CN102167726A - Method for separating and extracting active polypeptide and immune ribonucleic acid by utilizing blood and spleen of animal - Google Patents
Method for separating and extracting active polypeptide and immune ribonucleic acid by utilizing blood and spleen of animal Download PDFInfo
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- CN102167726A CN102167726A CN2011100411378A CN201110041137A CN102167726A CN 102167726 A CN102167726 A CN 102167726A CN 2011100411378 A CN2011100411378 A CN 2011100411378A CN 201110041137 A CN201110041137 A CN 201110041137A CN 102167726 A CN102167726 A CN 102167726A
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Abstract
The invention provides a method for separating and extracting active polypeptide and immune ribonucleic acid by utilizing the blood and spleen of an animal. The method is as follows: the advanced lysis technology is utilized, the blood and spleens of healthy livestock and poultry are used as raw materials, the fat of the raw materials is removed, cells are broken, and inactivation, freezing and thawing, centrifugation, coarse filtration, chromatographic separation, fine filtration, preparing and subpackaging are performed to prepare active polypeptide oral solution. The product has small molecular weight and does not have antigenicity at all; and after being used, the product can not stimulate the organism to generate antibody and can not be neutralized by antibody. By using the active polypeptide oral solution, the animal immunity can be enhanced and the anti-infectious capability of organism can be increased; the oral solution can not be damaged by trypsin, DNA enzyme and RNA enzyme, the oral solution has good application effect; and the oral solution has high activity, low clinical dosage, no toxic or side effect and no medicament residue, is safe to use and takes effect fast. The active polypeptide oral solution can be used to prevent and cure the diseases caused by virus infection.
Description
Technical field
The present invention relates to a kind of method of utilizing animal blood and spleen separation and Extraction active polypeptide and immune ribonucleic acid, active polypeptide and immune ribonucleic acid (iRNA) are mainly used in this zoonosis toxicity prevention and treatment of diseases.
Background technology
In recent years, along with people's growth in the living standard, fowl poultry kind food demand amount increases year by year, thereby has driven developing on an unprecedented scale of aquaculture.Along with the fast development of aquaculture, the disease of fowl poultry kind is also increasing, and individual dosage is increasing, and feeding cost improves gradually, causes culture benefit to glide.Owing to Western medicine harm and the residual food-safety problem that brings of product medicine, more and more cause showing great attention to of compatriots.So go down, will badly influence the sector and continue development healthily.In the face of the situation of this sternness, our scientific research personnel of company organization, and under relevant expert's guidance, a kind of active polypeptide preparation is produced in invention, is used for the control of fowl poultry kind Animal diseases, both drug residue free and toxic side effect did not have drug-dependent yet.
Summary of the invention
The object of the present invention is to provide a kind of method of utilizing animal blood and spleen separation and Extraction active polypeptide and immune ribonucleic acid, utilize advanced lysis technology, antigens such as various bacteriums of filtering safely and effectively and protein, extract high purity to greatest extent, highly active effective ingredient, this highly purified active polypeptide preparation keep epidemic prevention that its previous generation once was subjected to and cross the full detail of taking medicine after the disease, carry or pass to livestock and poultry individuality of future generation, individuality after the reception is promptly no longer suffered from the disease of originally having suffered from, thereby realizes the purpose of treatment and prevention.
Method of the present invention adopts following steps:
1 removes fat: get the fowl poultry kind spleen, remove the fat in the raw material;
2 weighings are cleaned: accurately weigh and remove fat back raw material, clean 2～3 times with purified water;
3 coarse reductions: spleen is mixed by corresponding proportion with blood and purified water, it was ground several minutes, make homogenate with colloidal mill; The weight ratio of spleen and purified water is 1:2, and the weight ratio of spleen and blood is 1:5;
4 micronizing: homogenate is carried out micronized pulverization through biomixer, and the solution after the fragmentation is packed in the container after the sterilization;
5 deactivations: add formaldehyde solution by 0.2% of broken liquid total amount, 36～38 ℃ of deactivations;
6 freeze thawing:, put-38 ℃ of freezers with the homogenate of deactivation and thaw after freezing;
7 centrifugations: the homogenate that will thaw is put 2 ℃, and with 3600r/min centrifugal 15 minutes, remove precipitation, it is standby to get supernatant liquor;
8 filtration chromatographies: with supernatant liquor process coarse filtration, utilize chromatography column to carry out chromatography more earlier, and collect chromatographic solution;
9 smart filters: the millipore filtration of chromatographic solution with 2 μ m filtered, and is that 12000 dalton's ultra-filtration membranes carry out the essence filter with molecular weight cut-off then;
10 preparations: the solution after the essence filter is moved in the Agitation Tank, transfer between pH to 6.8～7.5 mixing with hydrochloric acid or sodium hydroxide solution;
11 packing: quantitatively packing seals; Active polypeptide content is not less than 1.8 mg/ml, and rna content is not less than 60 mcg/ml;
12 preserve: freezing preservation under-28 ℃ of conditions.
Because the novel immunosuppressive disease of animal causes the not enough or inefficacy of traditional vaccine immune efficacy, has caused very big loss to livestock industry production.Experimental results demonstrate that active polypeptide cooperates vaccine to use the immune efficacy that can significantly improve vaccine, can improve the resistivity of animal to these immunosuppressant diseases simultaneously.Active polypeptide can also be treated animal virus and auxiliary therapy bacteriosis, strengthen the immunological competence of animal, the treatment that is used for virus disease can overcome that the conventional medicament side effect is big, the shortcoming of unsatisfactory curative effect, and the active polypeptide of utilization gene engineering method production, can reduce production costs greatly, obtain good marketability price ratio.
Spleen after utilizing live body to slaughter, blood adopt biological extraction technology to make with extra care and form as raw material.The production technique advanced person of this product, science, make finished product from being crushed to of raw material, whole technological process all is to carry out in super-clean environment and the temperature below the normal temperature, used advanced lysis technology in the technology, having reduced work in-process activated protein nucleotide chain in the multigelation process effectively is decomposed and the chance of inactivation, The Application of Technology such as improvement chromatography, filtration, antigens such as various bacteriums of filtering safely and effectively and protein extract high purity, highly active effective ingredient to greatest extent.This highly purified active polypeptide preparation keep epidemic prevention that its previous generation once was subjected to and cross the full detail of taking medicine after the disease, carry or pass to livestock and poultry individuality of future generation, individuality after the reception is promptly no longer suffered from the disease of originally having suffered from, thereby realizes the purpose of treatment and prevention.
This product has the following advantages:
1. be a kind of active polypeptide, belong to the low molecule composition of can dialysing, can be used for treatment and prevent multiple fowl poultry kind disease, and can widely apply clinically, opened up the new way of fowl poultry kind diseases prevention and treatment;
2. this product can use separately, also can use simultaneously with medicine, antibody even vaccine; Can be oral, also can inject, have characteristics easy to use, that effect is fast, and do not have any stress;
3. this product raw materials used (blood, spleen) will the severe contamination surrounding environment without handling, and turn waste into wealth after will these raw materials handling through vanguard technologies, can create great economic benefit, social benefit and environmental benefit;
4. the operation of this project will solve following outstanding problem: the residual and toxic side effect problem of common drug; The sustainable development of aquaculture; Increase added value of product, promote the development problem of recycling economy.
Embodiment
Utilize the method for animal blood and spleen separation and Extraction active polypeptide and immune ribonucleic acid below for the present invention, its step is as follows:
1 removes fat: get the fowl poultry kind spleen, remove the fat in the raw material;
2 weighings are cleaned: accurately weigh and remove fat back raw material, clean 2～3 times with purified water;
3 coarse reductions: spleen is mixed by corresponding proportion with blood and purified water, it was ground several minutes, make homogenate with colloidal mill; Spleen and purified water weight ratio are 1:2, and the weight ratio of spleen and blood is 1:5;
4 micronizing: homogenate is carried out micronized pulverization through biomixer, and the solution after the fragmentation is packed in the container after the sterilization;
5 deactivations: add formaldehyde solution (weight percent of formaldehyde solution is 35～40%), 36～38 ℃ of deactivations by 0.2% of broken liquid total amount;
6 freeze thawing:, put-38 ℃ of freezers with the homogenate of deactivation and thaw after freezing;
7 centrifugations: the homogenate that will thaw is put 2 ℃, and with 3600r/min centrifugal 15 minutes, remove precipitation, it is standby to get supernatant liquor;
8 filtration chromatographies: with supernatant liquor process coarse filtration, utilize chromatography column to carry out chromatography more earlier, and collect chromatographic solution; The chromatography column major technique is:
9 smart filters: the millipore filtration of chromatographic solution with 2 μ m filtered, and is that 12000 dalton's ultra-filtration membranes carry out the essence filter with molecular weight cut-off then;
10 preparations: the solution after the essence filter is moved in the Agitation Tank, transfer between pH to 6.8～7.5 mixing with hydrochloric acid or sodium hydroxide solution;
11 packing: quantitatively packing seals;
12 preserve: freezing preservation under-28 ℃ of conditions.
The concentration of product: polypeptide is not less than 1.8 mg/ml, and Yeast Nucleic Acid is not less than 80 mcg/ml.
Claims (1)
1. method of utilizing animal blood and spleen separation and Extraction active polypeptide and immune ribonucleic acid is characterized in that adopting following steps:
(1) removes fat: get the fowl poultry kind spleen, remove the fat in the raw material;
(2) weighing is cleaned: accurately weigh and remove fat back raw material, clean 2～3 times with purified water;
(3) coarse reduction: spleen is mixed by corresponding proportion with blood and purified water, it was ground several minutes, make homogenate with colloidal mill; The weight ratio of spleen and purified water is 1:2, and the weight ratio of spleen and blood is 1:5;
(4) micronizing: homogenate is carried out micronized pulverization through biomixer, and the solution after the fragmentation is packed in the container after the sterilization;
(5) deactivation: add formaldehyde solution by 0.2% of broken liquid total amount, 36～38 ℃ of deactivations;
(6) freeze thawing:, put-38 ℃ of freezers with the homogenate of deactivation and thaw after freezing;
(7) centrifugation: the homogenate that will thaw is put 2 ℃, and with 3600r/min centrifugal 15 minutes, remove precipitation, it is standby to get supernatant liquor;
(8) filtration chromatography: with supernatant liquor process coarse filtration, utilize chromatography column to carry out chromatography more earlier, and collect chromatographic solution;
(9) smart filter: the millipore filtration of chromatographic solution with 2 μ m filtered, and is that 12000 dalton's ultra-filtration membranes carry out the essence filter with molecular weight cut-off then;
(10) preparation: the solution after the essence filter is moved in the Agitation Tank, transfer between pH to 6.8～7.5 mixing with hydrochloric acid or sodium hydroxide solution;
(11) packing: quantitatively packing seals; Active polypeptide content is not less than 1.8 mg/ml, and rna content is not less than 60 mcg/ml;
(12) preserve: freezing preservation under-28 ℃ of conditions.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN2011100411378A CN102167726A (en) | 2011-02-21 | 2011-02-21 | Method for separating and extracting active polypeptide and immune ribonucleic acid by utilizing blood and spleen of animal |
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| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN2011100411378A CN102167726A (en) | 2011-02-21 | 2011-02-21 | Method for separating and extracting active polypeptide and immune ribonucleic acid by utilizing blood and spleen of animal |
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| CN102167726A true CN102167726A (en) | 2011-08-31 |
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| CN2011100411378A Pending CN102167726A (en) | 2011-02-21 | 2011-02-21 | Method for separating and extracting active polypeptide and immune ribonucleic acid by utilizing blood and spleen of animal |
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Cited By (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104352521A (en) * | 2014-11-05 | 2015-02-18 | 吉林大学 | Method for preparing calf spleen extract and application in anti-tumor and immune adjustment |
| CN104352520A (en) * | 2014-11-05 | 2015-02-18 | 吉林大学 | Calf spleen extractive and application thereof to idiopathic thrombocytopenic purpura treatment |
| CN105613936A (en) * | 2016-03-24 | 2016-06-01 | 刘冬明 | Preparation method for spleen polypeptides |
| CN108129549A (en) * | 2017-12-19 | 2018-06-08 | 浙江丰安生物制药有限公司 | The extracting method of polypeptide in a kind of spleen aminopeptide |
| CN108148111A (en) * | 2017-12-19 | 2018-06-12 | 浙江丰安生物制药有限公司 | The extracting method of polypeptide in a kind of spleen aminopeptide |
| WO2022179551A1 (en) * | 2021-02-26 | 2022-09-01 | 重庆誉颜制药有限公司 | Cell lysis buffer separation apparatus for preparing polypeptide, and system and application thereof |
-
2011
- 2011-02-21 CN CN2011100411378A patent/CN102167726A/en active Pending
Cited By (10)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104352521A (en) * | 2014-11-05 | 2015-02-18 | 吉林大学 | Method for preparing calf spleen extract and application in anti-tumor and immune adjustment |
| CN104352520A (en) * | 2014-11-05 | 2015-02-18 | 吉林大学 | Calf spleen extractive and application thereof to idiopathic thrombocytopenic purpura treatment |
| CN104352521B (en) * | 2014-11-05 | 2017-12-22 | 吉林大学 | Prepare the method for calf spleen extract and the application in antitumor and immunological regulation |
| CN104352520B (en) * | 2014-11-05 | 2018-05-22 | 吉林大学 | Calf spleen extract and the purposes in the treatment of primary immune thrombocytopenia |
| CN105613936A (en) * | 2016-03-24 | 2016-06-01 | 刘冬明 | Preparation method for spleen polypeptides |
| CN108129549A (en) * | 2017-12-19 | 2018-06-08 | 浙江丰安生物制药有限公司 | The extracting method of polypeptide in a kind of spleen aminopeptide |
| CN108148111A (en) * | 2017-12-19 | 2018-06-12 | 浙江丰安生物制药有限公司 | The extracting method of polypeptide in a kind of spleen aminopeptide |
| CN108129549B (en) * | 2017-12-19 | 2020-06-05 | 浙江丰安生物制药有限公司 | Method for extracting polypeptide in spleen aminopeptide |
| CN108148111B (en) * | 2017-12-19 | 2020-06-05 | 浙江丰安生物制药有限公司 | Method for extracting polypeptide in spleen aminopeptide |
| WO2022179551A1 (en) * | 2021-02-26 | 2022-09-01 | 重庆誉颜制药有限公司 | Cell lysis buffer separation apparatus for preparing polypeptide, and system and application thereof |
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Application publication date: 20110831 |