US3395210A - Lyophilized diagnostic reagent for the determination of blood coagulation factors - Google Patents
Lyophilized diagnostic reagent for the determination of blood coagulation factors Download PDFInfo
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- US3395210A US3395210A US343210A US34321064A US3395210A US 3395210 A US3395210 A US 3395210A US 343210 A US343210 A US 343210A US 34321064 A US34321064 A US 34321064A US 3395210 A US3395210 A US 3395210A
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- factor
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- blood coagulation
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- 239000003153 chemical reaction reagent Substances 0.000 title description 17
- 102000015081 Blood Coagulation Factors Human genes 0.000 title description 7
- 108010039209 Blood Coagulation Factors Proteins 0.000 title description 7
- 239000003114 blood coagulation factor Substances 0.000 title description 7
- 229940019700 blood coagulation factors Drugs 0.000 title description 4
- 210000001772 blood platelet Anatomy 0.000 description 20
- 239000000203 mixture Substances 0.000 description 17
- 238000012360 testing method Methods 0.000 description 16
- 239000012190 activator Substances 0.000 description 13
- 108010080865 Factor XII Proteins 0.000 description 10
- 102000000429 Factor XII Human genes 0.000 description 10
- PGOHTUIFYSHAQG-LJSDBVFPSA-N (2S)-6-amino-2-[[(2S)-5-amino-2-[[(2S)-2-[[(2S)-2-[[(2S)-2-[[(2S)-4-amino-2-[[(2S)-2-[[(2S)-2-[[(2S)-2-[[(2S)-2-[[(2S)-5-amino-2-[[(2S)-5-amino-2-[[(2S)-2-[[(2S)-2-[[(2S)-2-[[(2S,3R)-2-[[(2S)-5-amino-2-[[(2S)-2-[[(2S)-2-[[(2S,3R)-2-[[(2S)-2-[[(2S)-2-[[(2S)-2-[[(2S)-2-[[(2S)-5-amino-2-[[(2S)-1-[(2S,3R)-2-[[(2S)-2-[[(2S)-2-[[(2R)-2-[[(2S)-2-[[(2S)-2-[[2-[[(2S)-2-[[(2S)-2-[[(2S)-2-[[(2S)-1-[(2S)-2-[[(2S)-2-[[(2S)-2-[[(2S)-2-amino-4-methylsulfanylbutanoyl]amino]-3-(1H-indol-3-yl)propanoyl]amino]-5-carbamimidamidopentanoyl]amino]propanoyl]pyrrolidine-2-carbonyl]amino]-3-methylbutanoyl]amino]-4-methylpentanoyl]amino]-4-methylpentanoyl]amino]acetyl]amino]-3-hydroxypropanoyl]amino]-4-methylpentanoyl]amino]-3-sulfanylpropanoyl]amino]-4-methylsulfanylbutanoyl]amino]-5-carbamimidamidopentanoyl]amino]-3-hydroxybutanoyl]pyrrolidine-2-carbonyl]amino]-5-oxopentanoyl]amino]-3-hydroxypropanoyl]amino]-3-hydroxypropanoyl]amino]-3-(1H-imidazol-5-yl)propanoyl]amino]-4-methylpentanoyl]amino]-3-hydroxybutanoyl]amino]-3-(1H-indol-3-yl)propanoyl]amino]-5-carbamimidamidopentanoyl]amino]-5-oxopentanoyl]amino]-3-hydroxybutanoyl]amino]-3-hydroxypropanoyl]amino]-3-carboxypropanoyl]amino]-3-hydroxypropanoyl]amino]-5-oxopentanoyl]amino]-5-oxopentanoyl]amino]-3-phenylpropanoyl]amino]-5-carbamimidamidopentanoyl]amino]-3-methylbutanoyl]amino]-4-methylpentanoyl]amino]-4-oxobutanoyl]amino]-5-carbamimidamidopentanoyl]amino]-3-(1H-indol-3-yl)propanoyl]amino]-4-carboxybutanoyl]amino]-5-oxopentanoyl]amino]hexanoic acid Chemical compound CSCC[C@H](N)C(=O)N[C@@H](Cc1c[nH]c2ccccc12)C(=O)N[C@@H](CCCNC(N)=N)C(=O)N[C@@H](C)C(=O)N1CCC[C@H]1C(=O)N[C@@H](C(C)C)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](CC(C)C)C(=O)NCC(=O)N[C@@H](CO)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](CS)C(=O)N[C@@H](CCSC)C(=O)N[C@@H](CCCNC(N)=N)C(=O)N[C@@H]([C@@H](C)O)C(=O)N1CCC[C@H]1C(=O)N[C@@H](CCC(N)=O)C(=O)N[C@@H](CO)C(=O)N[C@@H](CO)C(=O)N[C@@H](Cc1cnc[nH]1)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H]([C@@H](C)O)C(=O)N[C@@H](Cc1c[nH]c2ccccc12)C(=O)N[C@@H](CCCNC(N)=N)C(=O)N[C@@H](CCC(N)=O)C(=O)N[C@@H]([C@@H](C)O)C(=O)N[C@@H](CO)C(=O)N[C@@H](CC(O)=O)C(=O)N[C@@H](CO)C(=O)N[C@@H](CCC(N)=O)C(=O)N[C@@H](CCC(N)=O)C(=O)N[C@@H](Cc1ccccc1)C(=O)N[C@@H](CCCNC(N)=N)C(=O)N[C@@H](C(C)C)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](CC(N)=O)C(=O)N[C@@H](CCCNC(N)=N)C(=O)N[C@@H](Cc1c[nH]c2ccccc12)C(=O)N[C@@H](CCC(O)=O)C(=O)N[C@@H](CCC(N)=O)C(=O)N[C@@H](CCCCN)C(O)=O PGOHTUIFYSHAQG-LJSDBVFPSA-N 0.000 description 9
- 230000035602 clotting Effects 0.000 description 9
- 108010000499 Thromboplastin Proteins 0.000 description 8
- 102000002262 Thromboplastin Human genes 0.000 description 8
- 206010053567 Coagulopathies Diseases 0.000 description 7
- 238000000034 method Methods 0.000 description 6
- 239000002245 particle Substances 0.000 description 6
- 230000007246 mechanism Effects 0.000 description 5
- VYPSYNLAJGMNEJ-UHFFFAOYSA-N Silicium dioxide Chemical compound O=[Si]=O VYPSYNLAJGMNEJ-UHFFFAOYSA-N 0.000 description 4
- 210000004556 brain Anatomy 0.000 description 4
- 239000000284 extract Substances 0.000 description 4
- BHPQYMZQTOCNFJ-UHFFFAOYSA-N Calcium cation Chemical compound [Ca+2] BHPQYMZQTOCNFJ-UHFFFAOYSA-N 0.000 description 3
- 108010074864 Factor XI Proteins 0.000 description 3
- 102000009123 Fibrin Human genes 0.000 description 3
- 108010073385 Fibrin Proteins 0.000 description 3
- BWGVNKXGVNDBDI-UHFFFAOYSA-N Fibrin monomer Chemical compound CNC(=O)CNC(=O)CN BWGVNKXGVNDBDI-UHFFFAOYSA-N 0.000 description 3
- 241000283973 Oryctolagus cuniculus Species 0.000 description 3
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical group [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 description 3
- 230000005856 abnormality Effects 0.000 description 3
- 230000015572 biosynthetic process Effects 0.000 description 3
- 229910001424 calcium ion Inorganic materials 0.000 description 3
- 230000015271 coagulation Effects 0.000 description 3
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- 239000000375 suspending agent Substances 0.000 description 3
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- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Chemical compound O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 3
- 241000220479 Acacia Species 0.000 description 2
- 239000005995 Aluminium silicate Substances 0.000 description 2
- HEDRZPFGACZZDS-UHFFFAOYSA-N Chloroform Chemical compound ClC(Cl)Cl HEDRZPFGACZZDS-UHFFFAOYSA-N 0.000 description 2
- 102100022641 Coagulation factor IX Human genes 0.000 description 2
- 102100023804 Coagulation factor VII Human genes 0.000 description 2
- 102100026735 Coagulation factor VIII Human genes 0.000 description 2
- 102100030563 Coagulation factor XI Human genes 0.000 description 2
- 108010076282 Factor IX Proteins 0.000 description 2
- 108010023321 Factor VII Proteins 0.000 description 2
- 229920002907 Guar gum Polymers 0.000 description 2
- 229920000084 Gum arabic Polymers 0.000 description 2
- 101000911390 Homo sapiens Coagulation factor VIII Proteins 0.000 description 2
- VEXZGXHMUGYJMC-UHFFFAOYSA-N Hydrochloric acid Chemical compound Cl VEXZGXHMUGYJMC-UHFFFAOYSA-N 0.000 description 2
- 235000010643 Leucaena leucocephala Nutrition 0.000 description 2
- 108010094028 Prothrombin Proteins 0.000 description 2
- 102100027378 Prothrombin Human genes 0.000 description 2
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- 239000003708 ampul Substances 0.000 description 2
- 239000003125 aqueous solvent Substances 0.000 description 2
- 239000000440 bentonite Substances 0.000 description 2
- 229910000278 bentonite Inorganic materials 0.000 description 2
- SVPXDRXYRYOSEX-UHFFFAOYSA-N bentoquatam Chemical compound O.O=[Si]=O.O=[Al]O[Al]=O SVPXDRXYRYOSEX-UHFFFAOYSA-N 0.000 description 2
- 230000023555 blood coagulation Effects 0.000 description 2
- 239000002026 chloroform extract Substances 0.000 description 2
- 230000007812 deficiency Effects 0.000 description 2
- 230000002950 deficient Effects 0.000 description 2
- 238000003745 diagnosis Methods 0.000 description 2
- 230000000694 effects Effects 0.000 description 2
- 239000011521 glass Substances 0.000 description 2
- 239000000665 guar gum Substances 0.000 description 2
- 235000010417 guar gum Nutrition 0.000 description 2
- 229960002154 guar gum Drugs 0.000 description 2
- 230000003993 interaction Effects 0.000 description 2
- NLYAJNPCOHFWQQ-UHFFFAOYSA-N kaolin Chemical compound O.O.O=[Al]O[Si](=O)O[Si](=O)O[Al]=O NLYAJNPCOHFWQQ-UHFFFAOYSA-N 0.000 description 2
- 239000000463 material Substances 0.000 description 2
- 229940039716 prothrombin Drugs 0.000 description 2
- CFWRDBDJAOHXSH-SECBINFHSA-N 2-azaniumylethyl [(2r)-2,3-diacetyloxypropyl] phosphate Chemical compound CC(=O)OC[C@@H](OC(C)=O)COP(O)(=O)OCCN CFWRDBDJAOHXSH-SECBINFHSA-N 0.000 description 1
- 208000002004 Afibrinogenemia Diseases 0.000 description 1
- 241000416162 Astragalus gummifer Species 0.000 description 1
- UXVMQQNJUSDDNG-UHFFFAOYSA-L Calcium chloride Chemical compound [Cl-].[Cl-].[Ca+2] UXVMQQNJUSDDNG-UHFFFAOYSA-L 0.000 description 1
- 206010016075 Factor I deficiency Diseases 0.000 description 1
- 108010014172 Factor V Proteins 0.000 description 1
- 108010014173 Factor X Proteins 0.000 description 1
- 108010049003 Fibrinogen Proteins 0.000 description 1
- 102000008946 Fibrinogen Human genes 0.000 description 1
- 208000031220 Hemophilia Diseases 0.000 description 1
- 208000009292 Hemophilia A Diseases 0.000 description 1
- 208000032843 Hemorrhage Diseases 0.000 description 1
- 239000005909 Kieselgur Substances 0.000 description 1
- 241001465754 Metazoa Species 0.000 description 1
- 206010037549 Purpura Diseases 0.000 description 1
- 241001672981 Purpura Species 0.000 description 1
- 108090000190 Thrombin Proteins 0.000 description 1
- 229920001615 Tragacanth Polymers 0.000 description 1
- 230000002159 abnormal effect Effects 0.000 description 1
- 235000010489 acacia gum Nutrition 0.000 description 1
- 239000002253 acid Substances 0.000 description 1
- 230000004913 activation Effects 0.000 description 1
- 239000004480 active ingredient Substances 0.000 description 1
- 238000003556 assay Methods 0.000 description 1
- 239000000305 astragalus gummifer gum Substances 0.000 description 1
- 230000000740 bleeding effect Effects 0.000 description 1
- 210000005013 brain tissue Anatomy 0.000 description 1
- 239000007853 buffer solution Substances 0.000 description 1
- 238000007820 coagulation assay Methods 0.000 description 1
- 239000008367 deionised water Substances 0.000 description 1
- 229910021641 deionized water Inorganic materials 0.000 description 1
- 201000010099 disease Diseases 0.000 description 1
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 description 1
- 229940079593 drug Drugs 0.000 description 1
- 239000003814 drug Substances 0.000 description 1
- 239000000428 dust Substances 0.000 description 1
- 238000002474 experimental method Methods 0.000 description 1
- 229940012952 fibrinogen Drugs 0.000 description 1
- 238000004108 freeze drying Methods 0.000 description 1
- 208000031169 hemorrhagic disease Diseases 0.000 description 1
- 238000011835 investigation Methods 0.000 description 1
- 238000009533 lab test Methods 0.000 description 1
- 210000005171 mammalian brain Anatomy 0.000 description 1
- 238000004519 manufacturing process Methods 0.000 description 1
- 230000007935 neutral effect Effects 0.000 description 1
- 230000003449 preventive effect Effects 0.000 description 1
- 230000008569 process Effects 0.000 description 1
- 239000004576 sand Substances 0.000 description 1
- 239000011780 sodium chloride Substances 0.000 description 1
- RGHFKWPGWBFQLN-UHFFFAOYSA-M sodium;5,5-diethylpyrimidin-3-ide-2,4,6-trione Chemical compound [Na+].CCC1(CC)C([O-])=NC(=O)NC1=O RGHFKWPGWBFQLN-UHFFFAOYSA-M 0.000 description 1
- 239000000126 substance Substances 0.000 description 1
- 238000001356 surgical procedure Methods 0.000 description 1
- 208000011580 syndromic disease Diseases 0.000 description 1
- 238000010998 test method Methods 0.000 description 1
- 229960004072 thrombin Drugs 0.000 description 1
- 235000010487 tragacanth Nutrition 0.000 description 1
- 229940116362 tragacanth Drugs 0.000 description 1
Classifications
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12Q—MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
- C12Q1/00—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions
- C12Q1/56—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions involving blood clotting factors, e.g. involving thrombin, thromboplastin, fibrinogen
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
- G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
- G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
- G01N33/86—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving blood coagulating time or factors, or their receptors
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
- G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
- G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
- G01N33/92—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving lipids, e.g. cholesterol, lipoproteins, or their receptors
-
- Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
- Y10—TECHNICAL SUBJECTS COVERED BY FORMER USPC
- Y10T—TECHNICAL SUBJECTS COVERED BY FORMER US CLASSIFICATION
- Y10T436/00—Chemistry: analytical and immunological testing
- Y10T436/10—Composition for standardization, calibration, simulation, stabilization, preparation or preservation; processes of use in preparation for chemical testing
- Y10T436/101666—Particle count or volume standard or control [e.g., platelet count standards, etc.]
-
- Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
- Y10—TECHNICAL SUBJECTS COVERED BY FORMER USPC
- Y10T—TECHNICAL SUBJECTS COVERED BY FORMER US CLASSIFICATION
- Y10T436/00—Chemistry: analytical and immunological testing
- Y10T436/10—Composition for standardization, calibration, simulation, stabilization, preparation or preservation; processes of use in preparation for chemical testing
- Y10T436/109163—Inorganic standards or controls
Definitions
- a further object of this invention is to provide a platelet factor reagent containing an activator and which will be useful as a reagent for the accurate determination of all plasma coagulation factors except Factor VII.
- the platelet factor reagent may be obtained, for example, by extracting warm-blooded mammal brain tissue such as human or rabbit brain in accordance with the procedure described by Belland Alton in Nature 174:880- 881 (1954). To the brain extract thus obtained is then added an activator in particulate form suspended in a buffer system having a pH of about 7.5 and the resulting mixture is then lyophilized.
- the particulate substance employed as the activator should be inert, resuspend very rapidly and have a uniform particle size and shouldnot have too alkaline or too acid a reaction to avoid any interference with the assay.
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- Health & Medical Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Engineering & Computer Science (AREA)
- Chemical & Material Sciences (AREA)
- Molecular Biology (AREA)
- Hematology (AREA)
- Immunology (AREA)
- Biomedical Technology (AREA)
- Urology & Nephrology (AREA)
- Biochemistry (AREA)
- Biotechnology (AREA)
- Organic Chemistry (AREA)
- Microbiology (AREA)
- General Health & Medical Sciences (AREA)
- Physics & Mathematics (AREA)
- Analytical Chemistry (AREA)
- Biophysics (AREA)
- Wood Science & Technology (AREA)
- General Physics & Mathematics (AREA)
- Pathology (AREA)
- Food Science & Technology (AREA)
- Medicinal Chemistry (AREA)
- Zoology (AREA)
- Proteomics, Peptides & Aminoacids (AREA)
- Cell Biology (AREA)
- Endocrinology (AREA)
- Neurosurgery (AREA)
- Bioinformatics & Cheminformatics (AREA)
- General Engineering & Computer Science (AREA)
- Genetics & Genomics (AREA)
- Investigating Or Analysing Biological Materials (AREA)
- Measuring Or Testing Involving Enzymes Or Micro-Organisms (AREA)
- Medicines Containing Material From Animals Or Micro-Organisms (AREA)
Description
LYOPHILIZED DIAGNOSTIC REAGENT FOR THE DETERMINATION OF BLOOD COAGULATION FACTORS Jane G. Lenahan, Florham Park, and George E. Phillips,
Morristown, N.J., assignors to Warner-Lambert Pharmaceutical Company, Morris Plains, NJ a corporation of Delaware No Drawing. Filed Feb. 7, 1964, Ser. No. 343,210
2 Claims. (Cl. 424-2) ABSTRACT OF THE DISCLOSURE This invention discloses a lyophilized diagnostic reagent for' the determination of blood coagulation factors. Broadly, this composition comprises a blood platelet factor substitute lyophilized together with finely-divided, discrete, inert particles.
This invention relates to novel compositions of matter. More particularly, this invention relates to compositions comprising a blood platelet factor reagent which contains a Hageman Factor activator and which is valuable for use as a diagnostic aid for the determination of blood coagulation factors. This invention also includes within its scope a novel process for the production of these compositions as well as methods for their use.
The mechanism of blood coagulation is complex but according to Miale, Laboratory Medicine & Hematology (1962) published by the C. V. Mosby Company, this mechanism may be considered as phenomenon resulting from the interaction of four plasma components:
( 1) prothrombin (2) thromboplastin (3) thrombin, and (4) fibrinogen in the presence of calcium ion to form a clot or fibrin. If there are any deficiencies in these components or any defect in their interaction, such abnormalities will be reflected in an altered blood coagulation picture which may be manifested by a failure to clot, under prolongation of clotting time, poor clot retraction and so on. Clinically, these abnormalities are manifested by syndromes such as hemophilia, purpura, afibrinogenemia and the like. In order to facilitate the diagnosis of these diseases, many laboratory tests have been devised to detect and diagnose the defective factor or factors so that proper preventive measures may be initiated or prior warning obtained when surgery is contemplated. These tests include, for example, the Duke method to determine bleeding time, the Lee-White method for determining clotting or coagulation time, and the Quick method to determine prothrombin. Recently, the partial thromboplastin time test, described by Iangdell et al., J. Lab. & Clin. Med. 41:637, 1953, has been widely recognized as a test procedure for testing the coagulation mechanism as a whole. This test determines the defects which may exist in all plasma clotting factors except platelet factor and Factor VII. This test is based on the theory that plasma thromboplastin is formed in the presence of platelet factor and in the presence of the following plasma factors viz: antihemophilic factor (AHF), Plasma Thromboplastin Component (PTC), Factor X, Factor XI or Plasma Thromboplastin Antecedent (PTA), Hageman Factor or Factor XII, calcium ion and Factor V.
Deficiencies or abnormalities in any one of the above factors leads to abnormal thromboplastin formation and this is reflected in the overall clinical coagulation picture.
In carrying out the test of partial thromboplastin time, the patients plasma is mixed with a standardized reagent nited States Patent 3,395,210 Patented July 30, 1968 ice containing a component which functions in the presence of optimum calcium ion concentration as platelet factor and thereby acts as a substitute, and the clotting time observed is noted. The platelet factor substitute is generally obtained as described below from warm-blooded animal brain tissue and is essentially cephalin having platelet factor-like activity. While this test is a valuable tool for the study of the generation of plasma thromboplastin and the diagnosis of various hemorrhagic disorders due to defects at this level, it has been observed that this test is very sensitive to certain environmental conditions. Thus, for example, if one carries out the test in glassware which has uneven surfaces this tends to accelerate the clotting time whereas the use of glassware having smooth surfaces, on the other hand, tends to give a rather extended end point. Obviously, for the test to be meaningful, the end-point should be reproducible, sharp and not so readily influenced by such environmental factors.
Investigations into these phenomena and experiments with normal plasma having pre-determined clotting time have ascertained that the presence of dust particles or of scratched glassware surface is responsible for the activation of the Hageman factor in the plasma whereas in the absence of such foci the action of the Hageman factor is slowed and the clotting mechanism therefore takes longer to act. Thus, even though none of the plasma factors are deficient, the results observed are variable and it appears that Hageman factor still needs to be activated to insure proper clotting activity. Clearly, then, while a platelet factor reagent is required in the clotting mechanism, the composition should also contain an activator which while insensitive to environmental conditions will activate the Hageman factor present and thereby insure uniform clotting action.
Accordingly, a primary object of this invention is to provide a modified platelet factor reagent containing an added Hageman factor activator of predictable action.
A further object of this invention is to provide a platelet factor reagent containing an activator and which will be useful as a reagent for the accurate determination of all plasma coagulation factors except Factor VII.
Another object of this invention is to provide a stable platelet factor reagent system containing an activator which will activate Hageman factor and which when used in coagulation assays gives a high order of reproducibility.
Other objects and advantages of this invention will become more apparent from the following detailed description.
Broadly speaking, the novel composition of this invention is a lyophilized composition which contains as the active ingredients a platelet factor reagent in combination with a Hageman factor activator of precise and controlled activity.
The platelet factor reagent may be obtained, for example, by extracting warm-blooded mammal brain tissue such as human or rabbit brain in accordance with the procedure described by Belland Alton in Nature 174:880- 881 (1954). To the brain extract thus obtained is then added an activator in particulate form suspended in a buffer system having a pH of about 7.5 and the resulting mixture is then lyophilized. The particulate substance employed as the activator should be inert, resuspend very rapidly and have a uniform particle size and shouldnot have too alkaline or too acid a reaction to avoid any interference with the assay.
As examples of suitable activators which may be employed, there may be mentioned, for example, finelydivided diatomaceous earth, bentonite, kaolin, sand and powdered glass, and having a particle size of about from 2 to 20 microns.
keted commercially by Johns-Manville Co. as Celite 505 is preferred for use as the activator because it is inert, uniform in size, resuspends readily and is essentially neutral in reaction.
For optimum results, We mix about 0.5 to 3% by Weight of activator with each 100 ml. of the platelet factor extract. In order to provide a uniform suspension when the lyophilized material is reconstituted in'an aqueous solvent, it has been found to be advantageous to include about 0.5 to 2% by weight of a suspending agent in the mixture undergoing lyophilization. Suspending agents such as calcium-free acacia, calcium-free tragacanthgcalciumf free guar gum and the like are particularly advantageous.
In order to further illustrate this invention,the following examples are given:
Example 1 tissues are then extracted with approximately 50 to 100 cc. of chloroform. The chloroform extracts are carefully evaporated to give a residue of from about 300 to-400 mg. This residue when homogenized in about 125 ml. of aqueous isotonic sodium chloride solution forms the desired extract.
The aqueous rabbit brain extract thus formed is blended with 32.5 grams of Celite 505 in about 3.1 liters of an aqueous solvent medium comprising grams of calcium-free acacia gum, 21 grams sodium barbital, and 15 grams of sodium chloride to obtain a uniform suspension. The pH of the suspension is then adjusted to 7.5 with 10% aqueous hydrochloric acid. About 2.5 ml. of the resulting mixture is filled into individual vials or ampules and the material lyophilized, the lyophilized product constituting the desired blood platelet factor reagent.
Example 2 To employ the lyophilized blood platelet factor reagent composition obtained in accordance with Example 1, the lyophilized unit in an individual ampule is first reconstituted in about 2.5 ml.' of deionized wtter. An aliquot portion of the aqueous composition is mixed with an equal volume of 0.025 molar calcium chloride solution. A standard human plasma, such as Diagnostic Plasma marketed by Warner-Chilcott which is essentially a pooled lyophilized normal human plasma standardized to contain optimal concentrations of all clotting factors is reconstituted in deionized water. 0.1 ml. of this reconstituted aqueous plasma is blown into a test tube containing 0.2
ml. of the reconstituted platelet factor reagent containing (1) 36.5 seconds (2) 36.0 seconds 3 36.7 seconds When the platelet factor reagent bufwithout the activator, elapsed time for the first fibrin strand formation in the respective tubes is:
(l),83.5 seconds I H 1 6-8 seconds Z; (3)'72.1 seconds i (1) 71.6 seconds (2) 69.9 seconds 73.1 seconds Thesame test is repeated employing the same platelet factor reagent but without the activator. The'elapsed time for the first fibrin strand formation in the respective tubes is: I
(1) 180.2 seconds" (2) 200.8 seconds (3) 168.5 seconds The above results overwhelmingly indicate 'the :reproducibility, sharpness and accuracy of the test as well as the highly advantageous rapidity of the test when employing the novel compositions of this invention in the partial thromboplastin time test.
It is to be understood that the foregoing detailed'description is given merely by way of illustration and that many variations may be made therein without departing from the spirit of our invention. 7
Having described our invention, what we desire tosecure by Letters Patent is:
1. A lyophilized composition of matter suitable for reconstitution by addition of Water comprising blood platelet substitute selected from the group consisting of human and rabbit brain chloroform extracts in combination with 0.5 to 3 parts by weight of finely-divided disjcrete inert particles selected from the group consisting of kaolin, powdered glass, bentonite, and dia'tomaceous earth and a suspending agent selected from the group consisting of calcium-free acacia, tragacanth, and guar gum, said composition being buffered to a pH of about 7.5..
2. A composition as defined in claim 1, wherein said particles arefrom about 2 to 20 microns in size.
References Cited 'UNITED sTA Es PATENTS 3,179,567 .4/1965 Owren 16784.'5
OTHER REFERENCES Proctor: Am. J. Clin. Path, 36:3, September 1961, pp. 212-219.
p Struver: Am. J. Clin. Path, 38:5, November 1962, pp.
Hyland: Partial Thromboplastin Time (PTT) Test, folder, 2 pp. Hyland Labs, Los Angeles, Calif, January 1964. ALBERT T. MEYERS, Primar yEicamin er.
FAGLESQN, Assistant Exairiiner.
Priority Applications (6)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| US343210A US3395210A (en) | 1964-02-07 | 1964-02-07 | Lyophilized diagnostic reagent for the determination of blood coagulation factors |
| GB51454/64A GB1029329A (en) | 1964-02-07 | 1964-12-17 | Lyophilized reagent composition |
| DEW38292A DE1266998B (en) | 1964-02-07 | 1965-01-05 | Blood platelet factor reagent designed for use in blood clotting tests |
| DK22165AA DK111541B (en) | 1964-02-07 | 1965-01-15 | Diagnostic agent for determining blood coagulation factors. |
| FR2191A FR1441469A (en) | 1964-02-07 | 1965-01-15 | Lyophilized composition for the determination of blood coagulation factors |
| JP40002412A JPS5010925B1 (en) | 1964-02-07 | 1965-01-19 |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| US343210A US3395210A (en) | 1964-02-07 | 1964-02-07 | Lyophilized diagnostic reagent for the determination of blood coagulation factors |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| US3395210A true US3395210A (en) | 1968-07-30 |
Family
ID=23345144
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| US343210A Expired - Lifetime US3395210A (en) | 1964-02-07 | 1964-02-07 | Lyophilized diagnostic reagent for the determination of blood coagulation factors |
Country Status (6)
| Country | Link |
|---|---|
| US (1) | US3395210A (en) |
| JP (1) | JPS5010925B1 (en) |
| DE (1) | DE1266998B (en) |
| DK (1) | DK111541B (en) |
| FR (1) | FR1441469A (en) |
| GB (1) | GB1029329A (en) |
Cited By (19)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US3650698A (en) * | 1969-12-04 | 1972-03-21 | Technicon Corp | Apparatus for the automatic determination of the coagulation, aggregation and or flocculation, or the like, rates of fluids, and novel reaction intensifying agent for use therewith |
| US3839314A (en) * | 1971-06-29 | 1974-10-01 | Baxter Laboratories Inc | Clarification of blood serum and plasma using block copolymers of ethylene oxide and polyoxypropylene |
| US3893991A (en) * | 1973-04-05 | 1975-07-08 | Baxter Laboratories Inc | Clarification of blood serum and plasma using block copolymers of ethylene oxide and polyoxypropylene |
| US3893990A (en) * | 1973-04-05 | 1975-07-08 | Baxter Laboratories Inc | Clarification of blood serum and plasma using a block copolymer of ethylene oxide and polyoxypropylene |
| US3947378A (en) * | 1974-12-23 | 1976-03-30 | Warner-Lambert Company | Adsorbed plasma |
| US4038147A (en) * | 1976-06-15 | 1977-07-26 | Becton, Dickinson And Company | Method for detecting endotoxins |
| US4197088A (en) * | 1977-09-23 | 1980-04-08 | Akro-Medic Engineering, Inc. | Method for qualitative and quantitative determination of immunological reactions |
| US4210557A (en) * | 1978-12-15 | 1980-07-01 | Beckman Instruments, Inc. | Non-high density lipoprotein precipitant |
| US4360358A (en) * | 1980-03-07 | 1982-11-23 | Yash Sharma | Immunoassay with solid phase having coating containing blood platelet substitute |
| US4411518A (en) * | 1977-09-23 | 1983-10-25 | Gamma Biologicals, Inc. | Apparatus for use in qualitative determination of immunological reactions |
| WO1983004105A1 (en) * | 1982-05-07 | 1983-11-24 | Cooper Laboratories, Inc. | Clotting assay and reagent therefor |
| WO1985004426A1 (en) * | 1984-03-26 | 1985-10-10 | International Health Services | A method of determining the clotting time of blood and particulate reagents therefor |
| FR2583881A1 (en) * | 1985-06-21 | 1986-12-26 | Girolami Antoine | Method and solution for determining the Howell times and performing heparin tolerance tests |
| US4755461A (en) * | 1986-04-17 | 1988-07-05 | Bio/Data Corporation | Tableted blood plasma microconcentrated thromboplastin coagulation reagent |
| US4849340A (en) * | 1987-04-03 | 1989-07-18 | Cardiovascular Diagnostics, Inc. | Reaction system element and method for performing prothrombin time assay |
| US6632678B2 (en) | 2001-01-03 | 2003-10-14 | Sienco, Inc. | Method for performing activated clotting time test with reduced sensitivity to the presence of aprotinin and for assessing aprotinin sensitivity |
| US20120231485A1 (en) * | 2011-03-08 | 2012-09-13 | Oenundarson Pall Torfi | Method for monitoring anticoagulant therapy |
| US11604026B2 (en) | 2019-03-14 | 2023-03-14 | Terumo Bct Biotechnologies, Llc | Lyophilization loading tray assembly and system |
| US11634257B2 (en) | 2017-10-09 | 2023-04-25 | Terumo Bct Biotechnologies, Llc | Lyophilization container and method of using same |
Families Citing this family (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| GB2132759B (en) * | 1979-09-11 | 1985-01-09 | Terumo Corp | Blood collecting tube |
| DE3407280A1 (en) * | 1984-02-28 | 1985-09-19 | Gödecke AG, 1000 Berlin | TEST SET FOR DETERMINING THE ACTIVATED PARTIAL THROMBOPLASTIN TIME (PTT) WITH INCREASED HEPARINE SENSITIVITY |
| CN116165039B (en) * | 2023-02-11 | 2023-09-22 | 保定天岳生物工程有限公司 | Preparation method of rabbit brain tissue replacing traditional rabbit brain powder |
Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US3179567A (en) * | 1958-05-16 | 1965-04-20 | Owren Paul Arnor | Reagent and method for determining the coagulability of blood |
-
1964
- 1964-02-07 US US343210A patent/US3395210A/en not_active Expired - Lifetime
- 1964-12-17 GB GB51454/64A patent/GB1029329A/en not_active Expired
-
1965
- 1965-01-05 DE DEW38292A patent/DE1266998B/en active Pending
- 1965-01-15 FR FR2191A patent/FR1441469A/en not_active Expired
- 1965-01-15 DK DK22165AA patent/DK111541B/en unknown
- 1965-01-19 JP JP40002412A patent/JPS5010925B1/ja active Pending
Patent Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US3179567A (en) * | 1958-05-16 | 1965-04-20 | Owren Paul Arnor | Reagent and method for determining the coagulability of blood |
Cited By (28)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US3650698A (en) * | 1969-12-04 | 1972-03-21 | Technicon Corp | Apparatus for the automatic determination of the coagulation, aggregation and or flocculation, or the like, rates of fluids, and novel reaction intensifying agent for use therewith |
| US3839314A (en) * | 1971-06-29 | 1974-10-01 | Baxter Laboratories Inc | Clarification of blood serum and plasma using block copolymers of ethylene oxide and polyoxypropylene |
| US3893991A (en) * | 1973-04-05 | 1975-07-08 | Baxter Laboratories Inc | Clarification of blood serum and plasma using block copolymers of ethylene oxide and polyoxypropylene |
| US3893990A (en) * | 1973-04-05 | 1975-07-08 | Baxter Laboratories Inc | Clarification of blood serum and plasma using a block copolymer of ethylene oxide and polyoxypropylene |
| US3947378A (en) * | 1974-12-23 | 1976-03-30 | Warner-Lambert Company | Adsorbed plasma |
| US4038147A (en) * | 1976-06-15 | 1977-07-26 | Becton, Dickinson And Company | Method for detecting endotoxins |
| US4197088A (en) * | 1977-09-23 | 1980-04-08 | Akro-Medic Engineering, Inc. | Method for qualitative and quantitative determination of immunological reactions |
| US4411518A (en) * | 1977-09-23 | 1983-10-25 | Gamma Biologicals, Inc. | Apparatus for use in qualitative determination of immunological reactions |
| US4210557A (en) * | 1978-12-15 | 1980-07-01 | Beckman Instruments, Inc. | Non-high density lipoprotein precipitant |
| US4360358A (en) * | 1980-03-07 | 1982-11-23 | Yash Sharma | Immunoassay with solid phase having coating containing blood platelet substitute |
| WO1983004105A1 (en) * | 1982-05-07 | 1983-11-24 | Cooper Laboratories, Inc. | Clotting assay and reagent therefor |
| US4455377A (en) * | 1982-05-07 | 1984-06-19 | Cooper Laboratories, Inc. | Clotting assay and reagent therefor |
| WO1985004426A1 (en) * | 1984-03-26 | 1985-10-10 | International Health Services | A method of determining the clotting time of blood and particulate reagents therefor |
| FR2583881A1 (en) * | 1985-06-21 | 1986-12-26 | Girolami Antoine | Method and solution for determining the Howell times and performing heparin tolerance tests |
| US4755461A (en) * | 1986-04-17 | 1988-07-05 | Bio/Data Corporation | Tableted blood plasma microconcentrated thromboplastin coagulation reagent |
| US4849340A (en) * | 1987-04-03 | 1989-07-18 | Cardiovascular Diagnostics, Inc. | Reaction system element and method for performing prothrombin time assay |
| US6197494B1 (en) * | 1987-04-03 | 2001-03-06 | Cardiovascular Diagnostics, Inc. | Apparatus for performing assays on liquid samples accurately, rapidly and simply |
| US6632678B2 (en) | 2001-01-03 | 2003-10-14 | Sienco, Inc. | Method for performing activated clotting time test with reduced sensitivity to the presence of aprotinin and for assessing aprotinin sensitivity |
| US20120231485A1 (en) * | 2011-03-08 | 2012-09-13 | Oenundarson Pall Torfi | Method for monitoring anticoagulant therapy |
| US9234902B2 (en) * | 2011-03-08 | 2016-01-12 | Pall Torfi Önundarson | Method for monitoring anticoagulant therapy |
| US11634257B2 (en) | 2017-10-09 | 2023-04-25 | Terumo Bct Biotechnologies, Llc | Lyophilization container and method of using same |
| US11609042B2 (en) | 2019-03-14 | 2023-03-21 | Terumo Bct Biotechnologies, Llc | Multi-part lyophilization container and method of use |
| US11609043B2 (en) | 2019-03-14 | 2023-03-21 | Terumo Bct Biotechnologies, Llc | Lyophilization container fill fixture, system and method of use |
| US11604026B2 (en) | 2019-03-14 | 2023-03-14 | Terumo Bct Biotechnologies, Llc | Lyophilization loading tray assembly and system |
| US11740019B2 (en) | 2019-03-14 | 2023-08-29 | Terumo Bct Biotechnologies, Llc | Lyophilization loading tray assembly and system |
| US11747082B2 (en) | 2019-03-14 | 2023-09-05 | Terumo Bct Biotechnologies, Llc | Multi-part lyophilization container and method of use |
| US11815311B2 (en) | 2019-03-14 | 2023-11-14 | Terumo Bct Biotechnologies, Llc | Lyophilization container fill fixture, system and method of use |
| US11994343B2 (en) | 2019-03-14 | 2024-05-28 | Terumo Bct Biotechnologies, Llc | Multi-part lyophilization container and method of use |
Also Published As
| Publication number | Publication date |
|---|---|
| DE1266998B (en) | 1968-04-25 |
| DK111541B (en) | 1968-09-09 |
| FR1441469A (en) | 1966-06-10 |
| JPS5010925B1 (en) | 1975-04-25 |
| GB1029329A (en) | 1966-05-11 |
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