CN1074709A - Method for preparing thrombin - Google Patents
Method for preparing thrombin Download PDFInfo
- Publication number
- CN1074709A CN1074709A CN 92112188 CN92112188A CN1074709A CN 1074709 A CN1074709 A CN 1074709A CN 92112188 CN92112188 CN 92112188 CN 92112188 A CN92112188 A CN 92112188A CN 1074709 A CN1074709 A CN 1074709A
- Authority
- CN
- China
- Prior art keywords
- zymoplasm
- value
- damping fluid
- throw out
- making method
- 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
- 238000000034 method Methods 0.000 title claims abstract description 16
- 108090000190 Thrombin Proteins 0.000 title abstract 2
- 229960004072 thrombin Drugs 0.000 title abstract 2
- 238000004587 chromatography analysis Methods 0.000 claims abstract description 12
- 239000007788 liquid Substances 0.000 claims abstract description 8
- 210000004369 blood Anatomy 0.000 claims abstract description 6
- 239000008280 blood Substances 0.000 claims abstract description 6
- 230000001105 regulatory effect Effects 0.000 claims abstract description 3
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 claims description 11
- 238000013016 damping Methods 0.000 claims description 11
- 239000012530 fluid Substances 0.000 claims description 11
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims description 7
- 238000004440 column chromatography Methods 0.000 claims description 6
- 108010064129 Thrombogen Proteins 0.000 claims description 4
- 230000004913 activation Effects 0.000 claims description 4
- 238000005119 centrifugation Methods 0.000 claims description 4
- 229920000936 Agarose Polymers 0.000 claims description 3
- 230000002785 anti-thrombosis Effects 0.000 claims description 3
- 229960004676 antithrombotic agent Drugs 0.000 claims description 3
- 229920002307 Dextran Polymers 0.000 claims description 2
- 230000008901 benefit Effects 0.000 abstract description 5
- 230000000694 effects Effects 0.000 abstract description 2
- 239000003146 anticoagulant agent Substances 0.000 abstract 1
- 229940127219 anticoagulant drug Drugs 0.000 abstract 1
- 230000003139 buffering effect Effects 0.000 abstract 1
- 230000008021 deposition Effects 0.000 abstract 1
- 238000004519 manufacturing process Methods 0.000 abstract 1
- 108090000623 proteins and genes Proteins 0.000 abstract 1
- 102000004169 proteins and genes Human genes 0.000 abstract 1
- 208000032843 Hemorrhage Diseases 0.000 description 5
- 229960004756 ethanol Drugs 0.000 description 3
- 238000000926 separation method Methods 0.000 description 3
- 108010017384 Blood Proteins Proteins 0.000 description 2
- 102000004506 Blood Proteins Human genes 0.000 description 2
- TZCXTZWJZNENPQ-UHFFFAOYSA-L barium sulfate Chemical compound [Ba+2].[O-]S([O-])(=O)=O TZCXTZWJZNENPQ-UHFFFAOYSA-L 0.000 description 2
- 230000000740 bleeding effect Effects 0.000 description 2
- 238000004090 dissolution Methods 0.000 description 2
- 229940079593 drug Drugs 0.000 description 2
- 239000003814 drug Substances 0.000 description 2
- 230000023597 hemostasis Effects 0.000 description 2
- 230000002439 hemostatic effect Effects 0.000 description 2
- 210000002381 plasma Anatomy 0.000 description 2
- 238000001556 precipitation Methods 0.000 description 2
- 101100312904 Arabidopsis thaliana ADA2A gene Proteins 0.000 description 1
- BHPQYMZQTOCNFJ-UHFFFAOYSA-N Calcium cation Chemical compound [Ca+2] BHPQYMZQTOCNFJ-UHFFFAOYSA-N 0.000 description 1
- 102000010911 Enzyme Precursors Human genes 0.000 description 1
- 108010062466 Enzyme Precursors Proteins 0.000 description 1
- 102000009123 Fibrin Human genes 0.000 description 1
- 108010073385 Fibrin Proteins 0.000 description 1
- BWGVNKXGVNDBDI-UHFFFAOYSA-N Fibrin monomer Chemical compound CNC(=O)CNC(=O)CN BWGVNKXGVNDBDI-UHFFFAOYSA-N 0.000 description 1
- 102000008946 Fibrinogen Human genes 0.000 description 1
- 108010049003 Fibrinogen Proteins 0.000 description 1
- 208000007536 Thrombosis Diseases 0.000 description 1
- 238000010521 absorption reaction Methods 0.000 description 1
- 239000002253 acid Substances 0.000 description 1
- WNROFYMDJYEPJX-UHFFFAOYSA-K aluminium hydroxide Chemical compound [OH-].[OH-].[OH-].[Al+3] WNROFYMDJYEPJX-UHFFFAOYSA-K 0.000 description 1
- 229910021502 aluminium hydroxide Inorganic materials 0.000 description 1
- 230000023555 blood coagulation Effects 0.000 description 1
- 210000001772 blood platelet Anatomy 0.000 description 1
- 229910001424 calcium ion Inorganic materials 0.000 description 1
- 239000001506 calcium phosphate Substances 0.000 description 1
- 229910000389 calcium phosphate Inorganic materials 0.000 description 1
- 235000011010 calcium phosphates Nutrition 0.000 description 1
- 230000007812 deficiency Effects 0.000 description 1
- 229960000935 dehydrated alcohol Drugs 0.000 description 1
- 229950003499 fibrin Drugs 0.000 description 1
- 229940012952 fibrinogen Drugs 0.000 description 1
- 238000004108 freeze drying Methods 0.000 description 1
- 238000007710 freezing Methods 0.000 description 1
- 230000000025 haemostatic effect Effects 0.000 description 1
- 125000002887 hydroxy group Chemical group [H]O* 0.000 description 1
- 231100000957 no side effect Toxicity 0.000 description 1
- 239000010695 polyglycol Substances 0.000 description 1
- 229920000151 polyglycol Polymers 0.000 description 1
- 229920000642 polymer Polymers 0.000 description 1
- 238000002360 preparation method Methods 0.000 description 1
- 238000000746 purification Methods 0.000 description 1
- 238000007670 refining Methods 0.000 description 1
- 239000001509 sodium citrate Substances 0.000 description 1
- 239000001488 sodium phosphate Substances 0.000 description 1
- 229910000162 sodium phosphate Inorganic materials 0.000 description 1
- QORWJWZARLRLPR-UHFFFAOYSA-H tricalcium bis(phosphate) Chemical compound [Ca+2].[Ca+2].[Ca+2].[O-]P([O-])([O-])=O.[O-]P([O-])([O-])=O QORWJWZARLRLPR-UHFFFAOYSA-H 0.000 description 1
- HRXKRNGNAMMEHJ-UHFFFAOYSA-K trisodium citrate Chemical compound [Na+].[Na+].[Na+].[O-]C(=O)CC(O)(CC([O-])=O)C([O-])=O HRXKRNGNAMMEHJ-UHFFFAOYSA-K 0.000 description 1
- 229940038773 trisodium citrate Drugs 0.000 description 1
- RYFMWSXOAZQYPI-UHFFFAOYSA-K trisodium phosphate Chemical compound [Na+].[Na+].[Na+].[O-]P([O-])([O-])=O RYFMWSXOAZQYPI-UHFFFAOYSA-K 0.000 description 1
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- Pharmaceuticals Containing Other Organic And Inorganic Compounds (AREA)
- Polysaccharides And Polysaccharide Derivatives (AREA)
Abstract
A process for preparing thrombin from animal blood includes such steps as adding anticoagulant, chromatography, buffering liquid, regulating pH value to dewater protein molecules, and deposition at isoelectric point. The invention has the following advantages: simple process flow, small investment, low cost, short time, high purity, high activity, more benefits and the like.
Description
The present invention relates to a kind of hemostatic drug, particularly to the various hemorrhage making method that can both reach a kind of zymoplasm of fast-acting nemostatic.
At present, hemorrhage what adopt is that bleeding stopping period is long with the hemostasis of conventional hemostatic drug to various clinically, has again to pay to act on, and particularly to hemorrhage due to the hemopathy, haemostatic effect is poorer; And the use zymoplasm, bleeding stopping period is short, and effect is rapidly, and is evident in efficacy, easy to use, has no side effect.Zymoplasm directly acts on blood coagulation, makes Fibrinogen change fibrin polymer rapidly into, comprises that simultaneously red corpuscle, white corpuscle, thrombocyte, blood plasma constitute blood clot, stops up the blutpunkte, reaches the hemostasis purpose.
Cohn has set up cold ethanol method in 1940, this method master usefulness is the change (five change systems) by five factors, and plasma proteins is separated.European patent 56629, German Patent 2902657 and United States Patent (USP) 92626 are introduced respectively is to be preparation with barium sulfate, aluminium hydroxide, calcium phosphate, sodium phosphate, polyglycol and hydroxy alkanoic acid, carry out absorption, separation with calcium ion, reach the purpose of the separation of plasma proteins and purification, adopt the weak point of the zymoplasm that above-mentioned several method manufactures to be: the technical process complexity, investment is big, the cost height, and the time is long, purity is low, active low, get profit few.
The object of the invention is to avoid above-mentioned the deficiencies in the prior art part, provides a kind of technical process simple, invests for a short time, and cost is low, and the time is short, and the purity height is active high, the making method of fast-acting nemostatic medicine-zymoplasm of benefit.
Purpose of the present invention can reach by following measure: a kind of making method of zymoplasm is characterized in that: after in animal blood, adding antithrombotics, add ethanol and damping fluid again, and the PH adjustable value, centrifugation has throw out to produce (being thrombogen); The throw out that is dissolved in water, last chromatography column chromatography, it is centrifugal that effluent liquid is regulated pH value with damping fluid, has throw out to produce; The throw out that is dissolved in water again, normal temperature activation were down transferred pH value with damping fluid more than one hour, last chromatography column chromatography, and elutriant is zymoplasm.
Purpose of the present invention can also reach by following measure: after adding ethanol or damping fluid, transferring pH value is 5.3 o'clock, can from separation; Last heparin-agarose chromatography column chromatography, effluent liquid with damping fluid transfer pH value be 4.4 o'clock centrifugal; Normal temperature is the solution of activation more than 3 hours down, uses CaHPO
4Transfer H7.2, last dextran G-25 chromatography.
The present invention is described in further detail below with reference to (accompanying drawing) embodiment:
Get animal blood (as pig blood or ox blood) liquid, add the anti-freezing of antithrombotics Trisodium Citrate; The dehydrated alcohol of adding 10% in the blood plasma after the centrifugation, centrifugal when transferring PH5.3 with damping fluid HAC10%, throw out produces, and is thrombogen; The thrombogen precipitation adds water dissolution, last heparin-agarose chromatography column, and the control certain flow, centrifugal after the effusive liquid of chromatography column is transferred PH4.4 with HAC, be precipitated as purified and treat proenzyme with fixed attention; Precipitation adds 10 times with the upper volume water dissolution, and water at normal temperature bathization 1 hour is used CaHPO
4Transfer PH7.2, add CaCl
2, go up the G-25 chromatography then, effluent liquid is refining zymoplasm, and is aseptic subpackaged through degerming, is finished product after the freeze-drying.
The present invention is prior art relatively, and have following advantage: technological process is simple, invest littlely, and cost is low, and the time is short, the purity height, and active height gets benefit etc.
Claims (4)
1, a kind of making method of zymoplasm is characterized in that:
A. after in animal blood, adding antithrombotics, add ethanol and damping fluid again, the PH adjustable value, centrifugation has throw out to produce (being thrombogen);
B. the throw out that is dissolved in water, last chromatography column chromatography, it is centrifugal that effluent liquid is regulated pH value with damping fluid, has throw out to produce;
C. the throw out that is dissolved in water again, normal temperature activation were down transferred pH value with damping fluid more than one hour, last chromatography column chromatography, and elutriant is zymoplasm.
2, the making method of zymoplasm according to claim 1 is characterized in that: after adding ethanol or damping fluid, transferring pH value is 5.3 o'clock, i.e. centrifugation.
3, the making method of zymoplasm according to claim 1 is characterized in that: go up heparin-agarose chromatography column chromatography, effluent liquid with damping fluid transfer pH value be 4.4 o'clock centrifugal.
4, the making method of zymoplasm according to claim 1 is characterized in that: normal temperature is the solution of activation more than 1 hour down, uses CaHPO
4Transfer H7.2, last dextran G-25 chromatography.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN 92112188 CN1074709A (en) | 1992-11-26 | 1992-11-26 | Method for preparing thrombin |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN 92112188 CN1074709A (en) | 1992-11-26 | 1992-11-26 | Method for preparing thrombin |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN1074709A true CN1074709A (en) | 1993-07-28 |
Family
ID=4945802
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN 92112188 Pending CN1074709A (en) | 1992-11-26 | 1992-11-26 | Method for preparing thrombin |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN1074709A (en) |
Cited By (17)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US5585007A (en) * | 1994-12-07 | 1996-12-17 | Plasmaseal Corporation | Plasma concentrate and tissue sealant methods and apparatuses for making concentrated plasma and/or tissue sealant |
| CN1063789C (en) * | 1996-04-06 | 2001-03-28 | 济南军区生物制品药物研究所 | method for extracting and separating thrombin of beef |
| CN1294257C (en) * | 2004-09-15 | 2007-01-10 | 黄耀江 | Process for preparing thrombase |
| US8801586B2 (en) | 2008-02-29 | 2014-08-12 | Biomet Biologics, Llc | System and process for separating a material |
| CN105821025A (en) * | 2016-04-06 | 2016-08-03 | 浙江丰安生物制药有限公司 | Extraction method of thrombin |
| US9642956B2 (en) | 2012-08-27 | 2017-05-09 | Biomet Biologics, Llc | Apparatus and method for separating and concentrating fluids containing multiple components |
| US9649579B2 (en) | 2007-04-12 | 2017-05-16 | Hanuman Llc | Buoy suspension fractionation system |
| US9701728B2 (en) | 2008-02-27 | 2017-07-11 | Biomet Biologics, Llc | Methods and compositions for delivering interleukin-1 receptor antagonist |
| US9713810B2 (en) | 2015-03-30 | 2017-07-25 | Biomet Biologics, Llc | Cell washing plunger using centrifugal force |
| US9757721B2 (en) | 2015-05-11 | 2017-09-12 | Biomet Biologics, Llc | Cell washing plunger using centrifugal force |
| US9897589B2 (en) | 2002-05-24 | 2018-02-20 | Biomet Biologics, Llc | Apparatus and method for separating and concentrating fluids containing multiple components |
| US9895418B2 (en) | 2013-03-15 | 2018-02-20 | Biomet Biologics, Llc | Treatment of peripheral vascular disease using protein solutions |
| US9950035B2 (en) | 2013-03-15 | 2018-04-24 | Biomet Biologics, Llc | Methods and non-immunogenic compositions for treating inflammatory disorders |
| US10143725B2 (en) | 2013-03-15 | 2018-12-04 | Biomet Biologics, Llc | Treatment of pain using protein solutions |
| US10183042B2 (en) | 2002-05-24 | 2019-01-22 | Biomet Manufacturing, Llc | Apparatus and method for separating and concentrating fluids containing multiple components |
| US10208095B2 (en) | 2013-03-15 | 2019-02-19 | Biomet Manufacturing, Llc | Methods for making cytokine compositions from tissues using non-centrifugal methods |
| US10576130B2 (en) | 2013-03-15 | 2020-03-03 | Biomet Manufacturing, Llc | Treatment of collagen defects using protein solutions |
-
1992
- 1992-11-26 CN CN 92112188 patent/CN1074709A/en active Pending
Cited By (25)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US5585007A (en) * | 1994-12-07 | 1996-12-17 | Plasmaseal Corporation | Plasma concentrate and tissue sealant methods and apparatuses for making concentrated plasma and/or tissue sealant |
| US6214338B1 (en) | 1994-12-07 | 2001-04-10 | Plasmaseal Llc | Plasma concentrate and method of processing blood for same |
| CN1063789C (en) * | 1996-04-06 | 2001-03-28 | 济南军区生物制品药物研究所 | method for extracting and separating thrombin of beef |
| US10183042B2 (en) | 2002-05-24 | 2019-01-22 | Biomet Manufacturing, Llc | Apparatus and method for separating and concentrating fluids containing multiple components |
| US9897589B2 (en) | 2002-05-24 | 2018-02-20 | Biomet Biologics, Llc | Apparatus and method for separating and concentrating fluids containing multiple components |
| US10393728B2 (en) | 2002-05-24 | 2019-08-27 | Biomet Biologics, Llc | Apparatus and method for separating and concentrating fluids containing multiple components |
| CN1294257C (en) * | 2004-09-15 | 2007-01-10 | 黄耀江 | Process for preparing thrombase |
| US9649579B2 (en) | 2007-04-12 | 2017-05-16 | Hanuman Llc | Buoy suspension fractionation system |
| US9701728B2 (en) | 2008-02-27 | 2017-07-11 | Biomet Biologics, Llc | Methods and compositions for delivering interleukin-1 receptor antagonist |
| US10400017B2 (en) | 2008-02-27 | 2019-09-03 | Biomet Biologics, Llc | Methods and compositions for delivering interleukin-1 receptor antagonist |
| US11725031B2 (en) | 2008-02-27 | 2023-08-15 | Biomet Manufacturing, Llc | Methods and compositions for delivering interleukin-1 receptor antagonist |
| US9719063B2 (en) | 2008-02-29 | 2017-08-01 | Biomet Biologics, Llc | System and process for separating a material |
| US8801586B2 (en) | 2008-02-29 | 2014-08-12 | Biomet Biologics, Llc | System and process for separating a material |
| US9642956B2 (en) | 2012-08-27 | 2017-05-09 | Biomet Biologics, Llc | Apparatus and method for separating and concentrating fluids containing multiple components |
| US9950035B2 (en) | 2013-03-15 | 2018-04-24 | Biomet Biologics, Llc | Methods and non-immunogenic compositions for treating inflammatory disorders |
| US10143725B2 (en) | 2013-03-15 | 2018-12-04 | Biomet Biologics, Llc | Treatment of pain using protein solutions |
| US9895418B2 (en) | 2013-03-15 | 2018-02-20 | Biomet Biologics, Llc | Treatment of peripheral vascular disease using protein solutions |
| US10208095B2 (en) | 2013-03-15 | 2019-02-19 | Biomet Manufacturing, Llc | Methods for making cytokine compositions from tissues using non-centrifugal methods |
| US10441634B2 (en) | 2013-03-15 | 2019-10-15 | Biomet Biologics, Llc | Treatment of peripheral vascular disease using protein solutions |
| US10576130B2 (en) | 2013-03-15 | 2020-03-03 | Biomet Manufacturing, Llc | Treatment of collagen defects using protein solutions |
| US11957733B2 (en) | 2013-03-15 | 2024-04-16 | Biomet Manufacturing, Llc | Treatment of collagen defects using protein solutions |
| US9713810B2 (en) | 2015-03-30 | 2017-07-25 | Biomet Biologics, Llc | Cell washing plunger using centrifugal force |
| US9757721B2 (en) | 2015-05-11 | 2017-09-12 | Biomet Biologics, Llc | Cell washing plunger using centrifugal force |
| CN105821025B (en) * | 2016-04-06 | 2019-06-28 | 浙江丰安生物制药有限公司 | A kind of extracting method of fibrin ferment |
| CN105821025A (en) * | 2016-04-06 | 2016-08-03 | 浙江丰安生物制药有限公司 | Extraction method of thrombin |
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