EP0379534A1 - Verfahren zur reinigung von vernetztem hämoglobin - Google Patents
Verfahren zur reinigung von vernetztem hämoglobinInfo
- Publication number
- EP0379534A1 EP0379534A1 EP89906273A EP89906273A EP0379534A1 EP 0379534 A1 EP0379534 A1 EP 0379534A1 EP 89906273 A EP89906273 A EP 89906273A EP 89906273 A EP89906273 A EP 89906273A EP 0379534 A1 EP0379534 A1 EP 0379534A1
- Authority
- EP
- European Patent Office
- Prior art keywords
- hemoglobin
- cross
- linked
- mixture
- heating
- 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.)
- Withdrawn
Links
- 108010054147 Hemoglobins Proteins 0.000 title claims abstract description 102
- 102000001554 Hemoglobins Human genes 0.000 title claims abstract description 102
- 238000000034 method Methods 0.000 title claims abstract description 28
- 239000000203 mixture Substances 0.000 claims abstract description 18
- 239000002244 precipitate Substances 0.000 claims abstract description 11
- 238000001556 precipitation Methods 0.000 claims abstract description 7
- 238000010438 heat treatment Methods 0.000 claims description 22
- 239000000243 solution Substances 0.000 claims description 19
- 238000006243 chemical reaction Methods 0.000 claims description 10
- 239000003638 chemical reducing agent Substances 0.000 claims description 10
- 108010061951 Methemoglobin Proteins 0.000 claims description 8
- QVGXLLKOCUKJST-UHFFFAOYSA-N atomic oxygen Chemical group [O] QVGXLLKOCUKJST-UHFFFAOYSA-N 0.000 claims description 6
- 239000001301 oxygen Substances 0.000 claims description 6
- 229910052760 oxygen Inorganic materials 0.000 claims description 6
- 239000007864 aqueous solution Substances 0.000 claims description 3
- 239000007789 gas Substances 0.000 claims description 3
- IJGRMHOSHXDMSA-UHFFFAOYSA-N Atomic nitrogen Chemical compound N#N IJGRMHOSHXDMSA-UHFFFAOYSA-N 0.000 description 12
- 239000000047 product Substances 0.000 description 8
- 239000011541 reaction mixture Substances 0.000 description 7
- XKRFYHLGVUSROY-UHFFFAOYSA-N Argon Chemical compound [Ar] XKRFYHLGVUSROY-UHFFFAOYSA-N 0.000 description 6
- 238000005119 centrifugation Methods 0.000 description 6
- 229910052757 nitrogen Inorganic materials 0.000 description 6
- 238000004132 cross linking Methods 0.000 description 5
- DHMQDGOQFOQNFH-UHFFFAOYSA-N Glycine Chemical compound NCC(O)=O DHMQDGOQFOQNFH-UHFFFAOYSA-N 0.000 description 4
- IMQLKJBTEOYOSI-GPIVLXJGSA-N Inositol-hexakisphosphate Chemical compound OP(O)(=O)O[C@H]1[C@H](OP(O)(O)=O)[C@@H](OP(O)(O)=O)[C@H](OP(O)(O)=O)[C@H](OP(O)(O)=O)[C@@H]1OP(O)(O)=O IMQLKJBTEOYOSI-GPIVLXJGSA-N 0.000 description 4
- IMQLKJBTEOYOSI-UHFFFAOYSA-N Phytic acid Natural products OP(O)(=O)OC1C(OP(O)(O)=O)C(OP(O)(O)=O)C(OP(O)(O)=O)C(OP(O)(O)=O)C1OP(O)(O)=O IMQLKJBTEOYOSI-UHFFFAOYSA-N 0.000 description 4
- 239000003633 blood substitute Substances 0.000 description 4
- 238000004587 chromatography analysis Methods 0.000 description 4
- 238000002474 experimental method Methods 0.000 description 4
- 238000001914 filtration Methods 0.000 description 4
- 239000012528 membrane Substances 0.000 description 4
- 229940068041 phytic acid Drugs 0.000 description 4
- HEMHJVSKTPXQMS-UHFFFAOYSA-M Sodium hydroxide Chemical compound [OH-].[Na+] HEMHJVSKTPXQMS-UHFFFAOYSA-M 0.000 description 3
- 229910052786 argon Inorganic materials 0.000 description 3
- GRWZHXKQBITJKP-UHFFFAOYSA-L dithionite(2-) Chemical compound [O-]S(=O)S([O-])=O GRWZHXKQBITJKP-UHFFFAOYSA-L 0.000 description 3
- 210000003743 erythrocyte Anatomy 0.000 description 3
- RWSXRVCMGQZWBV-WDSKDSINSA-N glutathione Chemical compound OC(=O)[C@@H](N)CCC(=O)N[C@@H](CS)C(=O)NCC(O)=O RWSXRVCMGQZWBV-WDSKDSINSA-N 0.000 description 3
- 239000012535 impurity Substances 0.000 description 3
- 108010001708 stroma free hemoglobin Proteins 0.000 description 3
- INGWEZCOABYORO-UHFFFAOYSA-N 2-(furan-2-yl)-7-methyl-1h-1,8-naphthyridin-4-one Chemical compound N=1C2=NC(C)=CC=C2C(O)=CC=1C1=CC=CO1 INGWEZCOABYORO-UHFFFAOYSA-N 0.000 description 2
- PREOBXYMXLETCA-UHFFFAOYSA-N 2-[4-(2-carboxyphenoxy)-4-oxobutanoyl]oxybenzoic acid Chemical compound OC(=O)C1=CC=CC=C1OC(=O)CCC(=O)OC1=CC=CC=C1C(O)=O PREOBXYMXLETCA-UHFFFAOYSA-N 0.000 description 2
- CIWBSHSKHKDKBQ-JLAZNSOCSA-N Ascorbic acid Chemical compound OC[C@H](O)[C@H]1OC(=O)C(O)=C1O CIWBSHSKHKDKBQ-JLAZNSOCSA-N 0.000 description 2
- SXRSQZLOMIGNAQ-UHFFFAOYSA-N Glutaraldehyde Chemical compound O=CCCCC=O SXRSQZLOMIGNAQ-UHFFFAOYSA-N 0.000 description 2
- 108010024636 Glutathione Proteins 0.000 description 2
- 239000004471 Glycine Substances 0.000 description 2
- 229920005654 Sephadex Polymers 0.000 description 2
- 239000012507 Sephadex™ Substances 0.000 description 2
- 229910052783 alkali metal Inorganic materials 0.000 description 2
- 150000001340 alkali metals Chemical class 0.000 description 2
- 239000008280 blood Substances 0.000 description 2
- 239000003431 cross linking reagent Substances 0.000 description 2
- 108010036302 hemoglobin AS Proteins 0.000 description 2
- 239000011261 inert gas Substances 0.000 description 2
- 238000004255 ion exchange chromatography Methods 0.000 description 2
- 210000003734 kidney Anatomy 0.000 description 2
- 235000004252 protein component Nutrition 0.000 description 2
- 238000010926 purge Methods 0.000 description 2
- 238000011160 research Methods 0.000 description 2
- -1 sulfoxy compounds Chemical class 0.000 description 2
- 239000006228 supernatant Substances 0.000 description 2
- INZBQWPDNWVYFR-OWOJBTEDSA-N 3,5-dibromo-2-[(e)-4-(2,4-dibromo-6-carboxyphenoxy)-4-oxobut-2-enoyl]oxybenzoic acid Chemical compound OC(=O)C1=CC(Br)=CC(Br)=C1OC(=O)\C=C\C(=O)OC1=C(Br)C=C(Br)C=C1C(O)=O INZBQWPDNWVYFR-OWOJBTEDSA-N 0.000 description 1
- BFBZHSOXKROMBG-UHFFFAOYSA-N 3,5-dibromo-2-hydroxybenzoic acid Chemical compound OC(=O)C1=CC(Br)=CC(Br)=C1O BFBZHSOXKROMBG-UHFFFAOYSA-N 0.000 description 1
- 208000030507 AIDS Diseases 0.000 description 1
- 229920002307 Dextran Polymers 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
- 239000004201 L-cysteine Substances 0.000 description 1
- 241001465754 Metazoa Species 0.000 description 1
- 239000002202 Polyethylene glycol Substances 0.000 description 1
- ZLMJMSJWJFRBEC-UHFFFAOYSA-N Potassium Chemical compound [K] ZLMJMSJWJFRBEC-UHFFFAOYSA-N 0.000 description 1
- LSNNMFCWUKXFEE-UHFFFAOYSA-N Sulfurous acid Chemical class OS(O)=O LSNNMFCWUKXFEE-UHFFFAOYSA-N 0.000 description 1
- 108010058907 Tiopronin Proteins 0.000 description 1
- 150000001450 anions Chemical class 0.000 description 1
- 229940072107 ascorbate Drugs 0.000 description 1
- 235000010323 ascorbic acid Nutrition 0.000 description 1
- 239000011668 ascorbic acid Substances 0.000 description 1
- 230000005540 biological transmission Effects 0.000 description 1
- OWMVSZAMULFTJU-UHFFFAOYSA-N bis-tris Chemical compound OCCN(CCO)C(CO)(CO)CO OWMVSZAMULFTJU-UHFFFAOYSA-N 0.000 description 1
- 210000004369 blood Anatomy 0.000 description 1
- 210000000601 blood cell Anatomy 0.000 description 1
- 239000000872 buffer Substances 0.000 description 1
- 239000008366 buffered solution Substances 0.000 description 1
- 239000006227 byproduct Substances 0.000 description 1
- 230000015556 catabolic process Effects 0.000 description 1
- 239000003153 chemical reaction reagent Substances 0.000 description 1
- 239000003795 chemical substances by application Substances 0.000 description 1
- 230000009089 cytolysis Effects 0.000 description 1
- 238000007872 degassing Methods 0.000 description 1
- 238000006731 degradation reaction Methods 0.000 description 1
- 238000011161 development Methods 0.000 description 1
- 238000011026 diafiltration Methods 0.000 description 1
- 108010074807 diaspirin-cross-linked hemoglobin Proteins 0.000 description 1
- 239000012153 distilled water Substances 0.000 description 1
- VHJLVAABSRFDPM-QWWZWVQMSA-N dithiothreitol Chemical compound SC[C@@H](O)[C@H](O)CS VHJLVAABSRFDPM-QWWZWVQMSA-N 0.000 description 1
- 239000000975 dye Substances 0.000 description 1
- 230000029142 excretion Effects 0.000 description 1
- 239000012467 final product Substances 0.000 description 1
- 238000011010 flushing procedure Methods 0.000 description 1
- 238000005187 foaming Methods 0.000 description 1
- 238000002523 gelfiltration Methods 0.000 description 1
- 229960003180 glutathione Drugs 0.000 description 1
- 208000006454 hepatitis Diseases 0.000 description 1
- 231100000283 hepatitis Toxicity 0.000 description 1
- 238000004128 high performance liquid chromatography Methods 0.000 description 1
- 238000001802 infusion Methods 0.000 description 1
- 150000002500 ions Chemical class 0.000 description 1
- 230000007774 longterm Effects 0.000 description 1
- 238000004519 manufacturing process Methods 0.000 description 1
- 231100000252 nontoxic Toxicity 0.000 description 1
- 230000003000 nontoxic effect Effects 0.000 description 1
- 239000007800 oxidant agent Substances 0.000 description 1
- 229920001223 polyethylene glycol Polymers 0.000 description 1
- 229910052700 potassium Inorganic materials 0.000 description 1
- 239000011591 potassium Substances 0.000 description 1
- 230000001376 precipitating effect Effects 0.000 description 1
- 235000018102 proteins Nutrition 0.000 description 1
- 102000004169 proteins and genes Human genes 0.000 description 1
- 108090000623 proteins and genes Proteins 0.000 description 1
- 238000000746 purification Methods 0.000 description 1
- 150000003839 salts Chemical class 0.000 description 1
- 238000000926 separation method Methods 0.000 description 1
- 150000003384 small molecules Chemical class 0.000 description 1
- 229910052708 sodium Inorganic materials 0.000 description 1
- 239000011734 sodium Substances 0.000 description 1
- JVBXVOWTABLYPX-UHFFFAOYSA-L sodium dithionite Chemical compound [Na+].[Na+].[O-]S(=O)S([O-])=O JVBXVOWTABLYPX-UHFFFAOYSA-L 0.000 description 1
- 239000012064 sodium phosphate buffer Substances 0.000 description 1
- 238000003860 storage Methods 0.000 description 1
- LSNNMFCWUKXFEE-UHFFFAOYSA-L sulfite Chemical class [O-]S([O-])=O LSNNMFCWUKXFEE-UHFFFAOYSA-L 0.000 description 1
- 239000000725 suspension Substances 0.000 description 1
- 125000003396 thiol group Chemical group [H]S* 0.000 description 1
- 231100000419 toxicity Toxicity 0.000 description 1
- 230000001988 toxicity Effects 0.000 description 1
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Chemical compound O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 1
Classifications
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K14/00—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
- C07K14/795—Porphyrin- or corrin-ring-containing peptides
- C07K14/805—Haemoglobins; Myoglobins
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K38/00—Medicinal preparations containing peptides
Definitions
- red blood cells as an oxygen carrier in the circulatory system of a living patient. This research has been prompted by the fact that these substitutes offer several potential advantages over the use of whole blood or red blood cells. As one advantage, an artificial oxygen carrier would not require typing and cross-matching as is the case for red blood cells. As another advantage, an artificial oxygen carrier would be very likely free of the risk of AIDS or hepatitis transmission.
- a third advantage of red cell substitutes is the potential for long term storage. Early experiments in this field evaluated the use of stroma-free hemoglobin as a red cell substitute. When stroma-free hemoglobin is infused intravenously into animals, its serves briefly as an oxygen carrier in the circulatory system.
- a method for purifying cross-linked hemoglobin which is mixed with non-cross-linked hemoglobin.
- the cross-linked hemoglobin is heated at a temperature of about 60 degrees to 85 degrees C. for a time sufficient to cause the precipitation of a substantial amount of the non-cross-linked hemoglobin (and frequently other impurities as well) present, without precipitating major amounts of cross-linked hemoglobin.
- the precipitate formed from the cross-linked hemoglobin typically b.y centrifugation or filtration.
- the cross-linked hemoglobin is processed in accordance with this invention at a pH of 6.5 to 9, most preferably a pH of 7 to 8, and specifically a pH of about 7.5.
- the duration of the heating step is preferably substantially one to six hours.
- the hemoglobin may be maintained in the substantially deoxygenated state during heating. This can be accomplished by various solution degassing procedures. These include, but are not limited to, sparging with inert gases; circulation through membrane gas exchange devices; and exposing hemoglobin solutions to a vacuum. The suitability of such procedures will be limited by the extent that they promote degradation of hemoglobin, for example through foaming, acidi ⁇ fication, or the like. For example, one may pass hemoglobin through a membrane oxygenator, for example a model number 08-2A membrane oxygenator of Sci-Med Life Systems Inc. of Minneapolis, Minnesota, with the gas channel of the oxygenator being filled with flowing nitrogen or argon.
- a membrane oxygenator for example a model number 08-2A membrane oxygenator of Sci-Med Life Systems Inc. of Minneapolis, Minnesota, with the gas channel of the oxygenator being filled with flowing nitrogen or argon.
- hemoglobin may be deoxygenated, followed by heating in accordance with this invention in a sealed, oxygen- free container to prevent reoxygenation.
- hemoglobin solutions may be sparged with an oxygen-free inert gas such as nitrogen or argon making use, for example, of a well known bubble-type oxygenator.
- the hemoglobin may also be maintained in its deoxy form using an appropriate reducing agent.
- a reducing agent generally is a chemireductant which should be physiologically acceptable and will typically have a reducing potential greater than or more effective than ascorbate against hemoglobin.
- Reduced redox dyes and sulfhydryl or sulfoxy compounds include many acceptable reducing agents. Suitable reducing agents also may include alkali metal (e.g. sodium or potassium) dithionites, bisulfites, metabisulfites, or sulfites. Other soluble, non-toxic salts of such anions may be candidates for use as well. Additionally, reduced glutathione or dithiothreitol may be used as well.
- the quantity of reducing agent to be included in the hemoglobin composition may vary, depending upon the reducing strength of the agent, the quantity of hemoglobin present, the temperature and duration of heating exposure, the presence of oxidizing agents, and other factors as will be apparent to the skilled artisan. Accordingly, the optimal concentration will be determined by routine experiments, for example by following the changes in hemoglobin composition as determined by ion exchange high performance liquid chromatography during the heating process. Dithionite may typically be used in a concentration of about 10 to 100 M in hemoglobin solutions, preferably about from 20 to 40 mM (expressed in terms of mM per litre). Other preferred reducing agents which may be used include glutathione, N- ⁇ cetyl-L-cysteine , and N- 2-mercapto-propionyl glycine.
- the deoxygenated solution containing both cross-linked and non-cross-linked hemoglobin and its impurities at a concentration of 1-10 grams per deciliter and a solution pH of 7 to 8.
- the heating may be at about 65 or 70 to 80 degrees C. for about 1-6 hours under an inert atmosphere of typically nitrogen or argon.
- a precipitation of non-cross-linked hemoglobin will take place, along with other by ⁇ products of the reaction in which the cross-linked hemoglobin was formed.
- the resulting precipitate can be removed by centrifugation and/or filtration, while a large percentage of the cross-linked hemoglobin does not precipitate and remains in solution.
- the hemoglobin may be maintained in substantially the oxygenated state during heating.
- a buffered solution of hemoglobin at concentration and pH preferably in the ranges described above may be heated preferably from about 60 to 75 degrees C, typically at about 65 degrees C, for a period of time of about 1 to 6 hours, for example one and one-half hours.
- the non-cross-linked hemoglobin precipitates from solution while the cross-linked hemoglobin remains substantially in dissolved form. Then, filtration and/or centrifugation may take place to remove the precipitate, composed primarily of non-cross-linked hemoglobin and other protein impurities.
- reagents which may be used to crosslink non-cross-linked hemoglobin may be glutaraldehyde, dextran, polyethylene glycol, and the like, with specific processes for producing cross- linked hemoglobin being as specifically described in the patents cited above.
- the cross- linked hemoglobin may be prepared as described in Walder U.S. Patent No. 4,600,531.
- a crude reaction mixture of di ⁇ spirin cross- linked hemoglobin was prepared by adding 1.5 equivalents of dibromosalicyl-bis-fumarate (DBBF) to a deoxygenated solution containing 3 g/dL stroma-free hemoglobin, 10 mM sodium phosphate buffer, pH 7.0, and 10 equivalents of inositol hexaphosphate (IHP). The solution was stirred at 37 degrees C. for 2 hours. Several aliquots of this solution were removed, the pH adjusted to 7.4, and the aliquots deoxygenated by repeated, alternating exposure to vacuum and nitrogen by flushing and evacuating the hemoglobin in a small vessel six or seven times. The aliquots were then heated at 70 degrees C.
- DBBF dibromosalicyl-bis-fumarate
- a crude reaction mixture of diaspirin cross- linked hemoglobin was prepared as described in Example 1 and the solution rendered free of ions such as IHP, glycine, and 3,5-dibromosalicylate by diafiltration and chromatography on a Sephadex G-25 column.
- the hemoglobin-containing eluate was adjusted to a pH of 7.4, deoxygenated, and aliquots heated at 80 degrees C. for up to two hours.
- Samples were analyzed for hemoglobin content as described in Example 1.
- Table 2 demonstrate that the removal of one or more of the small molecules present in crude reaction mixtures results in enhanced thermal stability of hemoglobins in general, but that selective precipitation of the unmodified molecules is still possible by increasing the temperature to 80 degrees C.
- Hemoglobin was prepared from outdated blood by hypotonic lysis with distilled water. Stroma was removed by centrifugation of the suspension at 35000 x g for one hour.
- the cross-linking reaction was performed in bis-tris buffer, pH 7.2, under anaerobic conditions established by purging with nitrogen.
- the solution contained 1 mM hemoglobin and 5mM inositol hexaphosphate. After adding 1.5 equivalents of DBBF the reaction was allowed to proceed for 2 hours at 37 degrees C and then stopped by the addition of an equal volume of 2 M glycine adjusted to a pH of 8.0 with NaOH.
- the ratio of cross-linked product to unmodified hemoglobin as determined by analytical isoelectic focusing was 4:1.
- the sample was oxygenated with room air and then heated to 65 degrees C for 1.5 hours. This lead to precipitation of 32% of the total hemoglobin presen including all of the unmodified hemoglobin. The hemoglobin remaining in the supernatant contained 66% methemoglobin. The precipitated hemoglobin was removed by centrifugation and filtration through a sterile 0.22 micron pore-size membrane. The sample was then cooled to 4 degrees C, deoxygenated, and sodium dithionite added to a final concentration of 40 mM in order to reduce the methemoglobin back to the unoxidized form.
- the reaction was allowed to proceed for five minutes, and the excess dithionite was subsequently removed by gel filtration over a Sephadex G-25 column maintained under anaerobic conditions by purging the buffer with nitrogen.
- the final product contained 5% of the hemoglobin in the met form.
Landscapes
- Chemical & Material Sciences (AREA)
- Organic Chemistry (AREA)
- Health & Medical Sciences (AREA)
- General Health & Medical Sciences (AREA)
- Biochemistry (AREA)
- Biophysics (AREA)
- Life Sciences & Earth Sciences (AREA)
- Genetics & Genomics (AREA)
- Medicinal Chemistry (AREA)
- Molecular Biology (AREA)
- Proteomics, Peptides & Aminoacids (AREA)
- Gastroenterology & Hepatology (AREA)
- Medicines That Contain Protein Lipid Enzymes And Other Medicines (AREA)
- Peptides Or Proteins (AREA)
- Medicines Containing Material From Animals Or Micro-Organisms (AREA)
Applications Claiming Priority (2)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| US20734688A | 1988-06-15 | 1988-06-15 | |
| US207346 | 1988-06-15 |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| EP0379534A1 true EP0379534A1 (de) | 1990-08-01 |
| EP0379534A4 EP0379534A4 (en) | 1991-03-13 |
Family
ID=22770150
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| EP19890906273 Withdrawn EP0379534A4 (en) | 1988-06-15 | 1989-04-10 | Method of purifying cross-linked hemoglobin |
Country Status (4)
| Country | Link |
|---|---|
| EP (1) | EP0379534A4 (de) |
| JP (1) | JP2592973B2 (de) |
| IE (1) | IE891251L (de) |
| WO (1) | WO1989012456A1 (de) |
Families Citing this family (21)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US5955581A (en) * | 1986-11-10 | 1999-09-21 | Biopure Corporation | Method for producing a stable polymerized hemoglobin blood-substitute |
| US5753616A (en) * | 1986-11-10 | 1998-05-19 | Biopure Corporation | Method for producing a stable polymerized hemoglobin blood-substitute |
| US5840851A (en) * | 1993-07-23 | 1998-11-24 | Plomer; J. Jeffrey | Purification of hemoglobin |
| US6242417B1 (en) | 1994-03-08 | 2001-06-05 | Somatogen, Inc. | Stabilized compositions containing hemoglobin |
| US5631219A (en) * | 1994-03-08 | 1997-05-20 | Somatogen, Inc. | Method of stimulating hematopoiesis with hemoglobin |
| US6288027B1 (en) | 1995-03-23 | 2001-09-11 | Biopure Corporation | Preserving a hemoglobin blood substitute with a transparent overwrap |
| US5895810A (en) * | 1995-03-23 | 1999-04-20 | Biopure Corporation | Stable polymerized hemoglobin and use thereof |
| US6150507A (en) * | 1995-03-23 | 2000-11-21 | Biopure Corporation | Method for producing a purified hemoglobin product |
| US6271351B1 (en) | 1995-03-23 | 2001-08-07 | Biopure Corporation | Method for preserving a hemoglobin blood substitute |
| ES2179188T3 (es) * | 1995-03-23 | 2003-01-16 | Biopure Corp | Sustituto de la sangre a base de hemoglobina polimerizada estable. |
| US6610832B1 (en) | 1995-03-23 | 2003-08-26 | Biopure Corporation | Preserving a hemoglobin blood substitute with a transparent overwrap |
| US5691452A (en) * | 1995-03-23 | 1997-11-25 | Biopure Corporation | Method for preserving a hemoglobin blood substitute |
| US5741894A (en) * | 1995-09-22 | 1998-04-21 | Baxter International, Inc. | Preparation of pharmaceutical grade hemoglobins by heat treatment in partially oxygenated form |
| EP0863918A1 (de) * | 1995-11-30 | 1998-09-16 | Somatogen Inc. | Verfahren zur kontrollierung der funktionalität während der vernetzung von hämoglobine |
| US6518010B2 (en) | 2001-02-28 | 2003-02-11 | Biopure Corporation | Use of defibrinated blood for manufacture of a hemoglobin-based oxygen carrier |
| CA2444590C (en) | 2001-04-18 | 2009-12-01 | Robert L. Mcginnis | Stabilized hemoglobin solutions |
| US7001715B2 (en) | 2002-02-28 | 2006-02-21 | Biopure Corporation | Purification of red blood cells by separation and diafiltration |
| JP2009524436A (ja) | 2006-01-24 | 2009-07-02 | ノースフィールド ラボラトリーズ、インコーポレイテッド | 重合したヘモグロビンの培地、並びに島細胞の単離及び移植におけるその使用 |
| US7932356B1 (en) * | 2010-06-23 | 2011-04-26 | Bing Lou Wong | Method for the preparation of a heat stable oxygen carrier-containing pharmaceutical composition |
| US8084581B1 (en) | 2011-04-29 | 2011-12-27 | Bing Lou Wong | Method for removing unmodified hemoglobin from cross-linked hemoglobin solutions including polymeric hemoglobin with a high temperature short time heat treatment apparatus |
| US20130052232A1 (en) * | 2011-08-31 | 2013-02-28 | Bing Lou Wong | Method for the preparation of a heat stable oxygen carrier-containing composition facilating beta-beta cross-linking |
Family Cites Families (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JPS61501510A (ja) * | 1984-03-23 | 1986-07-24 | バクスタ−、トラベノ−ル、ラボラトリ−ズ、インコ−ポレイテッド | ビ−ルスリスクを減らしたヘモグロビンおよびその製造方法 |
-
1989
- 1989-04-10 JP JP1505829A patent/JP2592973B2/ja not_active Expired - Fee Related
- 1989-04-10 EP EP19890906273 patent/EP0379534A4/en not_active Withdrawn
- 1989-04-10 WO PCT/US1989/001489 patent/WO1989012456A1/en not_active Ceased
- 1989-04-18 IE IE125189A patent/IE891251L/xx unknown
Non-Patent Citations (2)
| Title |
|---|
| No further relevant documents have been disclosed. * |
| See also references of WO8912456A1 * |
Also Published As
| Publication number | Publication date |
|---|---|
| IE891251L (en) | 1989-12-15 |
| JPH03502580A (ja) | 1991-06-13 |
| JP2592973B2 (ja) | 1997-03-19 |
| EP0379534A4 (en) | 1991-03-13 |
| WO1989012456A1 (en) | 1989-12-28 |
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