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RU2014123868A - METHOD FOR PRODUCING THROMBIN CONCENTRATE - Google Patents

METHOD FOR PRODUCING THROMBIN CONCENTRATE Download PDF

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Publication number
RU2014123868A
RU2014123868A RU2014123868/15A RU2014123868A RU2014123868A RU 2014123868 A RU2014123868 A RU 2014123868A RU 2014123868/15 A RU2014123868/15 A RU 2014123868/15A RU 2014123868 A RU2014123868 A RU 2014123868A RU 2014123868 A RU2014123868 A RU 2014123868A
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RU
Russia
Prior art keywords
prothrombin
cacl
thrombin
solution
isolation
Prior art date
Application number
RU2014123868/15A
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Russian (ru)
Other versions
RU2583931C2 (en
Inventor
Арон Леонидович Берковский
Александр Владимирович Суворов
Евгений Михайлович Голубев
Елена Владимировна Сергеева
Татьяна Леонидовна Дереза
Оксана Георгиевна Кутюрова
Вячеслав Викторович Хурдин
Анастасия Викторовна Слободян
Original Assignee
Федеральное государственное бюджетное учреждение Гематологический научный центр Министерства здравоохранения РФ
Priority date (The priority date 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 date listed.)
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Application filed by Федеральное государственное бюджетное учреждение Гематологический научный центр Министерства здравоохранения РФ filed Critical Федеральное государственное бюджетное учреждение Гематологический научный центр Министерства здравоохранения РФ
Priority to RU2014123868/15A priority Critical patent/RU2583931C2/en
Publication of RU2014123868A publication Critical patent/RU2014123868A/en
Application granted granted Critical
Publication of RU2583931C2 publication Critical patent/RU2583931C2/en

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    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K35/00—Medicinal preparations containing materials or reaction products thereof with undetermined constitution
    • A61K35/12—Materials from mammals; Compositions comprising non-specified tissues or cells; Compositions comprising non-embryonic stem cells; Genetically modified cells
    • A61K35/14—Blood; Artificial blood
    • A61K35/16—Blood plasma; Blood serum
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K1/00—General methods for the preparation of peptides, i.e. processes for the organic chemical preparation of peptides or proteins of any length
    • C07K1/14—Extraction; Separation; Purification
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K14/00—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
    • C07K14/435—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans
    • C07K14/745—Blood coagulation or fibrinolysis factors
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N9/00—Enzymes; Proenzymes; Compositions thereof; Processes for preparing, activating, inhibiting, separating or purifying enzymes
    • C12N9/14—Hydrolases (3)
    • C12N9/48—Hydrolases (3) acting on peptide bonds (3.4)
    • C12N9/50—Proteinases, e.g. Endopeptidases (3.4.21-3.4.25)
    • C12N9/64—Proteinases, e.g. Endopeptidases (3.4.21-3.4.25) derived from animal tissue
    • C12N9/6421—Proteinases, e.g. Endopeptidases (3.4.21-3.4.25) derived from animal tissue from mammals
    • C12N9/6424—Serine endopeptidases (3.4.21)
    • C12N9/6429—Thrombin (3.4.21.5)

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  • Health & Medical Sciences (AREA)
  • Chemical & Material Sciences (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Organic Chemistry (AREA)
  • General Health & Medical Sciences (AREA)
  • Genetics & Genomics (AREA)
  • Medicinal Chemistry (AREA)
  • Zoology (AREA)
  • Engineering & Computer Science (AREA)
  • Biochemistry (AREA)
  • Hematology (AREA)
  • Molecular Biology (AREA)
  • Biomedical Technology (AREA)
  • Biotechnology (AREA)
  • Cell Biology (AREA)
  • Proteomics, Peptides & Aminoacids (AREA)
  • Biophysics (AREA)
  • Wood Science & Technology (AREA)
  • Bioinformatics & Cheminformatics (AREA)
  • Public Health (AREA)
  • Immunology (AREA)
  • Pharmacology & Pharmacy (AREA)
  • Epidemiology (AREA)
  • Animal Behavior & Ethology (AREA)
  • Virology (AREA)
  • Veterinary Medicine (AREA)
  • Analytical Chemistry (AREA)
  • Microbiology (AREA)
  • Developmental Biology & Embryology (AREA)
  • General Engineering & Computer Science (AREA)
  • Toxicology (AREA)
  • Gastroenterology & Hepatology (AREA)
  • Medicines That Contain Protein Lipid Enzymes And Other Medicines (AREA)
  • Peptides Or Proteins (AREA)
  • Enzymes And Modification Thereof (AREA)

Abstract

Способ выделения очищенного концентрата тромбина, избавленного от вирусов, характеризующегося большим выходом, чистотой и высокой стабильностью, заключающийся в криофракционировании исходного сырья, представленного свежезамороженной плазмой донорской крови человека, выделении протромбинового комплекса на анионообменном сорбенте DEAE Sepharose Fast Flow, активации протромбина раствором CaClв составе протромбинового комплекса с получением тромбина, вирусной инактивации сольвент - детергентным методом и очистке полученного тромбина на катионообменном сорбенте Fractogel EMD-SO, отличающийся тем, что количество раствора CaCl, выступающего в качестве активатора протромбина, рассчитывается с учетом взаимодействия CaClс цитратом натрия, который входит в состав буфера, используемого при выделении протромбинового комплекса на анионобменном сорбенте, так, чтобы концентрация свободных ионов Сав растворе на стадии активации составляла от 2 до 20 ммоль/л.A method for isolating a purified thrombin concentrate, freed from viruses, characterized by a high yield, purity, and high stability, which consists in cryofractionation of the feedstock represented by freshly frozen human blood donated plasma, isolation of the prothrombin complex on the DEAE Sepharose Fast Flow anion-exchange sorbent, prothrombin activation with CaCl solution in the prothrombin complex with obtaining thrombin, viral inactivation of the solvent - detergent method and purification of the obtained thrombin on cation Fractogel EMD-SO belt sorbent, characterized in that the amount of CaCl solution acting as a prothrombin activator is calculated taking into account the interaction of CaCl with sodium citrate, which is part of the buffer used in the isolation of the prothrombin complex on the anion exchange sorbent, so that the concentration of free ions Cav solution at the stage of activation ranged from 2 to 20 mmol / L.

Claims (1)

Способ выделения очищенного концентрата тромбина, избавленного от вирусов, характеризующегося большим выходом, чистотой и высокой стабильностью, заключающийся в криофракционировании исходного сырья, представленного свежезамороженной плазмой донорской крови человека, выделении протромбинового комплекса на анионообменном сорбенте DEAE Sepharose Fast Flow, активации протромбина раствором CaCl2 в составе протромбинового комплекса с получением тромбина, вирусной инактивации сольвент - детергентным методом и очистке полученного тромбина на катионообменном сорбенте Fractogel EMD-SO3, отличающийся тем, что количество раствора CaCl2, выступающего в качестве активатора протромбина, рассчитывается с учетом взаимодействия CaCl2 с цитратом натрия, который входит в состав буфера, используемого при выделении протромбинового комплекса на анионобменном сорбенте, так, чтобы концентрация свободных ионов Са2+ в растворе на стадии активации составляла от 2 до 20 ммоль/л. A method for isolating a purified thrombin concentrate, freed from viruses, characterized by a high yield, purity and high stability, which consists in cryofractionation of the feedstock represented by freshly frozen human donor blood plasma, isolation of the prothrombin complex on an anion exchange sorbent DEAE Sepharose Fast Flow, prothrombin activation with CaCl 2 solution in the composition prothrombin complex to obtain thrombin, viral solvent inactivation by the detergent method and purification of the obtained thrombin to cation Fractogel EMD-SO 3 exchange sorbent, characterized in that the amount of CaCl 2 solution acting as a prothrombin activator is calculated taking into account the interaction of CaCl 2 with sodium citrate, which is part of the buffer used in the isolation of the prothrombin complex on an anion exchange sorbent, so that the concentration of free Ca 2+ ions in the solution at the activation stage is from 2 to 20 mmol / L.
RU2014123868/15A 2014-06-11 2014-06-11 Method of producing thrombin concentrate RU2583931C2 (en)

Priority Applications (1)

Application Number Priority Date Filing Date Title
RU2014123868/15A RU2583931C2 (en) 2014-06-11 2014-06-11 Method of producing thrombin concentrate

Applications Claiming Priority (1)

Application Number Priority Date Filing Date Title
RU2014123868/15A RU2583931C2 (en) 2014-06-11 2014-06-11 Method of producing thrombin concentrate

Publications (2)

Publication Number Publication Date
RU2014123868A true RU2014123868A (en) 2015-12-20
RU2583931C2 RU2583931C2 (en) 2016-05-10

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RU2014123868/15A RU2583931C2 (en) 2014-06-11 2014-06-11 Method of producing thrombin concentrate

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Cited By (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN118460516A (en) * 2024-05-28 2024-08-09 成都协和生物技术有限责任公司 An activated prothrombin complex aPCC and its preparation method and application

Family Cites Families (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US5981254A (en) * 1997-10-30 1999-11-09 Haemacure Corporation Process for producing thrombin from plasma
DE10012732A1 (en) * 2000-03-18 2001-09-20 Aventis Behring Gmbh Thrombin composition, for use as hemostatic or as a component of fibrin glues, comprises non-covalently bonded inhibitor for stabilization
RU2457248C2 (en) * 2010-06-09 2012-07-27 Общество с ограниченной ответственностью фирма "Технология-Стандарт" Method for commercial production of thrombin

Cited By (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN118460516A (en) * 2024-05-28 2024-08-09 成都协和生物技术有限责任公司 An activated prothrombin complex aPCC and its preparation method and application

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RH4A Copy of patent granted that was duplicated for the russian federation

Effective date: 20200901