preservation effect of scheme pnagus medius is poor, and the holding time of phage is short, and survival rate is low, phagocytosis
Body is easily contaminated, and must preserve under cryogenic, seriously constrains phagocytosis
The development of body application.
Summary of the invention
Goal of the invention: in order to overcome above deficiency, the invention provides the preservation of a kind of phage
Method.
Technical scheme: the invention provides the store method of a kind of phage, comprise the following steps:
1) phage solution stands 4-8h in 2-7 DEG C, flocculent deposit occurs, passes through
8000-12000r/min is centrifuged 20-40min；
2) in above-mentioned precipitation, add phage buffer, by resolution of precipitate, obtain phagocytosis body fluid；
3) phage of phagocytosis body fluid volume 1-2.5 times preserves protective agent mix homogeneously and obtains phagocytosis
Body stock solution, described phage preserves aminoacid described in protective agent by weight part, including following
Component: defatted milk powder 5-12 part, peptone 3-5 part, sucrose 0.5-1.2 part, trehalose
0.8-1.5 part, lentinan 1-1.6 part, magnesium sulfate 0.2-0.5 part, calcium chloride 0.6-1.2
Part, sodium sulfate 0.6-0.8 part, gelatin 0.05-0.2 part, mannitol 0.8-1.2 part, poly-second
Alkene pyrrolidone 0.1-0.3 part, starch 0.5-0.8 part, Pulvis Talci 0.3-0.5 part, carapace
Element 0.08-0.2 part, potassium dihydrogen phosphate 0.03-0.3 part, disodium hydrogen phosphate 0.05-0.2 part,
Glycerol 1-3 part, tween 80 0.02-0.1 part and 100 parts of water；
4) above-mentioned phage stock solution is sub-packed in ampoule sealing, enters in room temperature or 2-7 DEG C
Row is frozen.
The store method of phage of the present invention, method is reasonable, and application is convenient, phage
Stock solution adds and preserves protective agent, can be stored at room temperature or cold preservation, to Conservation environment requirement
Low, adaptability is good, can preserve for a long time.It is reasonable that its pnagus medius preserves protectant component,
Wherein defatted milk powder, peptone, gelatin, polyvinylpyrrolidone and starch etc. form whole guarantor
Deposit protectant skeleton, prevent carbonization and the oxidation of lower-molecular substance, protect active substance simultaneously
Not temperature influence, makes protective agent form porous, loose sponge and have certain
Water content, keeps original framework during frozen, thus increases dissolubility.Additionally, it is many
Plant saccharide and salt phage keeps existing state and moisture rises mitigation, and keep electricity
Solve matter concentration.Phage preserves protective agent can make phage damage from solution damage and ice crystal
Etc. the damage of physical factor, and maintain the high survival rate of phage and activity, make phage
Activity be in half static or resting state.Additionally, various saccharides and salt make phage keep depositing
Live state and moisture is risen mitigation, and keep electrolyte concentration.Phage preserves
Protect agent can make under phage cryopreservation from physical factors such as solution damage and ice crystal damages
Damage, and maintain the high survival rate of phage and activity, make the activity of phage be in
Half static or resting state.Additionally, protective agent is with the water molecules in solution, it is hydrated
Effect, the crystallization process of reduction water makes the viscosity of solution increase thus reduces the formation of ice crystal, with
Time cryoprotective agent can reduce inside and outside by maintaining certain molar concentration inside and outside phage
Do not freeze the concentration of electrolyte in solution, makes cell from the damage of solute.Additionally, protective agent
Can also be used for the preservation of multiple phage at ambient temperature, adaptability is good.
Further, the store method of above-mentioned phage, described phage preserve protective agent with
Components by weight percent meter, also includes the aminoacid of 0.3-0.8 part, described aminoacid by weight part,
Including following components: polyglutamic acid 1-4 part, aspartic acid 3-5 part, phenylalanine 2-8
Part, serine 2-5 part, cysteine 0.5-2 part, methionine 3-5 part, lysine 2-4
Part, leucine 0.3-0.8 part and isoleucine 0.2-0.6 part.By the liquid that has additional nutrients, make
Phage keeps existing state and moisture is risen mitigation.
Further, the store method of above-mentioned phage, described ampoule is put into equipped with anhydrous chlorine
Change in the sealing container of calcium or absolute methanol or silica gel in room temperature or 2-7 DEG C of preservation.Dry
Under the conditions of preservation effect more preferably, the holding time is more long.
Further, the store method of above-mentioned phage, described step is in the humidity of 4-6%
Under the conditions of complete.Under the damp condition of 4-6%, Bacteriolytic activity is optimal, and preservation effect is best.
Further, the store method of above-mentioned phage, described phage buffer is with weight
Component meter, including following components: Tri(Hydroxymethyl) Amino Methane Hydrochloride 8-15 part, sodium chloride
0.05-0.2 part, potassium chloride 1-5 part, magnesium sulfate 5-20 part, dipotassium hydrogen phosphate 1-3 part,
Sodium dihydrogen phosphate 8-12 part, gelatin 0.05-0.1 part and 100 parts of water, and regulate pH
To 7-7.5.Phage buffer composition is reasonable, easy to use, may be used for the molten of phage
Solve and resuspended, and do not affect the activity of phage.
Further, the store method of above-mentioned phage, described starch be potato starch,
The one of tapioca, corn starch, green starch or hetastarch.The raw material of starch comes
Source is wide, and selectivity is good, and application cost is low.
Further, the store method of above-mentioned phage, described peptone is phytone.
In phytone, the content of cellulose in the cell wall of plant cell is many, can be formed preferably
Framing structure, and the raw material of plant origin to compare the raw material of animal origin cheap, application cost
Low.
Further, the store method of above-mentioned phage, described glycerol is 20%-40%'s
Glycerol.The glycerol good fluidity of 20%-40%, it is easy to application.
Further, the store method of above-mentioned phage, described water is deionized water or pure
Change water.Deionized water or purified water can ensure that phage preserve during steady quality, controlled.
Further, the preparation method of the store method of above-mentioned phage, use dilute hydrochloric acid and
/ or sodium hydrate regulator solution pH.Raw material sources are wide, regulate very convenient, and are applied to
This is low.
Beneficial effect: compared with prior art, the invention have the advantages that of the present invention
The store method of phage, method is reasonable, and application is convenient, and phage can be stored at room temperature
Or cold preservation, requires low to Conservation environment, and adaptability is good, and can preserve for a long time.
Accompanying drawing explanation
Fig. 1 is the phage survival rate that use embodiment 1-5 store method of the present invention preserves
Figure.
Detailed description of the invention
To be further elucidated with the present invention by several specific embodiments below, these embodiments are only
Illustrate that problem, be not a kind of restriction.
Embodiment 1
The store method of a kind of coliphage, completes under the damp condition of 4%, bag
Include following steps:
1) coliphage solution stands 4h in 2 DEG C, flocculent deposit occurs, passes through
8000r/min is centrifuged 20min；
2) in above-mentioned precipitation, add phage buffer, by resolution of precipitate, obtain phagocytosis body fluid；
Described phage buffer by weight part, including following components: trishydroxymethylaminomethane
Hydrochlorate 8 parts, 0.05 part of sodium chloride, 1 part of potassium chloride, 5 parts of magnesium sulfate, phosphoric acid hydrogen
Dipotassium 1 part, sodium dihydrogen phosphate 8 parts, 0.05 part of gelatin and deionized water 100 parts, and
And use dilute hydrochloric acid and/or sodium hydroxide regulation pH to 7；
3) the isopyknic phage of phagocytosis body fluid preserves protective agent mix homogeneously and obtains phage stock solution,
Described phage preserves aminoacid described in protective agent by weight part, including following components: de-
Fat milk powder 5 parts, phytone 3 parts, sucrose 0.5 part, trehalose 0.8 part, Lentinus Edodes
Polysaccharide 1 part, 0.2 part of magnesium sulfate, 0.6 part of calcium chloride, 0.6 part of sodium sulfate, gelatin 0.05
Part, 0.8 part of mannitol, polyvinylpyrrolidone 0.1 part, potato starch 0.5 part,
Pulvis Talci 0.3 part, chitin 0.08 part, potassium dihydrogen phosphate 0.03 part, disodium hydrogen phosphate
0.05 part, the glycerol 1 part of 20%, tween 80 0.02 part and deionized water 100 parts；
Additionally, also include the aminoacid of 0.3 part, described aminoacid by weight part, including
Following components: polyglutamic acid 1 part, aspartic acid 3 parts, phenylalanine 2 part, thread ammonia
Acid 2 parts, cysteine 0.5 part, methionine 3 parts, lysine 2 parts, leucine 0.3
Part and isoleucine 0.2 part；
4) above-mentioned phage stock solution being sub-packed in ampoule sealing, described ampoule is put into equipped with nothing
In 15 DEG C of preservations in the sealing container of water calcium chloride.
Embodiment 2
The store method of a kind of mycobacterium phage, completes under the damp condition of 6%, bag
Include following steps:
1) mycobacterium phage solution stands 8h in 7 DEG C, flocculent deposit occurs, passes through
12000r/min is centrifuged 40min；
2) in above-mentioned precipitation, add phage buffer, by resolution of precipitate, obtain phagocytosis body fluid；
Described phage buffer by weight part, including following components: trishydroxymethylaminomethane
Hydrochlorate 15 parts, 0.2 part of sodium chloride, 5 parts of potassium chloride, 20 parts of magnesium sulfate, phosphoric acid hydrogen
Dipotassium 3 parts, sodium dihydrogen phosphate 12 parts, 0.1 part of gelatin and purified water 100 parts, and
Use dilute hydrochloric acid and/or sodium hydroxide regulation pH to 7.5.
3) phage of phagocytosis body fluid volume 2.5 times preserves protective agent mix homogeneously and obtains phage
Stock solution, described phage preserves aminoacid described in protective agent by weight part, including following group
Point: defatted milk powder 12 parts, phytone 5 parts, sucrose 1.2 parts, trehalose 1.5
Part, lentinan 1.6 parts, 0.5 part of magnesium sulfate, 1.2 parts of calcium chloride, sodium sulfate 0.8
Part, 0.2 part of gelatin, 1.2 parts of mannitol, polyvinylpyrrolidone 0.3 part, para arrowroot
0.8 part of powder, Pulvis Talci 0.5 part, chitin 0.2 part, potassium dihydrogen phosphate 0.3 part, phosphoric acid
Disodium hydrogen 0.2 part, the glycerol 3 parts of 40%, tween 80 0.1 part and purified water 100 parts；
Additionally, also include the aminoacid of 0.8 part, described aminoacid by weight part, including
Following components: polyglutamic acid 4 parts, aspartic acid 5 parts, phenylalanine 8 part, thread ammonia
Acid 5 parts, Cys2 part, methionine 5 parts, lysine 4 parts, leucine 0.8
Part and isoleucine 0.6 part；
4) above-mentioned phage stock solution being sub-packed in ampoule sealing, described ampoule is put into equipped with nothing
In 2 DEG C of preservations in the sealing container of water methanol.
Embodiment 3
The store method of a kind of vibrio alginolyticus phage, completes under the damp condition of 5%, bag
Include following steps:
1) vibrio alginolyticus phage solution stands 6h in 5 DEG C, flocculent deposit occurs, passes through
10000r/min is centrifuged 30min；
2) in above-mentioned precipitation, add phage buffer, by resolution of precipitate, obtain phagocytosis body fluid；
Described phage buffer by weight part, including following components: trishydroxymethylaminomethane
Hydrochlorate 12 parts, 0.1 part of sodium chloride, 3 parts of potassium chloride, 10 parts of magnesium sulfate, phosphoric acid hydrogen
Dipotassium 2 parts, sodium dihydrogen phosphate 10 parts, 0.08 part of gelatin and deionized water 100 parts, and
And use dilute hydrochloric acid and/or sodium hydroxide regulation pH to 7.2；
3) phage of phagocytosis body fluid volume 2.2 times preserves protective agent mix homogeneously and obtains phage
Stock solution, described phage preserves aminoacid described in protective agent by weight part, including following group
Point: defatted milk powder 8 parts, phytone 4 parts, sucrose 0.8 part, trehalose 0.9 part,
Lentinan 1.2 parts, 0.3 part of magnesium sulfate, 0.8 part of calcium chloride, 0.7 part of sodium sulfate,
0.1 part of gelatin, 1 part of mannitol, polyvinylpyrrolidone 0.2 part, corn starch 0.6
Part, Pulvis Talci 0.4 part, chitin 0.09 part, potassium dihydrogen phosphate 0.1 part, phosphoric acid hydrogen two
0.15 part of sodium, the glycerol 2 parts of 30%, tween 80 0.08 part and deionized water 100 parts；
Additionally, also include the aminoacid of 0.5 part, described aminoacid by weight part, including
Following components: polyglutamic acid 3 parts, aspartic acid 4 parts, phenylalanine 6 part, thread ammonia
Acid 4 parts, cysteine 0.8 part, methionine 4 parts, lysine 3 parts, leucine 0.5
Part and isoleucine 0.5 part；
4) above-mentioned phage stock solution being sub-packed in ampoule sealing, described ampoule is put into equipped with silicon
In 7 DEG C of preservations in the sealing container of glue.
Embodiment 4
The store method of a kind of lactobacillus phage, completes under the damp condition of 4%, including
Following steps:
1) lactobacillus phage solution stands 5h in 5 DEG C, flocculent deposit occurs, passes through
12000r/min is centrifuged 20min；
2) in above-mentioned precipitation, add phage buffer, by resolution of precipitate, obtain phagocytosis body fluid；
Described phage buffer by weight part, including following components: trishydroxymethylaminomethane
Hydrochlorate 8 parts, 0.2 part of sodium chloride, 3 parts of potassium chloride, 15 parts of magnesium sulfate, phosphoric acid hydrogen
Dipotassium 3 parts, sodium dihydrogen phosphate 8 parts, 0.05 part of gelatin and purified water 100 parts, and
Use dilute hydrochloric acid and/or sodium hydroxide regulation pH to 7；
3) phage of phagocytosis body fluid volume 1.5 times preserves protective agent mix homogeneously and obtains phage
Stock solution, described phage preserves aminoacid described in protective agent by weight part, including following group
Point: defatted milk powder 5 parts, phytone 5 parts, sucrose 1 part, trehalose 0.8 part,
Lentinan 1.6 parts, 0.3 part of magnesium sulfate, 0.8 part of calcium chloride, 0.8 part of sodium sulfate,
0.08 part of gelatin, 0.8 part of mannitol, polyvinylpyrrolidone 0.3 part, green starch 0.7
Part, Pulvis Talci 0.5 part, chitin 0.15 part, potassium dihydrogen phosphate 0.2 part, phosphoric acid hydrogen two
0.2 part of sodium, the glycerol 1 part of 25%, tween 80 0.06 part and purified water 100 parts；
Additionally, also include the aminoacid of 0.7 part, described aminoacid by weight part, including
Following components: polyglutamic acid 4 parts, aspartic acid 3 parts, phenylalanine 4 part, thread ammonia
Acid 3 parts, cysteine 0.9 part, methionine 3 parts, lysine 2 parts, leucine 0.8
Part and isoleucine 0.6 part；
4) above-mentioned phage stock solution being sub-packed in ampoule sealing, described ampoule is put into equipped with nothing
In 4 DEG C of preservations in the sealing container of water methanol.
Embodiment 5
The store method of a kind of phage, completes under the damp condition of 6%, including following step
Rapid:
1) phage solution stands 5h in 7 DEG C, flocculent deposit occurs, through 8000r/min
Centrifugal 20min；
2) in above-mentioned precipitation, add phage buffer, by resolution of precipitate, obtain phagocytosis body fluid；
Described phage buffer by weight part, including following components: trishydroxymethylaminomethane
Hydrochlorate 15 parts, 0.2 part of sodium chloride, 1 part of potassium chloride, 20 parts of magnesium sulfate, phosphoric acid hydrogen
Dipotassium 2 parts, sodium dihydrogen phosphate 10 parts, 0.06 part of gelatin and deionized water 100 parts, and
And use dilute hydrochloric acid and/or sodium hydroxide regulation pH to 7.5；
3) phage of phagocytosis body fluid volume 1.2 times preserves protective agent mix homogeneously and obtains phage
Stock solution, described phage preserves aminoacid described in protective agent by weight part, including following group
Point: defatted milk powder 12 parts, phytone 5 parts, sucrose 0.5 part, trehalose 0.8
Part, lentinan 1.2 parts, 0.4 part of magnesium sulfate, 0.6 part of calcium chloride, sodium sulfate 0.6
Part, 0.2 part of gelatin, 1.2 parts of mannitol, polyvinylpyrrolidone 0.2 part, ethoxy
Starch 0.7 part, Pulvis Talci 0.5 part, chitin 0.2 part, potassium dihydrogen phosphate 0.03 part,
Disodium hydrogen phosphate 0.05 part, the glycerol 3 parts of 40%, tween 80 0.1 part and deionized water
100 parts；
Additionally, also include the aminoacid of 0.4 part, described aminoacid by weight part, including
Following components: polyglutamic acid 3 parts, aspartic acid 3 parts, phenylalanine 2 part, thread ammonia
Acid 5 parts, Cys2 part, methionine 4 parts, lysine 3 parts, leucine 0.3
Part and isoleucine 0.2 part；
4) above-mentioned phage stock solution being sub-packed in ampoule sealing, described ampoule is put into equipped with nothing
In 25 DEG C of preservations in the sealing container of water calcium chloride.
After using the protectant preservation of phage 1,3,6,9 months of embodiment 1-5
Pollution of Phage situation and survival rate are investigated respectively, and as shown in table 1, phage is the most not
Pollute, and as it is shown in figure 1, phage kept the survival rate of more than 60% in 9 months.
Table 1 Pollution of Phage situation
The above is only several embodiments of invention, it is noted that for the art
Those of ordinary skill for, on the premise of without departing from inventive principle, it is also possible to make some
Improving, these improvement also should be regarded as protection scope of the present invention.
Claims (10)
1. the store method of a phage, it is characterised in that: comprise the following steps:
1) phage solution stands 4-8h in 2-7 DEG C, flocculent deposit occurs, is centrifuged 20-40min through 8000-12000r/min；
2) in above-mentioned precipitation, add phage buffer, by resolution of precipitate, obtain phagocytosis body fluid；
3) phage of phagocytosis body fluid volume 1-2.5 times preserves protective agent mix homogeneously and obtains phage stock solution, and described phage preserves aminoacid described in protective agent by weight part, including following components: defatted milk powder 5-12 part, peptone 3-5 part, sucrose
0.5-1.2 part, trehalose 0.8-1.5 part, lentinan
1-1.6 part, magnesium sulfate 0.2-0.5 part, calcium chloride 0.6-1.2 part, sodium sulfate 0.6-0.8 part, gelatin 0.05-0.2 part, mannitol 0.8-1.2 part, polyvinylpyrrolidone 0.1-0.3 part, starch 0.5-0.8 part, Pulvis Talci 0.3-0.5 part, chitin 0.08-0.2 part, potassium dihydrogen phosphate 0.03-0.3 part, disodium hydrogen phosphate 0.05-0.2 part, glycerol 1-3 part, tween 80 0.02-0.1 part and 100 parts of water；
4) above-mentioned phage stock solution is sub-packed in ampoule sealing, carries out frozen in room temperature or 2-7 DEG C.
The store method of phage the most according to claim 1; it is characterized in that: described phage preserves protective agent by weight part; also include the aminoacid of 0.3-0.8 part; described aminoacid by weight part, including following components: polyglutamic acid 1-4 part, aspartic acid 3-5 part, phenylalanine 2-8 part, serine 2-5 part, cysteine
0.5-2 part, methionine 3-5 part, lysine 2-4 part, leucine 0.3-0.8 part and isoleucine 0.2-0.6 part.
The store method of phage the most according to claim 1, it is characterised in that: described ampoule put into equipped with in the sealing container of anhydrous calcium chloride or absolute methanol or silica gel in room temperature or 2-7 DEG C of preservation.
The store method of phage the most according to claim 1, it is characterised in that: described step completes under the damp condition of 4-6%.
The store method of phage the most according to claim 1, it is characterized in that: described phage buffer by weight part, including following components: Tri(Hydroxymethyl) Amino Methane Hydrochloride 8-15 part, sodium chloride 0.05-0.2 part, potassium chloride 1-5 part, magnesium sulfate 5-20 part, dipotassium hydrogen phosphate
1-3 part, sodium dihydrogen phosphate 8-12 part, gelatin 0.05-0.1 part and 100 parts of water, and regulate pH to 7-7.5.
The store method of phage the most according to claim 1, it is characterised in that: described starch is the one of potato starch, tapioca, corn starch, green starch or hetastarch.
The store method of phage the most according to claim 1, it is characterised in that: described peptone is phytone.
The store method of phage the most according to claim 1, it is characterised in that: described glycerol is the glycerol of 20%-40%.
The store method of phage the most according to claim 1 or 5, it is characterised in that: described water is deionized water or purified water.
The preparation method of the store method of phage the most according to claim 5, it is characterised in that: use dilute hydrochloric acid and/or sodium hydrate regulator solution pH.
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Cited By (5)
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| CN109988753A (en) * | 2019-04-04 | 2019-07-09 | 中国科学院南京土壤研究所 | The freeze drying protectant and its preparation method and application of Friedlander's bacillus bacteriophage |
| CN112779163A (en) * | 2021-01-07 | 2021-05-11 | 江苏省农业科学院 | Bacteriophage normal-temperature storage material and preparation method and application thereof |
| CN113201458A (en) * | 2020-12-21 | 2021-08-03 | 青岛诺安百特生物技术有限公司 | Composition for improving stability of phage preparation and application thereof |
| CN114015758A (en) * | 2021-10-15 | 2022-02-08 | 无锡百泰克生物技术有限公司 | Freeze-drying protective agent, fluorescent PCR detection kit and freeze-drying process |
| WO2023200285A1 (en) * | 2022-04-15 | 2023-10-19 | 씨제이제일제당 (주) | Novel excipient composition for maintaining long-term distribution storage stability of bacteriophages |
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| Publication number | Priority date | Publication date | Assignee | Title |
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| CN109988753A (en) * | 2019-04-04 | 2019-07-09 | 中国科学院南京土壤研究所 | The freeze drying protectant and its preparation method and application of Friedlander's bacillus bacteriophage |
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| CN112779163A (en) * | 2021-01-07 | 2021-05-11 | 江苏省农业科学院 | Bacteriophage normal-temperature storage material and preparation method and application thereof |
| CN114015758A (en) * | 2021-10-15 | 2022-02-08 | 无锡百泰克生物技术有限公司 | Freeze-drying protective agent, fluorescent PCR detection kit and freeze-drying process |
| WO2023200285A1 (en) * | 2022-04-15 | 2023-10-19 | 씨제이제일제당 (주) | Novel excipient composition for maintaining long-term distribution storage stability of bacteriophages |
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