CN104225606A - Preparation method of novel universal vaccine or antibiotic heat-resistant protectant - Google Patents
Preparation method of novel universal vaccine or antibiotic heat-resistant protectant Download PDFInfo
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- CN104225606A CN104225606A CN201310257814.9A CN201310257814A CN104225606A CN 104225606 A CN104225606 A CN 104225606A CN 201310257814 A CN201310257814 A CN 201310257814A CN 104225606 A CN104225606 A CN 104225606A
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Abstract
The invention discloses a preparation method of a novel universal vaccine or antibiotic heat-resistant protectant. On the basis of raw materials in parts by weight, the method comprises the following steps: (1) extracting silk fibroin from silk; (2) preparing an aqueous liquid of the silk fibroin; (3) concentrating the aqueous liquid of the silk fibroin; and (4) packaging a vaccine, thereby finally obtaining the novel universal vaccine or antibiotic heat-resistant protectant. The raw materials of the novel universal vaccine or antibiotic heat-resistant protectant are vast in source, wide in production place distribution and low in obtainment cost.
Description
Technical field
The present invention is in particular to the manufacture method of a kind of novel universal type vaccine, antibiotic heat resisting protective.
Background technology
The stability of vaccine is directly connected to the size of its available protecting power, and the antibiotic antibacterial efficacy of part also can be subject to having a strong impact on of temperature.Present stage, guarantee that vaccine or the stable main method of antibiotic are cryopreservation, namely utilize low temperature to reduce vaccine or antibiotic degeneration or deactivation rate, thus improve their shelf-life.Usually have 4 DEG C and-20 DEG C of two kinds of storage temperatures, accordingly for the transport of its product and preservation and the cold chain system set up also accounted for that they (particularly vaccine) uses totle drilling cost about about 80%.But, nonetheless, still cannot ensure that vaccine or antibiotic can not reduce its stability in actual use because of the fluctuation of temperature, and finally lose its due protection.Particularly on cold chain system shortcoming or incomplete outlying underdeveloped region and other places.Therefore; how can reduce vaccine and antibiotic to the dependence of cold chain system, how protect vaccine or antibiotic when higher than a great problem keeping self stability to become Ge great drug manufacturer (particularly production of vaccine producer) when refrigerated storage temperature.
The present invention is exactly in view of the foregoing; utilize the technology such as Bio-Nano-Materials and protein molecule in world today's main flow and forward position; the contradiction between Vaccine effectiveness and temperature for above this vaccine or antibiotic; brand-new, simple low cost is proposed, and more effective from the essential aspect of biomaterial, to have more the medicinal universal heat resisting protective of development and promotion prospect method of preparation and use.Be intended to solve responsive to temperature type medicament from new angle and in use hinder its greatest problem effectively utilized, for the protectant exploitation of the universal medicament of a new generation and development provide new thinking and direction.Novel universal vaccine provided by the invention, antibiotic heat resisting protective preparation method, relate to microorganism and preserve field, its heat resisting protective prepared all has good heat-resisting protective effect to multiple vaccine and antibiotic.
This technology is in nano material based on current biological engineering, the theoretical basis that the fields such as protrinology are developed and technological means, practical problem according to society sets out, and for vaccine or antibiotic safe and effective utilization, considers what the aspect such as cost and technique proposed simultaneously.In the actual mechanical process of this technology, mainly utilize natural Bombyx bombycis as raw material, utilize the fibroin albumen composition in specific proteins extractive technique acquisition fibroin, and by the production technology effectively simplified and integrate, prepared.Raw materials technology source is wide, and cost is low.Whole preparation process is simple, and versatility is high, can be simple and quick be dissolved in existing medicine or production of vaccine technique.In the preparation process of this heat resisting protective; by the reconstruct of the fibroin to special screening; make silkworm fibroin molecule can form good space package structure; vaccine virus granule, antigen molecule, drug molecule etc. are effectively wrapped up; deaden the contact of the chemicals such as itself and extraneous various enzymes; reduce the speed being wrapped granular degeneration or decomposition; finally play protection vaccine or the antibiotic etc not effect of rapid deactivation by the rising of temperature; not only there is heat-resisting effect, also have certain slow releasing function concurrently simultaneously.Compare currently available vaccines, existing vaccines heat resisting protective, preparation technology of the present invention is more simple, cost is lower.The most important thing is, the protective agent raw material in the present invention only has a kind of composition, and whole preparation process does not use any noxious substance, safe and simple, and the fibroin albumen of parcel nontoxic, do not cause immunoreation and can 100% by protease hydrolysis.Thus also ensured this heat resisting protective absolute safety to organism in the use procedure of final products.
At present, in society, existing vaccine or antibiotic heat resisting protective about have about 20 kinds, but form according to special proportioning collocation by the organic or inorganic composition more than more than 3 kinds.Due to the use of Multiple components, the proportioning of its heat-resisting protective effect and its composition is made to have higher contact, thus production technology relative complex.Simultaneously, owing to wherein having one or Multiple components to be by chemosynthesis or multiple refine, make its use cost relatively high, and there is certain toxicity to organism in some chemical properties, its use amount and proportioning need strict control, and there is certain potential source biomolecule safety problem.Further, how many vaccines or antibiotic safe storage temperature can only be brought up to 4 DEG C greatly absolutely by the heat resisting protective existing for present stage, and the cost increase therefore produced makes its price rise about 200%.Comparatively speaking, the present invention uses natural Bombyx bombycis for raw material, and China is as sericulture big country, and raw material obtains and easily, cost is very low.And the silkworm fibroin safety non-toxic extracted by Bombyx bombycis, without anaphylaxis, 100% enzyme hydrolysis, protective agent is applied widely.Meanwhile, owing to only using a kind of raw material, production technology is simple, and be easy to integrate with existing production technology, and whole preparation process does not use noxious substance, environmental pollution is little.
Summary of the invention
The object of the invention is to overcome deficiency of the prior art, provide the manufacture method of a kind of novel universal type vaccine, antibiotic heat resisting protective, to solve the problem in background technology.
A manufacture method for novel universal type vaccine, antibiotic heat resisting protective, following raw material, according to weight ratio, comprises the following steps:
(1) extraction of silkworm fibroin: the Na2CO3 aqueous solution preparing 0.02 molar concentration, and by 100 parts of Bombyx bombycis as wherein decocting in water 20 minutes at 100 DEG C, then its abundant rinse in distilled water, with removing wherein contained sericin, is obtained fibroin albumen;
(2) preparation of silkworm fibroin aqueous solution: LiBr (lithium bromide) solution preparing 9.3 molar concentrations, and 60 DEG C by above-mentioned steps propose fibroin albumens as solution in 5 hours, fully dissolve to make fibroin albumen;
(3) silkworm fibroin aqueous solution is concentrated: preparation 25w/v%PEG solution, and dialyse corresponding with its PEG solution for the silkworm fibroin aqueous solution of acquisition or carry out chromatography, final acquisition molecular weight is greater than the silk fibroin water solution of 3.5KDa, silk fibroin water solution after concentrated is placed in 7 DEG C and saves backup, to avoid too early aggregate and precipitate;
(4) the parcel process of vaccine: vaccine is mixed with 1: 1 ratio with the silkworm fibroin aqueous solution of 8w/v%, can according to the different choice of concrete production of vaccine technique after mixing: water preparation vaccine: after mixing, mixture is left standstill 12 hours, so that fibroin albumen completes vaccine parcel; Freeze dried vaccine: after mixing, mixture is placed in freeze dryer, according to the difference of freeze dryer used, at-45 DEG C of freezing 30-480 minute, freeze-drying process P-drying:-20 DEG C, 2-40 hour; S-drying:35 DEG C, 1-25 hour, final acquisition type vaccine, antibiotic heat resisting protective.
Antibiotic parcel process: antibiotic is mixed with solution (for water-fast antibiotic, can select to dissolve with ethanol etc.), and by this antibiotic solution and silkworm fibroin aqueous solution.Concrete hybrid standard is: in final mixed solution, the ultimate density of silkworm fibroin is 6w/v%, in final mixed solution, antibiotic ultimate density is: 25mg/ml-50mg/ml, then, this mixed liquor is placed in left at room temperature 12 hours, so that fibroin albumen completes antibiotic parcel.If also undried after 12 hours, continue normal temperature drying process.For the final desciccate obtained, if antibiotic final concentration is too high, then protective agent of the present invention reduces relatively to its protected effect.
The qualification of novel universal heat resisting protective effect:
1, vaccine heat-resisting protective effect identification: the vaccine wrapped up through heat resistance protective agent is placed in respectively: deposit 6 months under 4 DEG C, 25 DEG C, 37 DEG C, 45 DEG C conditions, and within every 1 week, being carry out vaccine protection in sampling period to tire test.Corresponding often group is to deposit the identical vaccine without parcel of same time at the same temperature as experiment contrast group.
2, antibiotic heat-resisting protective effect identification: the antibiotic wrapped up through heat resistance protective agent is placed in respectively: 4 DEG C, 25 DEG C, 37 DEG C, 60 DEG C and deposit 4 weeks under having illumination condition, and 10 months.And utilize E.coli to represent as gram negative bacteria, or S.aureus represents as gram positive bacteria, carries out Drug resistance experiment to the antibiotic deposited at different temperatures.Often group is to deposit not carrying out for contrasting through the identical antibiotic of protective agent parcel of same time at the same temperature.Within every 1 week or 1 month, adopt antibiotic as a sampling period and carry out Drug resistance test.
The present invention proposes new resolving ideas and method to the heat-resistant stability of the medical product such as vaccine or antibiotic from raw material uses.By biomaterials art and protrinology technology; with modal material and the simplest technique; achieve the preparation of novel universal type heat resisting protective; eliminate existing similar technique high cost, toxic side effect existing in production process or use procedure, have the problems such as stronger stress; avoid because multicomponent uses the difficult problem of each composition proportion of balance and the effect that must consider, ensure that heat resisting protective most simplification in use.New generation vaccine/antibiotic heat resisting protective farthest can be made to give full play to its due function and efficacy in different production system, improve vaccine or antibiotic effective shelf-life, reduce its dependency to cold chain system.Make vaccine or antibiotic can play better effect in more occasion.
Tested by above-mentioned authentication method, the vaccine of the novel protective agent parcel prepared by the present invention can protect the reduction of tiring to be no more than 10% under 37 DEG C of preservation conditions after 6 months.Even can under 45 DEG C of conditions, after 6 months, still keep original protection to tire more than 85%.Fully show superior heat-resisting protective.And for antibiotic, the antibiotic of the novel protective agent parcel prepared by the present invention still can keep more than at least 95% antibacterial activity under 37 DEG C of illumination preservation conditions after 4 weeks, under 60 DEG C of preservation conditions, even after 4 weeks, still have the antibacterial activity of 80%.Again demonstrate the protectant effect of heat resistance.
The raw material used is irreplaceable, because the key core that the silkworm fibroin be rich in Bombyx bombycis has been protective agent to be prepared.Therefore to reach protected effect, Bombyx bombycis raw material and silkworm fibroin irreplaceable.But in concrete technology implementation process, production technology can change to some extent, produces the end-product with this technical products with small differences, but its thinking, principle and essence and function are constant.
Beneficial effect:
Simultaneously the novel universal type vaccine prepared in the present invention, the maximum feature of antibiotic heat resisting protective possessed following multiple advantage:
1, extensively, procurement cost is low for wide raw material source, places of origin of raw materials distribution;
2, the protectant processing technique of heat resistance is simple, power consumption of polymer processing is low, can well incorporate existing procucts production technology;
3, new protective agent does not use noxious substance in the course of processing, and environmental pollution is little;
4, new self totally nontoxic of protective agent material, to the special population such as infant, anemia of pregnant woman safety;
5, new protective agent material itself is without anaphylactogen, and human body or animal body can not be made to produce anaphylaxis;
6, new protective agent material belongs to completely biodegradable material, under the existence of protease, can complete degraded at about about 30 days;
7, new protective agent is applicable on freeze-dry process production medicament completely, and has certain frozen-dried protective effect concurrently.
Detailed description of the invention
Reaching object to make technological means of the present invention, creation characteristic, workflow, using method and effect is easy to understand, below in conjunction with tool embodiment, setting forth the present invention further.
A manufacture method for novel universal type vaccine, antibiotic heat resisting protective, following raw material, according to weight ratio, comprises the following steps:
(1) extraction of silkworm fibroin: the Na2CO3 aqueous solution preparing 0.02 molar concentration, and by 100 parts of Bombyx bombycis as wherein decocting in water 20 minutes at 100 DEG C, then its abundant rinse in distilled water, with removing wherein contained sericin, is obtained fibroin albumen;
(2) preparation of silkworm fibroin aqueous solution: LiBr (lithium bromide) solution preparing 9.3 molar concentrations, and 60 DEG C by above-mentioned steps propose fibroin albumens as solution in 5 hours, fully dissolve to make fibroin albumen;
(3) silkworm fibroin aqueous solution is concentrated: preparation 25w/v%PEG solution, and dialyse corresponding with its PEG solution for the silkworm fibroin aqueous solution of acquisition or carry out chromatography, final acquisition molecular weight is greater than the silk fibroin water solution of 3.5KDa, silk fibroin water solution after concentrated is placed in 7 DEG C and saves backup, to avoid too early aggregate and precipitate;
(4) the parcel process of vaccine: vaccine is mixed with 1: 1 ratio with the silkworm fibroin aqueous solution of 8w/v%, can according to the different choice of concrete production of vaccine technique after mixing: water preparation vaccine: after mixing, mixture is left standstill 12 hours, so that fibroin albumen completes vaccine parcel; Freeze dried vaccine: after mixing, mixture is placed in freeze dryer, according to the difference of freeze dryer used, at-45 DEG C of freezing 30-480 minute, freeze-drying process P-drying:-20 DEG C, 2-40 hour; S-drying:35 DEG C, 1-25 hour, final acquisition type vaccine, antibiotic heat resisting protective.
Antibiotic parcel process: antibiotic is mixed with solution (for water-fast antibiotic, can select to dissolve with ethanol etc.), and by this antibiotic solution and silkworm fibroin aqueous solution.Concrete hybrid standard is: in final mixed solution, the ultimate density of silkworm fibroin is 6w/v%, in final mixed solution, antibiotic ultimate density is: 25mg/ml-50mg/ml, then, this mixed liquor is placed in left at room temperature 12 hours, so that fibroin albumen completes antibiotic parcel.If also undried after 12 hours, continue normal temperature drying process.For the final desciccate obtained, if antibiotic final concentration is too high, then protective agent of the present invention reduces relatively to its protected effect.
The qualification of novel universal heat resisting protective effect:
1, vaccine heat-resisting protective effect identification: the vaccine wrapped up through heat resistance protective agent is placed in respectively: deposit 6 months under 4 DEG C, 25 DEG C, 37 DEG C, 45 DEG C conditions, and within every 1 week, being carry out vaccine protection in sampling period to tire test.Corresponding often group is to deposit the identical vaccine without parcel of same time at the same temperature as experiment contrast group.
2, antibiotic heat-resisting protective effect identification: the antibiotic wrapped up through heat resistance protective agent is placed in respectively: 4 DEG C, 25 DEG C, 37 DEG C, 60 DEG C and deposit 4 weeks under having illumination condition, and 10 months.And utilize E.coli to represent as gram negative bacteria, or S.aureus represents as gram positive bacteria, carries out Drug resistance experiment to the antibiotic deposited at different temperatures.Often group is to deposit not carrying out for contrasting through the identical antibiotic of protective agent parcel of same time at the same temperature.Within every 1 week or 1 month, adopt antibiotic as a sampling period and carry out Drug resistance test.
The present invention proposes new resolving ideas and method to the heat-resistant stability of the medical product such as vaccine or antibiotic from raw material uses.By biomaterials art and protrinology technology; with modal material and the simplest technique; achieve the preparation of novel universal type heat resisting protective; eliminate existing similar technique high cost, toxic side effect existing in production process or use procedure, have the problems such as stronger stress; avoid because multicomponent uses the difficult problem of each composition proportion of balance and the effect that must consider, ensure that heat resisting protective most simplification in use.New generation vaccine/antibiotic heat resisting protective farthest can be made to give full play to its due function and efficacy in different production system, improve vaccine or antibiotic effective shelf-life, reduce its dependency to cold chain system.Make vaccine or antibiotic can play better effect in more occasion.
Tested by above-mentioned authentication method, the vaccine of the novel protective agent parcel prepared by the present invention can protect the reduction of tiring to be no more than 10% under 37 DEG C of preservation conditions after 6 months.Even can under 45 DEG C of conditions, after 6 months, still keep original protection to tire more than 85%.Fully show superior heat-resisting protective.And for antibiotic, the antibiotic of the novel protective agent parcel prepared by the present invention still can keep more than at least 95% antibacterial activity under 37 DEG C of illumination preservation conditions after 4 weeks, under 60 DEG C of preservation conditions, even after 4 weeks, still have the antibacterial activity of 80%.Again demonstrate the protectant effect of heat resistance.
More than show and describe ultimate principle of the present invention and principal character and advantage of the present invention.The technical staff of the industry should understand; the present invention is not restricted to the described embodiments; what describe in above-described embodiment and description just illustrates principle of the present invention; without departing from the spirit and scope of the present invention; the present invention also has various changes and modifications, and these changes and improvements all fall in the claimed scope of the invention.Application claims protection domain is defined by appending claims and equivalent thereof.
Claims (1)
1. a manufacture method for novel universal type vaccine, antibiotic heat resisting protective, is characterized in that, following raw material, according to weight ratio, comprises the following steps:
(1) extraction of silkworm fibroin: the Na2CO3 aqueous solution preparing 0.02 molar concentration, and by 100 parts of Bombyx bombycis as wherein decocting in water 20 minutes at 100 DEG C, then its abundant rinse in distilled water, with removing wherein contained sericin, is obtained fibroin albumen;
(2) preparation of silkworm fibroin aqueous solution: LiBr (lithium bromide) solution preparing 9.3 molar concentrations, and 60 DEG C by above-mentioned steps propose fibroin albumens as solution in 5 hours, fully dissolve to make fibroin albumen;
(3) silkworm fibroin aqueous solution is concentrated: preparation 25w/v%PEG solution, and dialyse corresponding with its PEG solution for the silkworm fibroin aqueous solution of acquisition or carry out chromatography, final acquisition molecular weight is greater than the silk fibroin water solution of 3.5KDa, silk fibroin water solution after concentrated is placed in 7 DEG C and saves backup, to avoid too early aggregate and precipitate;
(4) the parcel process of vaccine: vaccine is mixed with 1: 1 ratio with the silkworm fibroin aqueous solution of 8w/v%, can according to the different choice of concrete production of vaccine technique after mixing: water preparation vaccine: after mixing, mixture is left standstill 12 hours, so that fibroin albumen completes vaccine parcel; Freeze dried vaccine: after mixing, mixture is placed in freeze dryer, according to the difference of freeze dryer used, at-45 DEG C of freezing 30-480 minute, freeze-drying process P-drying:-20 DEG C, 2-40 hour; S-drying:35 DEG C, 1-25 hour, final acquisition type vaccine, antibiotic heat resisting protective.
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Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN107233576A (en) * | 2017-08-08 | 2017-10-10 | 江苏省农业科学院 | Heat resisting protective, swine fever can room temperature preservation live vaccines and its preparation method and application |
| CN109030163A (en) * | 2018-06-27 | 2018-12-18 | 厦门大学 | A kind of fibroin albumen microballoon of wrapping biological sample and preparation method thereof |
| CN111729085A (en) * | 2020-07-07 | 2020-10-02 | 广东丝源集团有限公司 | Heat-resistant vaccine and preparation method thereof |
-
2013
- 2013-06-20 CN CN201310257814.9A patent/CN104225606A/en active Pending
Cited By (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN107233576A (en) * | 2017-08-08 | 2017-10-10 | 江苏省农业科学院 | Heat resisting protective, swine fever can room temperature preservation live vaccines and its preparation method and application |
| WO2019028976A1 (en) * | 2017-08-08 | 2019-02-14 | 江苏省农业科学院 | Heat-resistant protective agent, swine fever vaccine allowing for preservation at room temperature, and preparation method therefor and application thereof |
| JP2019532949A (en) * | 2017-08-08 | 2019-11-14 | 江▲蘇▼省▲農▼▲業▼科学院Ｊｉａｎｇｓｕ Ａｃａｄｅｍｙ Ｏｆ Ａｇｒｉｃｕｌｔｕｒａｌ Ｓｃｉｅｎｃｅｓ | Heat-protective agent, method for producing swine cholera live vaccine that can be stored at room temperature, and use thereof |
| CN107233576B (en) * | 2017-08-08 | 2020-02-07 | 江苏省农业科学院 | Heat-resistant protective agent, vaccine capable of preserving survival of swine fever at room temperature and preparation method and application thereof |
| CN109030163A (en) * | 2018-06-27 | 2018-12-18 | 厦门大学 | A kind of fibroin albumen microballoon of wrapping biological sample and preparation method thereof |
| CN111729085A (en) * | 2020-07-07 | 2020-10-02 | 广东丝源集团有限公司 | Heat-resistant vaccine and preparation method thereof |
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