CN102229670A - Method for extracting immune globulin from yak blood - Google Patents
Method for extracting immune globulin from yak blood Download PDFInfo
- Publication number
- CN102229670A CN102229670A CN2011101193811A CN201110119381A CN102229670A CN 102229670 A CN102229670 A CN 102229670A CN 2011101193811 A CN2011101193811 A CN 2011101193811A CN 201110119381 A CN201110119381 A CN 201110119381A CN 102229670 A CN102229670 A CN 102229670A
- Authority
- CN
- China
- Prior art keywords
- blood
- yak
- immunoglobulin
- extracting
- yak blood
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Pending
Links
Abstract
The invention relates to a method for extracting immune globulin from yak blood. The method comprises the following steps: adding anticoagulant into fresh yak blood and then centrifuging the mixture, thereby acquiring blood plasma; utilizing an ultra-filtration membrane to purify and concentrate the blood plasma; ultrasonically extracting an interception phase with ethanol; and freeze-drying, thereby acquiring the immune globulin. The method provided by the invention is simple, high-speed, safe, pollution-free, low-cost, and low-carbon.
Description
Technical field:
The invention belongs to natural drug extractive technique field, relate to a kind of method of from yak blood, extracting immunoglobulin (Ig).
Background technology:
Immunoglobulin (Ig) (Immunoglobulin, be called for short Ig) is meant to have antibody activity, can with the sphaeroprotein of corresponding antigen generation specificity association reaction, be prevalent in mammiferous blood, tissue juice, lymph liquid and the exocrine secretion.Immunoglobulin (Ig) has important immunity and physiological regulatory action in animal body, is one of the most key component of animal vivo immuning system.Immunoglobulin (Ig) can be divided into IgG, IgA, IgM, IgD, IgE five classes.The main component of human serum immunoglobulin (Ig) is IgG, and it accounts for the 70-75% of total immunoglobulin (Ig), and molecular weight is about 150,000, contains sugar 2～3%.Although immunoglobulin (Ig) is ever-changing, similar structure is arranged all.Antibody molecule is made up of two pairs of different polypeptide chains of length, and four chains constitute Y type basic structure (H2L2) by interchain disulfide bond.The human serum immunoglobulin IgG is the most lasting in the primary immune response, most important antibody, and it only exists with monomeric form.Be that germ resistance, toxin immunity and antiviral antibody belong to IgG mostly, it plays main force's effect in anti-infective, it can promote the phagolysis (opsonization) of mononuclear macrophage, and bacteriotoxic toxicity that neutralizes (neutralizing a toxin) and virus antigen are in conjunction with making virus lose the ability (neutralization virus) of host cells infected.Immunoglobulin (Ig) generally extracts with cow's milk and pig blood.In recent years, immunoglobulin (Ig) is widely used in the development of new functional food additives, in newborn animal feed and biological new drug and the medical biochemical diagnosis, has become the important materials of department such as research and commerce, and prospect is very wide.Yet immunoglobulin (Ig) makes its product development be very restricted to the unstable of heat.
Yak is the distinctive Highland cattle kind of China, is called as " yak ", and it is abominable but the choice of nonpollution environment has very strong immunizing power to be subjected to the plateau.Again because be subjected to the irradiation of plateau intensive ultraviolet, so yak is natural aseptic resource treasure-house.Its blood did not often obtain utilizing after yak was slaughtered and gets meat, all as sewage discharge, the value of yak can't be made full use of, and increased environmental pressure.
From blood, extract immunoglobulin (Ig), method commonly used has salting-out process (as the sodium polyphosphate flocculence, the sulfate of ammoniac salting-out process), organic solvent precipitation method (as cold ethanol partition method, the sad precipitator method), the organic polymer precipitator method, the sex change precipitator method, Mierocrystalline cellulose chromatography method, gel chromatography, ultrafiltration process etc., many scholars did some deep researchs to extracting immunoglobulin (Ig), but relatively more of the integrated approach of antagonist purifying, restriction purifying purity and yield bottleneck stage---the stage of slightly carrying is then studied less.
Summary of the invention
Purpose of the present invention is exactly the deficiency at above-mentioned existence, and a kind of method of extracting immunoglobulin (Ig) from yak blood is provided.
The present invention says that the technical scheme that provides is:
1) centrifugation: in fresh yak blood, add antithrombotics, centrifugal, keep blood plasma;
2) ultrafiltration: will concentrate by ultra-filtration membrane behind the blood plasma dilute with water, washing impurity-removing must be held back phase;
3) supersound extraction: will hold back and be added in the ethanol supersound extraction and repeat united extraction liquid 2 times;
4) precipitation: with the extracting solution acid adjustment, centrifugal must the precipitation;
5) lyophilize: will precipitate lyophilize promptly.
Antithrombotics in the described step 1) is a Trisodium Citrate, and consumption is 0.3% (V/V).
Described step 2) ultra-filtration membrane in is the multiporous fiber ultra-filtration membrane of molecular weight cut-off 3000～10000D.
Extraction solvent load in the described step 3) is that 10-15 doubly measures (V/W), and extraction time is 20-30min, ambient operation.
Acid in the described step 4) is one of hydrochloric acid or sulfuric acid, and the pH after the acidifying is 7.3-7.7.
In sum, there is following advantage in the present invention:
1) is raw material with yak blood, turns waste into wealth, solved the problem that stays after yak is slaughtered;
2) solvent is simple and easy to, and toxicity is little, so aftertreatment is simple, and cost is low, and Product Safety is higher;
3) membrane sepn only needs impellent, does not need to evaporate heat dissipation, so energy consumption is little, low carbonization;
4) supersound extraction speed is than general solvent extraction efficient height, and it is low to extract temperature, and product is lost because of decomposes, also can not introduce other pollutents;
5) immunoglobulin (Ig) to separate out pH near from neutrality, therefore sour consumption is little;
6) each step process amount is big, and speed is fast, is fit to scale operation.
Further specify the present invention below in conjunction with embodiment, but the scope of protection of present invention is not limited to following embodiment.
Embodiment
Embodiment 1
Add 0.3% Trisodium Citrate in the fresh yak blood of 100L, high speed centrifugation makes the blood layering, keeps blood plasma, and the multiporous fiber ultra-filtration membrane by molecular weight cut-off 3000～10000D concentrates, and adds water washing, gets and holds back phase.Hold back mutually with 10 times of amounts (V/W) ethanol ultrasonic extraction 20min, repeat 2 times, united extraction liquid, adding hydrochloric acid, to reconcile pH be 7.3-7.7, centrifugal must the precipitation, lyophilize promptly gets immunoglobulin (Ig), and purity is 78.7%.
Embodiment 2
Add 0.3% Trisodium Citrate in the fresh yak blood of 100L, high speed centrifugation makes the blood layering, keeps blood plasma, and the multiporous fiber ultra-filtration membrane by molecular weight cut-off 3000～10000D concentrates, and adds water washing, gets and holds back phase.Hold back mutually with 15 times of amounts (V/W) ethanol ultrasonic extraction 30min, repeat 2 times, united extraction liquid, adding hydrochloric acid, to reconcile pH be 7.3-7.7, centrifugal must the precipitation, lyophilize promptly gets immunoglobulin (Ig), and purity is 78.4%.
Embodiment 3
Add 0.3% Trisodium Citrate in the fresh yak blood of 100L, high speed centrifugation makes the blood layering, keeps blood plasma, and the multiporous fiber ultra-filtration membrane by molecular weight cut-off 3000～10000D concentrates, and adds water washing, gets and holds back phase.Hold back mutually with 10 times of amounts (V/W) ethanol ultrasonic extraction 20min, repeat 2 times, united extraction liquid, adding hydrochloric acid, to reconcile pH be 7.3-7.7, centrifugal must the precipitation, lyophilize promptly gets immunoglobulin (Ig), and purity is 79.1%.
Embodiment 4
Add 0.3% Trisodium Citrate in the fresh yak blood of 100L, high speed centrifugation makes the blood layering, keeps blood plasma, and the multiporous fiber ultra-filtration membrane by molecular weight cut-off 3000～10000D concentrates, and adds water washing, gets and holds back phase.Hold back mutually with 15 times of amounts (V/W) ethanol ultrasonic extraction 30min, repeat 2 times, united extraction liquid, adding hydrochloric acid, to reconcile pH be 7.3-7.7, centrifugal must the precipitation, lyophilize promptly gets immunoglobulin (Ig), and purity is 77.9%.
Embodiment 5
Add 0.3% Trisodium Citrate in the fresh yak blood of 100L, high speed centrifugation makes the blood layering, keeps blood plasma, and the multiporous fiber ultra-filtration membrane by molecular weight cut-off 3000～10000D concentrates, and adds water washing, gets and holds back phase.Hold back mutually with 12 times of amounts (V/W) ethanol ultrasonic extraction 25min, repeat 2 times, united extraction liquid, adding hydrochloric acid, to reconcile pH be 7.3-7.7, centrifugal must the precipitation, lyophilize promptly gets immunoglobulin (Ig), and purity is 79.5%.
Embodiment 6
Add 0.3% Trisodium Citrate in the fresh yak blood of 100L, high speed centrifugation makes the blood layering, keeps blood plasma, and the multiporous fiber ultra-filtration membrane by molecular weight cut-off 3000～10000D concentrates, and adds water washing, gets and holds back phase.Hold back mutually with 15 times of amounts (V/W) ethanol ultrasonic extraction 20min, repeat 2 times, united extraction liquid, adding hydrochloric acid, to reconcile pH be 7.3-7.7, centrifugal must the precipitation, lyophilize promptly gets immunoglobulin (Ig), and purity is 77.4%.
Claims (5)
1. a method of extracting immunoglobulin (Ig) from yak blood is characterized in that described method comprises the following steps:
1) centrifugation: in fresh yak blood, add antithrombotics, centrifugal, keep blood plasma;
2) ultrafiltration: will concentrate by ultra-filtration membrane behind the blood plasma dilute with water, washing impurity-removing must be held back phase;
3) supersound extraction: will hold back and be added in the ethanol supersound extraction and repeat united extraction liquid 2 times;
4) precipitation: with the extracting solution acid adjustment, centrifugal must the precipitation;
5) lyophilize: will precipitate lyophilize promptly.
2. a method of extracting immunoglobulin (Ig) from yak blood is characterized in that the antithrombotics in the described step 1) is a Trisodium Citrate, and consumption is 0.3% (V/V).
3. a method of extracting immunoglobulin (Ig) from yak blood is characterized in that described step 2) in ultra-filtration membrane be the multiporous fiber ultra-filtration membrane of molecular weight cut-off 3000～10000D.
4. a method of extracting immunoglobulin (Ig) from yak blood is characterized in that the extraction solvent load in the described step 3) is that 10-15 doubly measures (V/W), and extraction time is 20-30min, ambient operation.
5. a method of extracting immunoglobulin (Ig) from yak blood is characterized in that the acid in the described step 4) is one of hydrochloric acid or sulfuric acid, and the pH after the acidifying is 7.3-7.7.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN2011101193811A CN102229670A (en) | 2011-05-10 | 2011-05-10 | Method for extracting immune globulin from yak blood |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN2011101193811A CN102229670A (en) | 2011-05-10 | 2011-05-10 | Method for extracting immune globulin from yak blood |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN102229670A true CN102229670A (en) | 2011-11-02 |
Family
ID=44842283
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN2011101193811A Pending CN102229670A (en) | 2011-05-10 | 2011-05-10 | Method for extracting immune globulin from yak blood |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN102229670A (en) |
Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN112661840A (en) * | 2019-12-19 | 2021-04-16 | 山东省阳信县福安清真肉类有限公司 | Process for preparing immune globulin by using bovine blood through biological method |
| CN114431419A (en) * | 2022-01-26 | 2022-05-06 | 郓城铸源生物科技有限公司 | Food raw material and preparation method thereof |
| US11604026B2 (en) | 2019-03-14 | 2023-03-14 | Terumo Bct Biotechnologies, Llc | Lyophilization loading tray assembly and system |
| US11634257B2 (en) | 2017-10-09 | 2023-04-25 | Terumo Bct Biotechnologies, Llc | Lyophilization container and method of using same |
-
2011
- 2011-05-10 CN CN2011101193811A patent/CN102229670A/en active Pending
Cited By (9)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US11634257B2 (en) | 2017-10-09 | 2023-04-25 | Terumo Bct Biotechnologies, Llc | Lyophilization container and method of using same |
| US11604026B2 (en) | 2019-03-14 | 2023-03-14 | Terumo Bct Biotechnologies, Llc | Lyophilization loading tray assembly and system |
| US11609043B2 (en) | 2019-03-14 | 2023-03-21 | Terumo Bct Biotechnologies, Llc | Lyophilization container fill fixture, system and method of use |
| US11609042B2 (en) | 2019-03-14 | 2023-03-21 | Terumo Bct Biotechnologies, Llc | Multi-part lyophilization container and method of use |
| US11740019B2 (en) | 2019-03-14 | 2023-08-29 | Terumo Bct Biotechnologies, Llc | Lyophilization loading tray assembly and system |
| US11747082B2 (en) | 2019-03-14 | 2023-09-05 | Terumo Bct Biotechnologies, Llc | Multi-part lyophilization container and method of use |
| US11815311B2 (en) | 2019-03-14 | 2023-11-14 | Terumo Bct Biotechnologies, Llc | Lyophilization container fill fixture, system and method of use |
| CN112661840A (en) * | 2019-12-19 | 2021-04-16 | 山东省阳信县福安清真肉类有限公司 | Process for preparing immune globulin by using bovine blood through biological method |
| CN114431419A (en) * | 2022-01-26 | 2022-05-06 | 郓城铸源生物科技有限公司 | Food raw material and preparation method thereof |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| Qin et al. | Selenylation modification can enhance immune-enhancing activity of Chinese angelica polysaccharide | |
| CN104498570B (en) | Ocean fish protein peptide and preparation method thereof | |
| CN102229670A (en) | Method for extracting immune globulin from yak blood | |
| CN103694348A (en) | Preparation and application method for treating swine viral diarrhea biological agent | |
| CN103190621A (en) | Propolis soft capsule and preparation method thereof | |
| CN104292352B (en) | A kind of Cortex Eucommiae fine powder, eucommia bark polycose and gutta-percha coproduction extraction separation method | |
| CN102268101A (en) | Enzyme-assisted extraction method of high-purity porphyra polysaccharide | |
| CN104498564A (en) | Low molecular weight chondroitin sulfate preparation method | |
| CN102586376B (en) | Albumen antioxidant peptide and preparation method thereof | |
| CN103356723B (en) | Cooling paste for children and preparation method thereof | |
| CN102718882B (en) | Selenylation modification method for improving immunity-enhancing activity of angelica polysaccharide | |
| CN101690813B (en) | Concentrated freeze-dried yolk antibody composite preparation for Newcastle disease and preparation process thereof | |
| CN104277135A (en) | Pleurotus tuber-regium polysaccharide and extraction method thereof | |
| CN100586454C (en) | Chinese medicine composition for birds, preparing method and application of the same | |
| CN101766252A (en) | Sheep blood high-activity immune globulin concentrate and preparation method thereof | |
| CN101824100A (en) | Heparin-12-sugar, preparation method thereof and application thereof in resisting vascular smooth muscle cell proliferation | |
| CN102898540A (en) | Extraction technology of hyaluronic acid | |
| CN101768582B (en) | Production process for modifying SOD | |
| CN103393710B (en) | A kind of Preparation method and use of fermented Cordyceps powder polysaccharides compound | |
| CN101899120A (en) | Method for refining jujube polysaccharide | |
| CN103446182B (en) | The preparation method of high-pathogenicity porcine reproductive and respiratory syndrome specific transfer factor | |
| CN102809653A (en) | Preparation and application of ELISA (Enzyme-Linked Immunosorbent Assay) kit for detecting novel bunyavirus antigen | |
| CN103349782B (en) | The preparation technology of Eimeria Tenella Yolk antibody target slow-release preparation | |
| CN102212129A (en) | Method for extracting human fibrinogen from component I through column chromatography | |
| CN104873971A (en) | Diluent for hog cholera live vaccine (spleen and lymph tissue origin) |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| C06 | Publication | ||
| PB01 | Publication | ||
| C02 | Deemed withdrawal of patent application after publication (patent law 2001) | ||
| WD01 | Invention patent application deemed withdrawn after publication |
Application publication date: 20111102 |