CS256710B1 - Method of albumin extraction from cohne fraction 4 - Google Patents
Method of albumin extraction from cohne fraction 4 Download PDFInfo
- Publication number
- CS256710B1 CS256710B1 CS855203A CS520385A CS256710B1 CS 256710 B1 CS256710 B1 CS 256710B1 CS 855203 A CS855203 A CS 855203A CS 520385 A CS520385 A CS 520385A CS 256710 B1 CS256710 B1 CS 256710B1
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- CS
- Czechoslovakia
- Prior art keywords
- albumin
- fraction
- cohne
- cohn fraction
- solution
- Prior art date
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- 102000009027 Albumins Human genes 0.000 title claims description 27
- 108010088751 Albumins Proteins 0.000 title claims description 27
- 238000000034 method Methods 0.000 title claims description 13
- 238000000605 extraction Methods 0.000 title 1
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 claims description 13
- 108010032608 Cohn fraction IV Proteins 0.000 claims description 9
- 108010017384 Blood Proteins Proteins 0.000 claims description 5
- 102000004506 Blood Proteins Human genes 0.000 claims description 5
- DZTHIGRZJZPRDV-UHFFFAOYSA-N N-acetyltryptophan Chemical compound C1=CC=C2C(CC(NC(=O)C)C(O)=O)=CNC2=C1 DZTHIGRZJZPRDV-UHFFFAOYSA-N 0.000 claims description 4
- 239000002244 precipitate Substances 0.000 claims description 3
- 102000004169 proteins and genes Human genes 0.000 claims description 2
- 108090000623 proteins and genes Proteins 0.000 claims description 2
- 239000000047 product Substances 0.000 description 4
- 238000004925 denaturation Methods 0.000 description 3
- 230000036425 denaturation Effects 0.000 description 3
- 238000005194 fractionation Methods 0.000 description 3
- 238000011084 recovery Methods 0.000 description 3
- 238000005119 centrifugation Methods 0.000 description 2
- 238000004108 freeze drying Methods 0.000 description 2
- 238000002955 isolation Methods 0.000 description 2
- 238000001556 precipitation Methods 0.000 description 2
- 239000006228 supernatant Substances 0.000 description 2
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 2
- 108010054147 Hemoglobins Proteins 0.000 description 1
- 102000001554 Hemoglobins Human genes 0.000 description 1
- DGAQECJNVWCQMB-PUAWFVPOSA-M Ilexoside XXIX Chemical compound C[C@@H]1CC[C@@]2(CC[C@@]3(C(=CC[C@H]4[C@]3(CC[C@@H]5[C@@]4(CC[C@@H](C5(C)C)OS(=O)(=O)[O-])C)C)[C@@H]2[C@]1(C)O)C)C(=O)O[C@H]6[C@@H]([C@H]([C@@H]([C@H](O6)CO)O)O)O.[Na+] DGAQECJNVWCQMB-PUAWFVPOSA-M 0.000 description 1
- 238000006243 chemical reaction Methods 0.000 description 1
- 239000003153 chemical reaction reagent Substances 0.000 description 1
- -1 citrate ions Chemical class 0.000 description 1
- 230000006378 damage Effects 0.000 description 1
- 230000007423 decrease Effects 0.000 description 1
- 238000011156 evaluation Methods 0.000 description 1
- 230000036512 infertility Effects 0.000 description 1
- 238000007689 inspection Methods 0.000 description 1
- 238000002360 preparation method Methods 0.000 description 1
- 230000000750 progressive effect Effects 0.000 description 1
- 238000000746 purification Methods 0.000 description 1
- 230000001698 pyrogenic effect Effects 0.000 description 1
- 229910052708 sodium Inorganic materials 0.000 description 1
- 239000011734 sodium Substances 0.000 description 1
- 230000006641 stabilisation Effects 0.000 description 1
- 238000011105 stabilization Methods 0.000 description 1
- 239000003381 stabilizer Substances 0.000 description 1
Landscapes
- Peptides Or Proteins (AREA)
Description
Vynález sa týká sposobu získavania albuminu z Cohnovej frakcie IV, vznikajúcej pri etanolovej frakcionácii ludskej plazmy, a to tým spůsobom, že plazmatické bielkoviny doprevádzajúce albumin sa oddelujú pri vhodnom pH, teplote a koncentráci etanolu vo formě precipitátu a albumin ostává v roztoku.The invention relates to a process for recovering albumin from Cohn Fraction IV resulting from ethanol fractionation of human plasma, in that the plasma proteins accompanying albumin are separated at a suitable pH, temperature and ethanol concentration in the form of a precipitate and the albumin remains in solution.
Jedna z možnosti zvýšenia výťažnosti albuminu pri frakcionácii ludskej plazmy je cesta zavádzania nových progresívnych technologických postupov, V poeledných rokoch na dosiahnutie maximálnej výtažnosti albuminu je evidentný trend, zavádzania technologických stupňov a operách znižuje, čím aj straty na bielkovine albuminu sú nižšie. Medzi takéto frakcionačné postupy možno zaradiť aj tepelno-denaturačné metódy izolácie albuminu (Hoch H., Chanutin A.: Arch. of biochem. and biophysic, 51, 27, 1954; Lundblad J. I.: Vox sang., 5, 122, 1960; Gabr Y., Soliman H. R.: Acta biol. et med. ger., 27, 341, 1971; Schneider W., Lefevre H., Friedler H., Mc Carty L. I.: Blut, 30, 121, rok 1975; Wye E. Y. and Kim Μ. K.: Vof Sang., 32,182 — 184,1977).One possibility of increasing albumin recovery in human plasma fractionation is the way to introduce new progressive technological processes. In recent years to achieve maximum albumin recovery, there is an evident trend, the introduction of technological steps and operations decreases, and even losses in albumin protein are lower. Such fractionation techniques include thermal denaturation methods for albumin isolation (Hoch H., Chanutin A .: Arch. Of Biochem. And Biophysic, 51, 27, 1954; Lundblad JI: Vox sang., 5, 122, 1960; Gabr). Y., Soliman HR: Acta Biol et Med., 27, 341, 1971, Schneider W., Lefevre H., Friedler H., Mc Carty LI: Blut, 30, 121, 1975; Wye EY and Kim K K .: Vof Sang., 32, 182-184, 1977).
Negatívom citovaných technologických postupov je skutočnosť, že tieto pracujú pri vysokých teplotách (+70°Cj, při ktorých napriek stabilizácii může albumin za určitých podmienok vykazovat nežiadúce inreverzibilné změny.A disadvantage of the cited technological processes is that they operate at high temperatures (+ 70 ° Cj), at which, despite stabilization, albumin may exhibit undesirable inversible changes under certain conditions.
Na základe uvedeného boli vyvíjané snahy čiastočné znížiť denaturačné teploty a tým predchádzať eventuálnej deštrukcii izolovaného albuminu (Libich M.: PV 74-57-79, Foukal Z.: PV 05-36-81, Bulík J.: AO 206 294).Accordingly, efforts have been made to partially reduce denaturation temperatures and thereby prevent eventual destruction of isolated albumin (Libich M .: PV 74-57-79, Foukal Z .: PV 05-36-81, Bulík J .: AO 206 294).
Napriek týmto pozitívam vo váčšine prípadov používané denaturačné teploty sú stále hraničně, pričom technologické postupy sú poměrně členité, pozostávajúce z viacerých technologických stupňov.Despite these positives, in most cases the denaturation temperatures used are still marginal, and the technological processes are quite rugged, consisting of several technological stages.
Novost navrhovaného technologického postupu získavania albuminu z Cohnovej frakcie IV je, že tento vdaka vhodnej koncentrácii stabilizátorov, organického činidla a hodnoty pH dovoluje podstatné znížiť reakčnú teplotu a skrátiť technologický postup pri garancii nativity izolovanej bielkoviny.The novelty of the proposed process for recovering albumin from Cohn Fraction IV is that due to the appropriate concentration of stabilizers, organic reagent and pH, it allows to substantially reduce the reaction temperature and shorten the process of guaranteeing the nativeity of the isolated protein.
Podstata získavania albuminu z Cohnovej frakcie IV spočívá v tom, že 2- až 4 %-ný roztok Cohnovej frakcie IV sa stabilizujeThe essence of recovering albumin from Cohn Fraction IV is that the 2-4% Cohn Fraction IV solution is stabilized
0,03 M Na-kaprylánom a 0,01 M acetyl-DL-tryptofánom a pri pH 5,0 až 5,5, koncentrácii etanolu 5,0 až 9,0 objemových % a teplote +40 až 45 °C po dobu 0,5: až 12 hodin sa vyprecipitujú albumin doprevádzajúce plazmatické bielkoviny a albumin ostává v roztoku. Koncentrácia a purifikácia roztoku albuminu sa dalej prevádza známými technologickými operáciami.0.03 M Na-caprylan and 0.01 M acetyl-DL-tryptophan and at pH 5.0-5.5, ethanol concentration 5.0-9.0% v / v and temperature +40-45 ° C for 0 5: 12 to 12 hours, the albumin accompanying the plasma proteins precipitates and the albumin remains in solution. The concentration and purification of the albumin solution is further carried out by known technological operations.
Výhoda popisaného technologického postupu získavania albuminu z Cohnovej frakcie IV oproti doteraz používanému spočívá v tom, že technologický proces sa skracuje na jeden technologický stupeň. Uvedeným izolačným postupom je garantovaná aj kvalita, najmá však nativita získaného albuminu s evidentnou úsporou energií.The advantage of the disclosed process for recovering albumin from Cohn Fraction IV over the prior art is that the process is shortened to one process stage. This isolation procedure also guarantees quality, but in particular the nativity of the albumin obtained with evident energy savings.
Příklad 1Example 1
300 g Cohnovej frakcie IV sa suspenduje v 35 000 ml apyrogénnej vody. K takto připravenému roztoku sa přidá 154 g Na-kaprylátu a 86,1 g acetyl-DL-tryptofánu. Hodnota pH sa upraví na 5,1, koncentrácia etanolu na 7 objemových % a teplota na +40 stupňov Celzia. Precipitácia plazmatických bielkovín doprevádzajúcich albumin prebieha po dobu 10 hodin. Vyprecipitované plazmatické bielkoviny sa separujú centrifugáciou a supernatant obsahujúci albumin sa dalej purifikuje a koncentruje lyofilizáciou. Hmotnost lyof ilizovaného produktu je 1120 gramu elektroforetická čistota 96,1 %. Příklad 2300 g of Cohn Fraction IV are suspended in 35,000 ml of pyrogen-free water. To this solution was added 154 g of Na-caprylate and 86.1 g of acetyl-DL-tryptophan. The pH is adjusted to 5.1, the ethanol concentration to 7% by volume and the temperature to +40 degrees Celsius. The precipitation of albumin accompanying albumin takes place for 10 hours. The precipitated plasma proteins are separated by centrifugation and the albumin-containing supernatant is further purified and concentrated by lyophilization. The weight of the lyophilized product is 1120 grams electrophoretic purity 96.1%. Example 2
650 g Cohnovej frakcie IV sa supenduje v 17 500 ml apyrogénnej vody. K takto připravenému roztoku sa potom přidá 77,0 g Na-kaprylánu a 43,0 g acetyl-DL-tryptofánu. Hodnota pH sa upraví na 5,2, koncentrácia etanolu 9 objemových % a teplota na +45 stupňov Celzia. Precipitácia plazmatických bielkovín doprevádzajúcich albumin prebieha po dobu 12 hodin. Vyprecipitované plazmatické bielkoviny sa separujú centrifugáciou a supernatant obsahujúci albumin sa dalej purifikuje a koncentruje lyofilizáciou. Hmotnosť lyofilizovaného produktu je 490 g, elektroforetická čistota 96,2 %.650 g of Cohn Fraction IV are suspended in 17,500 ml of pyrogen-free water. 77.0 g of Na-caprylane and 43.0 g of acetyl-DL-tryptophan are then added to the solution. The pH is adjusted to 5.2, the ethanol concentration is 9% by volume and the temperature to +45 degrees Celsius. The precipitation of albumin accompanying albumin takes place for 12 hours. The precipitated plasma proteins are separated by centrifugation and the albumin-containing supernatant is further purified and concentrated by lyophilization. The weight of the lyophilized product is 490 g, electrophoretic purity 96.2%.
Hodnotenie kvality přípravkuEvaluation of product quality
Požadované skúšky Kritéria Výsledky kontrolyRequired tests Criteria Results of inspection
PNY 31-07-79 přípravkuPNY 31-07-79 preparation
Dosahovaná výťažnosť albuminu sa pohybuje pódia jeho obsahu v Cohnovej frakciiThe albumin recovery achieved is based on its content in the Cohn fraction
IV v rozmedzí 100 až 160 g lyofilizovaného produktu na 1 000 g frakcie IV.IV in the range of 100 to 160 g of lyophilized product per 1000 g of Fraction IV.
Claims (1)
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CS855203A CS256710B1 (en) | 1985-07-12 | 1985-07-12 | Method of albumin extraction from cohne fraction 4 |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CS855203A CS256710B1 (en) | 1985-07-12 | 1985-07-12 | Method of albumin extraction from cohne fraction 4 |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CS520385A1 CS520385A1 (en) | 1987-09-17 |
| CS256710B1 true CS256710B1 (en) | 1988-04-15 |
Family
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Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CS855203A CS256710B1 (en) | 1985-07-12 | 1985-07-12 | Method of albumin extraction from cohne fraction 4 |
Country Status (1)
| Country | Link |
|---|---|
| CS (1) | CS256710B1 (en) |
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1985
- 1985-07-12 CS CS855203A patent/CS256710B1/en unknown
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| Publication number | Publication date |
|---|---|
| CS520385A1 (en) | 1987-09-17 |
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