CA2476177A1 - Purification of red blood cells by separation and diafiltration - Google Patents

Purification of red blood cells by separation and diafiltration Download PDF

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Publication number
CA2476177A1
CA2476177A1 CA002476177A CA2476177A CA2476177A1 CA 2476177 A1 CA2476177 A1 CA 2476177A1 CA 002476177 A CA002476177 A CA 002476177A CA 2476177 A CA2476177 A CA 2476177A CA 2476177 A1 CA2476177 A1 CA 2476177A1
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Prior art keywords
red blood
whole blood
blood cells
cells
purified
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CA002476177A
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French (fr)
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CA2476177C (en
Inventor
Robert A. Houtchens
Maria S. Gawryl
William R. Light
Javed Baqai
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HEMOGLOBIN OXYGEN THERAPEUTICS LLC
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Individual
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Expired - Fee Related legal-status Critical Current

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    • 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
    • C12N5/00—Undifferentiated human, animal or plant cells, e.g. cell lines; Tissues; Cultivation or maintenance thereof; Culture media therefor
    • C12N5/06—Animal cells or tissues; Human cells or tissues
    • C12N5/0602—Vertebrate cells
    • C12N5/0634—Cells from the blood or the immune system
    • C12N5/0641—Erythrocytes
    • 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
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K38/00—Medicinal preparations containing peptides
    • A61K38/16—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
    • A61K38/41—Porphyrin- or corrin-ring-containing peptides
    • A61K38/42—Haemoglobins; Myoglobins
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61M—DEVICES FOR INTRODUCING MEDIA INTO, OR ONTO, THE BODY; DEVICES FOR TRANSDUCING BODY MEDIA OR FOR TAKING MEDIA FROM THE BODY; DEVICES FOR PRODUCING OR ENDING SLEEP OR STUPOR
    • A61M1/00—Suction or pumping devices for medical purposes; Devices for carrying-off, for treatment of, or for carrying-over, body-liquids; Drainage systems
    • A61M1/36—Other treatment of blood in a by-pass of the natural circulatory system, e.g. temperature adaptation, irradiation ; Extra-corporeal blood circuits
    • A61M1/3692—Washing or rinsing blood or blood constituents
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61M—DEVICES FOR INTRODUCING MEDIA INTO, OR ONTO, THE BODY; DEVICES FOR TRANSDUCING BODY MEDIA OR FOR TAKING MEDIA FROM THE BODY; DEVICES FOR PRODUCING OR ENDING SLEEP OR STUPOR
    • A61M1/00—Suction or pumping devices for medical purposes; Devices for carrying-off, for treatment of, or for carrying-over, body-liquids; Drainage systems
    • A61M1/36—Other treatment of blood in a by-pass of the natural circulatory system, e.g. temperature adaptation, irradiation ; Extra-corporeal blood circuits
    • A61M1/3693—Other treatment of blood in a by-pass of the natural circulatory system, e.g. temperature adaptation, irradiation ; Extra-corporeal blood circuits using separation based on different densities of components, e.g. centrifuging
    • A61M1/3696—Other treatment of blood in a by-pass of the natural circulatory system, e.g. temperature adaptation, irradiation ; Extra-corporeal blood circuits using separation based on different densities of components, e.g. centrifuging with means for adding or withdrawing liquid substances during the centrifugation, e.g. continuous centrifugation
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P7/00—Drugs for disorders of the blood or the extracellular fluid
    • A61P7/08—Plasma substitutes; Perfusion solutions; Dialytics or haemodialytics; Drugs for electrolytic or acid-base disorders, e.g. hypovolemic shock
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61M—DEVICES FOR INTRODUCING MEDIA INTO, OR ONTO, THE BODY; DEVICES FOR TRANSDUCING BODY MEDIA OR FOR TAKING MEDIA FROM THE BODY; DEVICES FOR PRODUCING OR ENDING SLEEP OR STUPOR
    • A61M1/00—Suction or pumping devices for medical purposes; Devices for carrying-off, for treatment of, or for carrying-over, body-liquids; Drainage systems
    • A61M1/36—Other treatment of blood in a by-pass of the natural circulatory system, e.g. temperature adaptation, irradiation ; Extra-corporeal blood circuits
    • A61M1/3693—Other treatment of blood in a by-pass of the natural circulatory system, e.g. temperature adaptation, irradiation ; Extra-corporeal blood circuits using separation based on different densities of components, e.g. centrifuging
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61M—DEVICES FOR INTRODUCING MEDIA INTO, OR ONTO, THE BODY; DEVICES FOR TRANSDUCING BODY MEDIA OR FOR TAKING MEDIA FROM THE BODY; DEVICES FOR PRODUCING OR ENDING SLEEP OR STUPOR
    • A61M2202/00—Special media to be introduced, removed or treated
    • A61M2202/04—Liquids
    • A61M2202/0413—Blood
    • A61M2202/0429—Red blood cells; Erythrocytes

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  • Health & Medical Sciences (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Engineering & Computer Science (AREA)
  • Biomedical Technology (AREA)
  • General Health & Medical Sciences (AREA)
  • Hematology (AREA)
  • Vascular Medicine (AREA)
  • Heart & Thoracic Surgery (AREA)
  • Veterinary Medicine (AREA)
  • Chemical & Material Sciences (AREA)
  • Animal Behavior & Ethology (AREA)
  • Public Health (AREA)
  • Bioinformatics & Cheminformatics (AREA)
  • Zoology (AREA)
  • Organic Chemistry (AREA)
  • Biotechnology (AREA)
  • Immunology (AREA)
  • Anesthesiology (AREA)
  • Cell Biology (AREA)
  • Genetics & Genomics (AREA)
  • Wood Science & Technology (AREA)
  • Cardiology (AREA)
  • Pharmacology & Pharmacy (AREA)
  • Medicinal Chemistry (AREA)
  • Epidemiology (AREA)
  • Microbiology (AREA)
  • Biochemistry (AREA)
  • General Engineering & Computer Science (AREA)
  • Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
  • Proteomics, Peptides & Aminoacids (AREA)
  • Developmental Biology & Embryology (AREA)
  • Virology (AREA)
  • Gastroenterology & Hepatology (AREA)
  • General Chemical & Material Sciences (AREA)
  • Chemical Kinetics & Catalysis (AREA)
  • Diabetes (AREA)
  • Medicines Containing Material From Animals Or Micro-Organisms (AREA)
  • Micro-Organisms Or Cultivation Processes Thereof (AREA)
  • External Artificial Organs (AREA)

Abstract

Red blood cells are purified by separating whole blood, such as by centrifugation, to form a red blood cell fraction and a liquid fraction. The whole blood can be defibrinated or treated to prevent coagulation prior to separation. Preferably, the whole blood is bovine blood. The red blood cell fraction is then diafiltered to purify the red blood cells. The purified red blood cells can then be lysed to form a lysate of purified red blood cells.
The purified red blood cells and the lysate of purified red blood cells are suitable for use in producing hemoglobin blood substitute.

Claims (27)

1. A method of purifying red blood cells, comprising the steps;
a) separating whole blood, whereby a red blood cell fraction and a liquid fraction are formed; and b) diafiltering the red blood cell fraction to thereby form purified red blood cells.
2. The method of Claim 1, wherein the whole blood is separated by sedimentation of red blood cells in the whole blood.
3. The method of Claim 2, wherein the sedimentation of red blood cells is obtained by centrifuging the whole blood.
4. The method of Claim 3, wherein the centrifugation of the whole blood causes the red blood cell fraction to consist essentially of red blood cells.
5. The method of Claim 1, wherein the whole blood is fractionated by exposing the whole blood to a G-force in a range of between about 10 x G and about 12,000 x G.
6. The method of Claim 1, wherein the liquid fraction is removed from the from the red blood cell fraction by decanting after step a).
7. The method of Claim 1, wherein the liquid fraction is removed from the red blood cell fraction simultaneously with separation of the liquid fraction and the red blood cell fraction.
8. The method of Claim 1, wherein the whole blood is defibrinated.
9. The method of Claim 8, wherein the whole blood is defibrinated mechanically.
10. The method of Claim 1, wherein the whole blood is treated with an anticoagulant.
11. The method of Claim 10, wherein the anticoagulant is selected from the group consisting of: sodium citrate, heparin, ethylenediaminetetraacetic acid (EDTA) and sodium oxylate.
12. The method of Claim 11, wherein the anticoagulant is sodium citrate.
13. The method of Claim 11, wherein the anticoagulant is heparin.
14. The method of Claim 1, further including the step of lysing the purified red blood cells.
15. The method of Claim 14, wherein the purified red blood cells are lysed mechanically.
16. The method of Claim 14, wherein the purified red blood cells are lysed osmotically.
17. The method of Claim 1, wherein the liquid fraction includes most of red cells of the whole blood.
18. The method of Claim 17, wherein the red blood cell fraction includes most of the white cells and platelets of the whole blood.
19. The method of Claim 1, wherein the red blood cell fraction is diafiltered with a membrane having a permeability in a range of between about 0.1 µm and about 5 µm.
20. The method of Claim 1, wherein the whole blood is bovine whole blood.
21. A method of forming a lysate of purified red blood cells for use in a hemoglobin blood substitute, comprising the steps;
a) separating whole blood, whereby a red blood cell fraction and a liquid fraction are formed;
b) diafiltering the red blood cell fraction to thereby foam purified red blood cells;
and c) lysing the purified red blood cells, thereby forming the lysate of purified red blood cells.
22. The method of Claim 21, wherein the whole blood is mechanically defibrinated.
23. The method of Claim 21, wherein the whole blood is treated with an anticoagulant selected from the group consisting of: sodium citrate, heparin, ethylenediaminetetraacetic acid (EDTA) and sodium oxylate.
24. The method of Claim 21, wherein the whole blood is fractionated by centrifuging the whole blood.
25. The method of Claim 21, wherein the purified red blood cells are lysed mechanically.
26. The method of Claim 21, wherein the whole blood is bovine whole blood.
27. A method of forming a lysate of purified red blood cells for use in a hemoglobin blood substitute, comprising the steps;
a) separating defibrinated whole bovine blood by centrifugation, whereby a red blood cell fraction and a liquid fraction are formed;
b) diafiltering the red blood cell fraction to thereby form purified red blood cells;
and c) mechanically lysing the purified red blood cells, thereby forming the lysate of purified red blood cells.
CA2476177A 2002-02-28 2002-02-28 Purification of red blood cells by separation and diafiltration Expired - Fee Related CA2476177C (en)

Applications Claiming Priority (1)

Application Number Priority Date Filing Date Title
PCT/US2002/006799 WO2003074077A1 (en) 2002-02-28 2002-02-28 Purification of red blood cells by separation and diafiltration

Publications (2)

Publication Number Publication Date
CA2476177A1 true CA2476177A1 (en) 2003-09-12
CA2476177C CA2476177C (en) 2011-06-14

Family

ID=27787374

Family Applications (1)

Application Number Title Priority Date Filing Date
CA2476177A Expired - Fee Related CA2476177C (en) 2002-02-28 2002-02-28 Purification of red blood cells by separation and diafiltration

Country Status (6)

Country Link
EP (1) EP1478388A1 (en)
JP (1) JP2006500317A (en)
CN (1) CN100348267C (en)
AU (1) AU2002245600B2 (en)
CA (1) CA2476177C (en)
WO (1) WO2003074077A1 (en)

Families Citing this family (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
PE20150961A1 (en) * 2009-05-14 2015-07-04 Biotechnology Inst I Mas D Sl EXTRACTION DEVICE FOR THE EXECUTION OF A METHOD OF PREPARATION OF AT LEAST ONE COMPOUND FROM BLOOD
US9550016B2 (en) * 2014-01-20 2017-01-24 Halcyon Biomedical, Incorporated Passive separation of whole blood
WO2015130778A1 (en) * 2014-02-25 2015-09-03 Biomet Biologics, Llc Cell filter separation system
GB201421013D0 (en) * 2014-11-26 2015-01-07 Turzi Antoine New standardizations & medical devices for the preparation of platelet rich plasma (PRP) or bone marrow centrate (BMC)

Family Cites Families (9)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US5084558A (en) 1987-10-13 1992-01-28 Biopure Corporation Extra pure semi-synthetic blood substitute
CA1312009C (en) 1986-11-10 1992-12-29 Carl W. Rausch Extra pure semi-synthetic blood substitute
US5854209A (en) 1995-03-23 1998-12-29 Biopure Corporation Method for oxygenating tissue having reduced red blood cell flow
EP0804244A1 (en) * 1993-03-26 1997-11-05 Biorelease Technologies, Inc. Compositions including heme-containing proteins and methods relating thereto
US5895810A (en) 1995-03-23 1999-04-20 Biopure Corporation Stable polymerized hemoglobin and use thereof
US5691452A (en) 1995-03-23 1997-11-25 Biopure Corporation Method for preserving a hemoglobin blood substitute
AU705225B2 (en) * 1995-03-23 1999-05-20 Biopure Corporation Stable polymerized hemoglobin blood substitute
US5691453A (en) 1995-06-07 1997-11-25 Biopure Corporation Separation of polymerized hemoglobin from unpolymerized hemoglobin on hydroxyapatite using HPLC
US6518010B2 (en) * 2001-02-28 2003-02-11 Biopure Corporation Use of defibrinated blood for manufacture of a hemoglobin-based oxygen carrier

Also Published As

Publication number Publication date
EP1478388A1 (en) 2004-11-24
CN100348267C (en) 2007-11-14
CA2476177C (en) 2011-06-14
WO2003074077A1 (en) 2003-09-12
CN1622823A (en) 2005-06-01
JP2006500317A (en) 2006-01-05
AU2002245600B2 (en) 2006-12-21
AU2002245600A1 (en) 2003-09-16

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