CA1226795A - Stabilized coagulation control products - Google Patents
Stabilized coagulation control productsInfo
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- CA1226795A CA1226795A CA000455086A CA455086A CA1226795A CA 1226795 A CA1226795 A CA 1226795A CA 000455086 A CA000455086 A CA 000455086A CA 455086 A CA455086 A CA 455086A CA 1226795 A CA1226795 A CA 1226795A
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Abstract
STABILIZED COAGULATION CONTROL PRODUCT
Abstract Reagents and methods for stabilized coagulation control reagents and plasma resin control reagents. In the most preferred embodiment, the coagulation control reagents of the present invention comprise substantially defibrinated plasma to which has been added an anticoagulation means, and plexiform stabilizing means in an amount sufficient so that it is present in a final concentration of about 3% to 10%. Specific clotting Factors may also be removed in accordance with the type of control reagent desired. A
most preferred embodiment employs lactose as the plexiform stabilizing means.
Abstract Reagents and methods for stabilized coagulation control reagents and plasma resin control reagents. In the most preferred embodiment, the coagulation control reagents of the present invention comprise substantially defibrinated plasma to which has been added an anticoagulation means, and plexiform stabilizing means in an amount sufficient so that it is present in a final concentration of about 3% to 10%. Specific clotting Factors may also be removed in accordance with the type of control reagent desired. A
most preferred embodiment employs lactose as the plexiform stabilizing means.
Description
US
Field of the Invention This invention relates to stabilized control products useful in the clinical environment and in particular relates to stabilized coagulation control reagents and plasma-renin control reagents.
Background of the Invention This invention is related to a cop ending Canadian apply-cation Serial No. 455,085 entitled "A Stabilized Multi-parameter Control Product" (OARED) filed May 25, 1984 by the inventor hereof.
Hemostasis is a complicated life saving procedure involve in blood vessels plasma, platelets, vessel walls, neural and hum oral activity all for the purpose of stanching bleeding. Although there has been much research, especially within the last few decades, concerning the process of coagulation specifically and hemostasis in general, there is much that is not completely understood particularly with regard to the specific details concern-in the interaction of contributing mechanisms. For instance it is now known that there are no less than 35 compounds which take part in the formation of a firm blood clot. Ostensibly, blood coagulation is thus one of thy more complicated chemical processes occurring within the body In fact, there are so many separate components within the scheme of coagulation that the International Committee on nomenclature for Blood Coagulation formed by the International Hematology Congress has adopted a series of names which have now come into general acceptance. The most common of these names (with generally employed OARED
alternatives) and factor designations include the following:
Factor I Fibrinogen Factor II Prothrombin Factor III Thromboplastin Factor IV Calcium Factor V Proaccelerin, labile factor Factor VI (no longer used) Factor VII Serum prothrombin conversion accelerator (SPA), stable factor Factor VIII Antihemophilic factor (AH) Factor IX Christmas factor, plasma thromboplastin component (PTC) Factor X Stuart Factor, Stuart-Prower factor Factor XI Plasma thromboplastin antecedent (PTA) Factor XII Hegemony factor Factor XIII Fibrin stabilizing factor Profibrinolysin Plasminogen Fibrinolysin Plasm in Diagnostic determination of the presence and quantitation of various of the above factors is important as many clinically relevant diseases associated with abnormal clotting are associated therewith. For instance, the absence of Factor VIII is the most frequent and serious cause of genetically determined clotting defects such as hemophilia A which has been recognized for over 2,000 years. A deficiency in plasma concentration of Factor IX
is generally associated with the-genetically determined defect responsible for hemophilia B. hemorrhagic disease results from deficiencies in Factors X and XI whereas deficiencies in Factor XII induce longer clotting times I
but still permit eventual clotting. Diseases of the liver or Vitamin K deficiencies have also been associated with abnormal levels of clotting Factors II, VII, IX r and X as these are predominantly produced by the liver. Other causes of abnormal clotting may be related to reduced platelet counts, ~hrombocytopenia, often associated with abnormal levels of clotting Factors II, VII, IX, and X as these are predominantly produced by the liver. Other causes of abnormal clotting may be related to reduced platelet counts, thrombocytopenia, often associated with pernicious anemia, certain drug therapies, irradiation or increased peripheral destruction by antibodies.
The clinical value of testing for coagulation times is not limited to the detection of genetic or pathological disease states but is also useful in the regulation of anticoagulant therapy. Anticoagulants typically inhibit the coagulation mechanism such as by the heparin mediated inhibition of Factor X by Antithrombin III. Deficiencies in the clotting mechanisms due to congenital defects, pathological conditions, or anti coagulation therapy have been discussed generally in Chapter 7, "Coagulation and Hemostasis" by Robert D. Languidly, In Clinical Diagnosis, Davidson and Henry, 1979.
As commonly understood, coagulation may occur by two path-ways, the so called intrinsic pathway and extrinsic path-way. The former is generally triggered by the presence of a surface (thought to activate Factor XII) and, with the presence of phospholipids and calcium, through a number of steps eventually stimulates the formation of a stabilized fibrin clot. The partial thromboplastin time (PUT) test typically measures the intrinsic pathway wherein most congenital deficiencies occur. Consequently this type of test serves as an excellent presurgical coagulation screening test; however, since the reagent employed in OARED
us this test is typically a platelet substitute, the PUT test does not measure platelet activity.
The extrinsic pathway is generally triggered by injury to tissue and the resultant exudation of tissue thromboplas-tin acts on Factors VII and X which, following a series of steps, thereafter forms a fibrin clot. The extrinsic pathway is generally tested by the so called one-stage prothrombin time test to thereby detect most of the factors depressed by oral anticoagulant drugs for antic-agulant therapy control. The thrombin time test typically measures the quantity and reactivity of fibrinogen. This test is a rapid, semi quantitative test and is consequently the test of choice for intravascular clotting and fibrinolysis analysis. The above is presented in review in accompanying Table I as well as a booklet entitled "Concentric Concepts of Coagulation 1971, 1975" available from Ortho Diagnostics, Inc., Raritan, New Jersey.
OARED
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With such attention being placed on the determination of a particular patient's coagulating ability and the presence or absence of specific factors, it is axiomatic that adequate controls must be available in order to ensure the proper operation of manual and automated methods designed to determine these factor levels and coagulation in general. Heretofore, such controls as were available have been typically characterized by relatively short shelf fifes both in the liquid and dry state due largely, in part, to the unstable nature of the ingredients or by their inability to withstand drying procedures such as lyophilization.
It us an object of the present invention to provide plasma-renin and coagulation control reagents having the appropriate levels of particular factors present therein in a stabilized format and capable of being lyophilized.
It is another object of the present invention to provide methods whereby coagulation control and plasma-renin control reagents may be stabilized It is another related object of the present invention to provide reagents in a stabilized format useful in manual and automated renal hypertensive testing and coagulation testing.
Brief Summary of the Invention In accordance with the objects and principles of the present invention, coagulation control reagents and plasma resin control reagents are provided which are substantially stabilized by the addition of plexiform stabilizing means. In the case of coagulation control reagents, anti coagulation means is added to substantially defibrinated plasma, whereby clotting components are held OARED
intact and residual fibrin formation is inhibited.
Additionally, plexiform stabilizing means, selected from the group consisting of sucrose, reducing monosaccharide sugars and reducing disaccharide sugars, is added so that S it is present in the final product in he range of about 3%-10% volume percent The preferred plexiform stabilizing means is selected from the group consisting of maltose, minutely, syllabus, glucose and lactose with the latter most being the most preferred. Additionally, and in accordance with the typo of coagulation control reagent desired, a specific clotting factor or factors may be removed such as Factor VIII and/or Factor I.
The plasma resin control reagent of the present invention comprises selected plasma to which has been added anti coagulation means whereby clotting components are held substantially intact, and plexiform stabilizing means similar to that described for the coagulation control reagent. The anti coagulation means is ideally chosen from eye group consisting of citrate, oxalate, ETA (ethylene Damon tetraacetic acid), heparin, and any combination of the foregoing.
The most preferred embodiments will have substantially all water removed by processes well known such as by lyophilization~
In another embodiment the present invention provides a method for producing a stabilized coagulation control reagent comprising the steps of: (a) providing whole blood; (b) removing substantially all cells from said whole blood to obtain plasma; (c) adding anti coagulation means whereby clotting components are held intact and fibrin formation is inhibited; (d) adjusting specific clotting Factor or Factors in accordance with the type of coagulation control reagent desired, (e) adding plexiform stabilizing means -pa-selected from the group consisting of sucrose, reducing monosaccharide sugars and reducing disaccharide sugars so that said plexiform stabilizing means is present in the range of about 3% to 10% final volume percent, In still another embodiment the present invention provides a stabilized plasma resin control resent useful for Anglo-tension testing comprising: (a) plasma having a known resin level; (b) anti coagulation means whereby clotting components are held intact and fibrin formation is substantially inhibited; and (c) plexiform stabilizing means selected from the group consisting of reducing monosaccharide sugars and reducing disaccharide sugars whereby said plexiform stabilizing means is present in the range of about 3% to 10% final volume percent.
The reagent products of the present invention are useful for use in manual methods as well as automated methods in chemistry analysis and are particular useful for those
Field of the Invention This invention relates to stabilized control products useful in the clinical environment and in particular relates to stabilized coagulation control reagents and plasma-renin control reagents.
Background of the Invention This invention is related to a cop ending Canadian apply-cation Serial No. 455,085 entitled "A Stabilized Multi-parameter Control Product" (OARED) filed May 25, 1984 by the inventor hereof.
Hemostasis is a complicated life saving procedure involve in blood vessels plasma, platelets, vessel walls, neural and hum oral activity all for the purpose of stanching bleeding. Although there has been much research, especially within the last few decades, concerning the process of coagulation specifically and hemostasis in general, there is much that is not completely understood particularly with regard to the specific details concern-in the interaction of contributing mechanisms. For instance it is now known that there are no less than 35 compounds which take part in the formation of a firm blood clot. Ostensibly, blood coagulation is thus one of thy more complicated chemical processes occurring within the body In fact, there are so many separate components within the scheme of coagulation that the International Committee on nomenclature for Blood Coagulation formed by the International Hematology Congress has adopted a series of names which have now come into general acceptance. The most common of these names (with generally employed OARED
alternatives) and factor designations include the following:
Factor I Fibrinogen Factor II Prothrombin Factor III Thromboplastin Factor IV Calcium Factor V Proaccelerin, labile factor Factor VI (no longer used) Factor VII Serum prothrombin conversion accelerator (SPA), stable factor Factor VIII Antihemophilic factor (AH) Factor IX Christmas factor, plasma thromboplastin component (PTC) Factor X Stuart Factor, Stuart-Prower factor Factor XI Plasma thromboplastin antecedent (PTA) Factor XII Hegemony factor Factor XIII Fibrin stabilizing factor Profibrinolysin Plasminogen Fibrinolysin Plasm in Diagnostic determination of the presence and quantitation of various of the above factors is important as many clinically relevant diseases associated with abnormal clotting are associated therewith. For instance, the absence of Factor VIII is the most frequent and serious cause of genetically determined clotting defects such as hemophilia A which has been recognized for over 2,000 years. A deficiency in plasma concentration of Factor IX
is generally associated with the-genetically determined defect responsible for hemophilia B. hemorrhagic disease results from deficiencies in Factors X and XI whereas deficiencies in Factor XII induce longer clotting times I
but still permit eventual clotting. Diseases of the liver or Vitamin K deficiencies have also been associated with abnormal levels of clotting Factors II, VII, IX r and X as these are predominantly produced by the liver. Other causes of abnormal clotting may be related to reduced platelet counts, ~hrombocytopenia, often associated with abnormal levels of clotting Factors II, VII, IX, and X as these are predominantly produced by the liver. Other causes of abnormal clotting may be related to reduced platelet counts, thrombocytopenia, often associated with pernicious anemia, certain drug therapies, irradiation or increased peripheral destruction by antibodies.
The clinical value of testing for coagulation times is not limited to the detection of genetic or pathological disease states but is also useful in the regulation of anticoagulant therapy. Anticoagulants typically inhibit the coagulation mechanism such as by the heparin mediated inhibition of Factor X by Antithrombin III. Deficiencies in the clotting mechanisms due to congenital defects, pathological conditions, or anti coagulation therapy have been discussed generally in Chapter 7, "Coagulation and Hemostasis" by Robert D. Languidly, In Clinical Diagnosis, Davidson and Henry, 1979.
As commonly understood, coagulation may occur by two path-ways, the so called intrinsic pathway and extrinsic path-way. The former is generally triggered by the presence of a surface (thought to activate Factor XII) and, with the presence of phospholipids and calcium, through a number of steps eventually stimulates the formation of a stabilized fibrin clot. The partial thromboplastin time (PUT) test typically measures the intrinsic pathway wherein most congenital deficiencies occur. Consequently this type of test serves as an excellent presurgical coagulation screening test; however, since the reagent employed in OARED
us this test is typically a platelet substitute, the PUT test does not measure platelet activity.
The extrinsic pathway is generally triggered by injury to tissue and the resultant exudation of tissue thromboplas-tin acts on Factors VII and X which, following a series of steps, thereafter forms a fibrin clot. The extrinsic pathway is generally tested by the so called one-stage prothrombin time test to thereby detect most of the factors depressed by oral anticoagulant drugs for antic-agulant therapy control. The thrombin time test typically measures the quantity and reactivity of fibrinogen. This test is a rapid, semi quantitative test and is consequently the test of choice for intravascular clotting and fibrinolysis analysis. The above is presented in review in accompanying Table I as well as a booklet entitled "Concentric Concepts of Coagulation 1971, 1975" available from Ortho Diagnostics, Inc., Raritan, New Jersey.
OARED
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C.,1 I
I:: 9 Of:; Us S
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P' En I a Clue H O E-l r So O
H E I Jo H I
X p î a C
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Us Jo > H -1 1 I c I w pa x X Al I
Al I
Z Do H E-l Us 5.1 O
.,1 Q _ So O I -O
E-l E I P ;
With such attention being placed on the determination of a particular patient's coagulating ability and the presence or absence of specific factors, it is axiomatic that adequate controls must be available in order to ensure the proper operation of manual and automated methods designed to determine these factor levels and coagulation in general. Heretofore, such controls as were available have been typically characterized by relatively short shelf fifes both in the liquid and dry state due largely, in part, to the unstable nature of the ingredients or by their inability to withstand drying procedures such as lyophilization.
It us an object of the present invention to provide plasma-renin and coagulation control reagents having the appropriate levels of particular factors present therein in a stabilized format and capable of being lyophilized.
It is another object of the present invention to provide methods whereby coagulation control and plasma-renin control reagents may be stabilized It is another related object of the present invention to provide reagents in a stabilized format useful in manual and automated renal hypertensive testing and coagulation testing.
Brief Summary of the Invention In accordance with the objects and principles of the present invention, coagulation control reagents and plasma resin control reagents are provided which are substantially stabilized by the addition of plexiform stabilizing means. In the case of coagulation control reagents, anti coagulation means is added to substantially defibrinated plasma, whereby clotting components are held OARED
intact and residual fibrin formation is inhibited.
Additionally, plexiform stabilizing means, selected from the group consisting of sucrose, reducing monosaccharide sugars and reducing disaccharide sugars, is added so that S it is present in the final product in he range of about 3%-10% volume percent The preferred plexiform stabilizing means is selected from the group consisting of maltose, minutely, syllabus, glucose and lactose with the latter most being the most preferred. Additionally, and in accordance with the typo of coagulation control reagent desired, a specific clotting factor or factors may be removed such as Factor VIII and/or Factor I.
The plasma resin control reagent of the present invention comprises selected plasma to which has been added anti coagulation means whereby clotting components are held substantially intact, and plexiform stabilizing means similar to that described for the coagulation control reagent. The anti coagulation means is ideally chosen from eye group consisting of citrate, oxalate, ETA (ethylene Damon tetraacetic acid), heparin, and any combination of the foregoing.
The most preferred embodiments will have substantially all water removed by processes well known such as by lyophilization~
In another embodiment the present invention provides a method for producing a stabilized coagulation control reagent comprising the steps of: (a) providing whole blood; (b) removing substantially all cells from said whole blood to obtain plasma; (c) adding anti coagulation means whereby clotting components are held intact and fibrin formation is inhibited; (d) adjusting specific clotting Factor or Factors in accordance with the type of coagulation control reagent desired, (e) adding plexiform stabilizing means -pa-selected from the group consisting of sucrose, reducing monosaccharide sugars and reducing disaccharide sugars so that said plexiform stabilizing means is present in the range of about 3% to 10% final volume percent, In still another embodiment the present invention provides a stabilized plasma resin control resent useful for Anglo-tension testing comprising: (a) plasma having a known resin level; (b) anti coagulation means whereby clotting components are held intact and fibrin formation is substantially inhibited; and (c) plexiform stabilizing means selected from the group consisting of reducing monosaccharide sugars and reducing disaccharide sugars whereby said plexiform stabilizing means is present in the range of about 3% to 10% final volume percent.
The reagent products of the present invention are useful for use in manual methods as well as automated methods in chemistry analysis and are particular useful for those
2 tests involving coagulation and plasma resin activity.
These tests include, for instance, renal hypertension testing, actual coagulation testing, testing for angiotension I Andre II, and resin activity testing '7~5i which is typically run only on fresh human plasma. As may be expected, particularly with the latter most test, adequate control reagents are difficult to obtain particularly in a suitably stabilized format.
s Such a stabilized format is particularly desirous in the clinical environment in order to minimize cost expenditures and to provide increased comparative integrity of the data on a daily testing basis.
Ideally, the control reagent materials will simulate a patient's sample as closely as possible except, to the extent the controls may be artificially altered in order to provide factor deficient controls to levels which may be reasonably expected with particular disease states.
For instance, a patient suffering a cardiac infarction may be expected to have increased levels of resin. Resin is known to act on plasma proteins thereby forming angiotensin, in turn, responsible for an increase in blood pressure occasioned by constricted blood vessels. The resin, a proteolytic enzyme believed to be produced in the efferent golmerular arterioles, reacts with hypertensinogen to product angiotensin II. The latter is a decapeptide hormone which, in addition to influencing US blood vessel constriction, also alters aldosterone secretion by the adrenal cortex. Angiotensin is also known as hypertension. Aldosterone is a steroid hormone which function chiefly in regulating sodium and potassium metabolism and thereby it also plays an important part in cardiac function.
Accordingly, and in order to ensure accurate testing procedures, a plasma resin control reagent is a virtual necessity, and is particularly desirable if made in a stable format. In accordance with the objects of the present invention, such a control reagent is now made OARED
7~5 g hereby possible. It comprises plasma, ideally from human origin, preferably selected for its high resin levels.
Additionally, an anticoagulant is added whereby the clotting components may be held intact and fibrin formation inhibited. Such an anticoagulant may ye selected from the group consisting of Satyr oxalate, ETA, heparin or a combination thereof.
The desired stability is obtained by adding plexiform stabilizing means selected from the group consisting of reducing monosaccharide sugars and reducing disaccharide sugars Preferred embodiments employ maltose, minutely, syllabus, lactose or glucose. The plexiform stabilizing means is added so that the final percent concentration is preferably in the range of about 3%-10%. As more fully described in the OARED application Serial Noah referenced earlier, the plexiform stabilizing means is thought to provide a three dimensional crystalline" structure whereby the proteins, analyzes, and constituents of the control reagents are held in stabilized juxtaposition. It is further believed by the inventor hereof, although he does not wish such belief to be a limitation hereof, that the plexiform stabilizing means serves to hold the individual constituents in a substantially rigid manner thereby reducing the amount of degradation or lability which may be associated or occasioned by the physical movement of the analyzes.
Further, the plexiform stabilizing means serves to assist in the sublimation of water during water removal processes such as by lyophilization. The actual mechanism by which this is accomplished is unknown, however, it has been observed, particularly with respect to the clinical chemistry control reagents of the OARED application, that reconstitution of the reagent after lyophilization is accomplished with greater speed and with increased vial-to-vial reproducibility These advantages may also be expected following lyophilization of the control reagents of the present invention.
Similarly, the coagulation control reagents are ideally produced from plasma having added thereto an anticoagulant such as that described above, whereby the clotting components are held intact and residual fibrin formation is substantially inhibited. In addition to supplementation with the above-mentioned plexiform stabilizing means, the coagulation control reagent may be adjusted to remove specific clotting factor or factors in accordance with the type of coagulation control reagent desired. For instance, should a reagent be desired which is sub Stan-tidally deficient in Factor I ~fibrinogen~ or Factor VIII
(antihaemophelic factor), then those components may be removed such as by ethanol extraction or chilling, respectively, or by other methods for other Factors as is well-known in the art.
As with the plasma resin control reagent, the plexiform stabilizing means acts to provide a three dimensional structure to greatly increase stability, speed of reconstitution following lyophilization and superior vial-to-vial reproducibility. Further, and as is more fully explained in the OARED application SUN Thea speed with which lyophilization is accomplished may actually be increased despite the lowering of the freezing point This is thought to be generally due to the of imitation of the eutectic point plateau. Additionally, and for reasons not yet fully explained, the reconstituted materials typically exhibit greater optical clarity OARED
This is probably due, at least in part, to the decreased denaturation of proteins, particularly a problem during lyophilization.
Still other advantages are gained by the instant invention and include the observation that the defibrinated plasma appears more completely defibrinated with the addition of the plexiform stabilizing means than defibrinated plasma not so augmented. The observed, more consistent gel-like clot may thus be a natural progression therefrom and may also be indicative of the enhanced overall stability of other inherent clotting factors present in the plasma.
The most preferred reagents of the present invention will also have substantially all water removed by processes well-known in the art. Such processes include, without limitation, lyophilization and the like, and serve to afford even greater shelf lives. Ideally, the resultant dry product is stored at approximately 2- 8C and reconstituted just prior to use.
Stability as used herein shall mean later measured activity which is at least 90~ of the original measured activity.
As may be readily appreciated by those skilled in the art, numerous insubstantial alterations or substitutions of the aforementioned components may be made without departure from the spirit or scope of the present invention.
OARED
These tests include, for instance, renal hypertension testing, actual coagulation testing, testing for angiotension I Andre II, and resin activity testing '7~5i which is typically run only on fresh human plasma. As may be expected, particularly with the latter most test, adequate control reagents are difficult to obtain particularly in a suitably stabilized format.
s Such a stabilized format is particularly desirous in the clinical environment in order to minimize cost expenditures and to provide increased comparative integrity of the data on a daily testing basis.
Ideally, the control reagent materials will simulate a patient's sample as closely as possible except, to the extent the controls may be artificially altered in order to provide factor deficient controls to levels which may be reasonably expected with particular disease states.
For instance, a patient suffering a cardiac infarction may be expected to have increased levels of resin. Resin is known to act on plasma proteins thereby forming angiotensin, in turn, responsible for an increase in blood pressure occasioned by constricted blood vessels. The resin, a proteolytic enzyme believed to be produced in the efferent golmerular arterioles, reacts with hypertensinogen to product angiotensin II. The latter is a decapeptide hormone which, in addition to influencing US blood vessel constriction, also alters aldosterone secretion by the adrenal cortex. Angiotensin is also known as hypertension. Aldosterone is a steroid hormone which function chiefly in regulating sodium and potassium metabolism and thereby it also plays an important part in cardiac function.
Accordingly, and in order to ensure accurate testing procedures, a plasma resin control reagent is a virtual necessity, and is particularly desirable if made in a stable format. In accordance with the objects of the present invention, such a control reagent is now made OARED
7~5 g hereby possible. It comprises plasma, ideally from human origin, preferably selected for its high resin levels.
Additionally, an anticoagulant is added whereby the clotting components may be held intact and fibrin formation inhibited. Such an anticoagulant may ye selected from the group consisting of Satyr oxalate, ETA, heparin or a combination thereof.
The desired stability is obtained by adding plexiform stabilizing means selected from the group consisting of reducing monosaccharide sugars and reducing disaccharide sugars Preferred embodiments employ maltose, minutely, syllabus, lactose or glucose. The plexiform stabilizing means is added so that the final percent concentration is preferably in the range of about 3%-10%. As more fully described in the OARED application Serial Noah referenced earlier, the plexiform stabilizing means is thought to provide a three dimensional crystalline" structure whereby the proteins, analyzes, and constituents of the control reagents are held in stabilized juxtaposition. It is further believed by the inventor hereof, although he does not wish such belief to be a limitation hereof, that the plexiform stabilizing means serves to hold the individual constituents in a substantially rigid manner thereby reducing the amount of degradation or lability which may be associated or occasioned by the physical movement of the analyzes.
Further, the plexiform stabilizing means serves to assist in the sublimation of water during water removal processes such as by lyophilization. The actual mechanism by which this is accomplished is unknown, however, it has been observed, particularly with respect to the clinical chemistry control reagents of the OARED application, that reconstitution of the reagent after lyophilization is accomplished with greater speed and with increased vial-to-vial reproducibility These advantages may also be expected following lyophilization of the control reagents of the present invention.
Similarly, the coagulation control reagents are ideally produced from plasma having added thereto an anticoagulant such as that described above, whereby the clotting components are held intact and residual fibrin formation is substantially inhibited. In addition to supplementation with the above-mentioned plexiform stabilizing means, the coagulation control reagent may be adjusted to remove specific clotting factor or factors in accordance with the type of coagulation control reagent desired. For instance, should a reagent be desired which is sub Stan-tidally deficient in Factor I ~fibrinogen~ or Factor VIII
(antihaemophelic factor), then those components may be removed such as by ethanol extraction or chilling, respectively, or by other methods for other Factors as is well-known in the art.
As with the plasma resin control reagent, the plexiform stabilizing means acts to provide a three dimensional structure to greatly increase stability, speed of reconstitution following lyophilization and superior vial-to-vial reproducibility. Further, and as is more fully explained in the OARED application SUN Thea speed with which lyophilization is accomplished may actually be increased despite the lowering of the freezing point This is thought to be generally due to the of imitation of the eutectic point plateau. Additionally, and for reasons not yet fully explained, the reconstituted materials typically exhibit greater optical clarity OARED
This is probably due, at least in part, to the decreased denaturation of proteins, particularly a problem during lyophilization.
Still other advantages are gained by the instant invention and include the observation that the defibrinated plasma appears more completely defibrinated with the addition of the plexiform stabilizing means than defibrinated plasma not so augmented. The observed, more consistent gel-like clot may thus be a natural progression therefrom and may also be indicative of the enhanced overall stability of other inherent clotting factors present in the plasma.
The most preferred reagents of the present invention will also have substantially all water removed by processes well-known in the art. Such processes include, without limitation, lyophilization and the like, and serve to afford even greater shelf lives. Ideally, the resultant dry product is stored at approximately 2- 8C and reconstituted just prior to use.
Stability as used herein shall mean later measured activity which is at least 90~ of the original measured activity.
As may be readily appreciated by those skilled in the art, numerous insubstantial alterations or substitutions of the aforementioned components may be made without departure from the spirit or scope of the present invention.
OARED
Claims (48)
1. A stabilized coagulation control reagent comprising:
a) plasma;
b) anticoagulant means whereby clotting components are held intact and fibrin formation is inhibited;
c) plexiform stabilizing means selected from the group consisting of sucrose, reducing monosaccharide sugars and reducing disaccharide sugars present in the range of about 3%-10% final volume percent whereby the integrity of clotting components is augmented; and wherein d) said plasma may be manipulated to adjust specific clotting Factor or Factors in accordance with the type of coagulation control reagent desired.
a) plasma;
b) anticoagulant means whereby clotting components are held intact and fibrin formation is inhibited;
c) plexiform stabilizing means selected from the group consisting of sucrose, reducing monosaccharide sugars and reducing disaccharide sugars present in the range of about 3%-10% final volume percent whereby the integrity of clotting components is augmented; and wherein d) said plasma may be manipulated to adjust specific clotting Factor or Factors in accordance with the type of coagulation control reagent desired.
2. The reagent as provided in Claim 1 wherein the plexiform stabilizing means is selected from the group consisting of sucrose, lactose, mannitol, cellobiose and maltose.
3. The reagent as provided in Claim 2 wherein the plasma is human plasma.
4. The reagent as provided in Claim 3 wherein the anticoagulant is selected from the group consisting of citrate, oxalate, EDTA, heparin, and combinations thereof.
5. The reagent as provided in Claim 4 wherein the clotting factors removed are selected from the group consisting of factor 1, factor 8 and a combination thereof.
6. The reagent as provided in Claim 4 wherein the plexiform stabilizing means is lactose.
7. The reagent as provided in Claim 4 wherein the plexiform stabilizing means is sucrose.
8. The reagent as provided in Claim 4 wherein the plexiform stabilizing means is cellobiose.
9. The reagent as provided in Claim 4 having substantially all water removed.
10. The reagent as provided in Claim 6 having substantially all water removed.
11. The reagent as provided in Claim 7 having substantially all water removed.
12. The reagent as provided in Claim 8 having substantially all water removed
13. A method for producing a stabilized coagulation control reagent comprising the steps of:
a) providing whole blood;
b) removing substantially all cells from said whole blood to obtain plasma;
c) adding anticoagulation means whereby clotting components are held intact and fibrin formation is inhibited;
d) adjusting specific clotting Factor or Factors in accordance with the type of coagulation control reagent desired;
e) adding plexiform stabilizing means selected from the group consisting of sucrose, reducing monosaccharide sugars and reducing disaccharide sugars so that said plexiform stabilizing means is present in the range of about 3% to 10% final volume percent.
a) providing whole blood;
b) removing substantially all cells from said whole blood to obtain plasma;
c) adding anticoagulation means whereby clotting components are held intact and fibrin formation is inhibited;
d) adjusting specific clotting Factor or Factors in accordance with the type of coagulation control reagent desired;
e) adding plexiform stabilizing means selected from the group consisting of sucrose, reducing monosaccharide sugars and reducing disaccharide sugars so that said plexiform stabilizing means is present in the range of about 3% to 10% final volume percent.
14. The method as provided in Claim 13 wherein the plexiform stabilizing means is selected from the group consisting of sucrose, lactose, mannitol, cellobiose and maltose.
15. The method as provided in Claim 13 wherein the plexiform stabilizing means is lactose.
16. The method as provided in Claim 14 wherein said anticoagulation means is selected from the group consisting of citrate, oxalate, EDTA, heparin and combinations thereof.
17. The method as provided in Claim 15 wherein said anticoagulation means is selected from the group consisting of citrate, oxalate, EDTA, heparin and combinations thereof.
18. The method as provided in Claim 16 wherein the Factor removing step comprises a step selected from the group consisting of removing Factor VIII by chilling, removing Factor I by ethanol extraction, and combinations thereof.
19. The method as provided in Claim 17 wherein the removing step comprises a step selected from the group consisting of removing Factor VIII by chilling, removing Factor I by ethenol extraction and combinations thereof.
20. The method as provided in Claim 13 further comprising the step of removing substantially all water.
21. The method as provided in Claim 18 further comprising the step of removing substantially all water.
22. The method as provided in Claim 19 further comprising the step of removing substantially all water.
23. The method as provided in Claim 20 wherein the water removing step is accomplished by lyophilizing.
24. The method as provided in Claim 21 wherein the water removing step is accomplished by lyophilization.
25. The method as provided in Claim 22 wherein the water removing step is accomplished by lyophilization.
26. A stabilized plasma renin control reagent useful for angiotensin testing comprising:
a) plasma having a known renin level ;
b) anticoagulation means whereby clotting components are held intact and fibrin formation is substantially inhibited; and c) plexiform stabilizing means selected from the group consisting of reducing monosaccharide sugars and reducing disaccharide sugars whereby said plexiform stabilizing means is present in the range of about 3% to 10% final volume percent.
a) plasma having a known renin level ;
b) anticoagulation means whereby clotting components are held intact and fibrin formation is substantially inhibited; and c) plexiform stabilizing means selected from the group consisting of reducing monosaccharide sugars and reducing disaccharide sugars whereby said plexiform stabilizing means is present in the range of about 3% to 10% final volume percent.
27. The reagent as provided in Claim 26 wherein the plexiform stabilizing means is selected from the group consisting of maltose, mannitol, cellobiose, lactose and glucose.
28. The reagent as provided in Claim 26 wherein the plexiform stabilizng means is lactose.
29. The reagent as provided in Claim 27 wherein the plasma is human plasma.
30. The reagent as provided in Claim 28 wherein the plasma is human plasma.
31. The reagent as provided in Claim 29 wherein said anticoagulation means is selected from the group consisting of citrate, oxalate, EDTA, heparin, and combinations thereof.
32. The reagent as provided in Claim 30 wherein said anticoagulation means is selected from the group consisting of citrate, oxalate, EDTA, heparin, and combinations thereof.
33. The reagent as provided in Claim 26 having substantially all water removed.
34. The reagent as provided in Claim 27 having substantially all water removed.
35. The reagent as provided in Claim 28 having substantially all water removed.
36. The reagent as provided in Claim 29 having substantially all water removed.
37. The reagent as provided in Claim 30 having substantially all water removed.
38. The reagent as provided in Claim 31 having substantially all water removed.
39. A method for producing a stabilized plasma renin control reagent suitable for angiotensin testing comprising the steps of:
a) selecting a plasma having a known renin activity;
b) adding anticoagulation means whereby the clotting components are held intact and fibrin formation is substantially inhibited; and c) adding plexiform stabilizing means selected from the group consisting of reducing monosaccharide sugars and reducing disaccharide sugars so that said plexiform stabilizing means is present in a final volume percent in the range of about 3% to 10%.
a) selecting a plasma having a known renin activity;
b) adding anticoagulation means whereby the clotting components are held intact and fibrin formation is substantially inhibited; and c) adding plexiform stabilizing means selected from the group consisting of reducing monosaccharide sugars and reducing disaccharide sugars so that said plexiform stabilizing means is present in a final volume percent in the range of about 3% to 10%.
40. The method as provided in Claim 39 wherein said plexiform stabilizing means is selected from the group consisting of maltose, mannitol, cellobiose, lactose and glucose.
41. The method as provided in Claim 39 wherein said plexiform stabilizing means is lactose.
42. The method as provided in Claim 40 wherein the plasma is human plasma and the anticoagulation means is selected from the group consisting of citrate, oxalate, EDTA, heparin and combinations thereof.
43. The method as provided in Claim 41 wherein the plasma is human plasma and the anticoagulation means is selected from the group consisting of citrate, oxalate, EDTA, heparin and combinations thereof.
44. The method as provided in Claim 39 further comprising the step of removing substantially all water.
45. The method as provided in Claim 40 further comprising the step of removing substantially all water.
46. The method as provided in Claim 41 further comprising the step of removing substantially all water.
47. The method as provided in Claim 42 further comprising the step of removing substantially all water.
48. The method as provided in Claim 43 further comprising the step of removing substantially all water.
Applications Claiming Priority (2)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| US50121683A | 1983-06-06 | 1983-06-06 | |
| US501,216 | 1983-06-06 |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CA1226795A true CA1226795A (en) | 1987-09-15 |
Family
ID=23992589
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CA000455086A Expired CA1226795A (en) | 1983-06-06 | 1984-05-25 | Stabilized coagulation control products |
Country Status (6)
| Country | Link |
|---|---|
| JP (2) | JPS6018762A (en) |
| AU (1) | AU584859B2 (en) |
| CA (1) | CA1226795A (en) |
| DK (1) | DK275384A (en) |
| IL (1) | IL72032A0 (en) |
| NO (1) | NO842260L (en) |
Families Citing this family (8)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US4663295A (en) * | 1983-06-29 | 1987-05-05 | Ciba Corning Diagnostics Corp. | Estrogen-progesterone control reagents and methods for making same |
| CA1226795A (en) * | 1983-06-06 | 1987-09-15 | Michael K. Hoskins | Stabilized coagulation control products |
| CA1226794A (en) * | 1983-06-06 | 1987-09-15 | Michael K. Hoskins | Stabilized multiparameter control product |
| CA1230300A (en) * | 1983-06-06 | 1987-12-15 | Michael K. Hoskins | Stabilized isoenzyme control product |
| JPH0695092B2 (en) * | 1989-10-30 | 1994-11-24 | 隆己 永田 | Anticoagulant for blood cell counting |
| JP4592422B2 (en) * | 2002-11-18 | 2010-12-01 | デンカ生研株式会社 | Immunoassay to prevent divergence of serum and plasma measurements |
| US7588942B2 (en) * | 2006-08-04 | 2009-09-15 | Bio-Rad Laboratories, Inc. | Standard/reference/control for blood coagulation testing |
| EP2405275B1 (en) * | 2009-03-05 | 2018-10-10 | BML, Inc. | Method for measuring glycated albumin contained in serum or plasma using mannitol as stabiliser for a control serum or plasma. |
Family Cites Families (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| DE2461969A1 (en) * | 1974-12-31 | 1976-07-08 | Behringwerke Ag | STABLE BLOOD PLASMA, THE PROCESS FOR PRODUCING IT AND ITS USE AS A COMPARATIVE PLASMA IN COOLAGE EXAMINATIONS |
| JPS5757661A (en) * | 1980-09-25 | 1982-04-06 | Daicel Ltd | Film for separately packing sliced cheese |
| CA1226795A (en) * | 1983-06-06 | 1987-09-15 | Michael K. Hoskins | Stabilized coagulation control products |
| CA1226794A (en) * | 1983-06-06 | 1987-09-15 | Michael K. Hoskins | Stabilized multiparameter control product |
| CA1230300A (en) * | 1983-06-06 | 1987-12-15 | Michael K. Hoskins | Stabilized isoenzyme control product |
| JPH0511264A (en) * | 1991-07-05 | 1993-01-19 | Hitachi Ltd | Liquid crystal display device |
-
1984
- 1984-05-25 CA CA000455086A patent/CA1226795A/en not_active Expired
- 1984-06-04 DK DK275384A patent/DK275384A/en not_active Application Discontinuation
- 1984-06-04 JP JP11322684A patent/JPS6018762A/en active Granted
- 1984-06-05 AU AU29086/84A patent/AU584859B2/en not_active Ceased
- 1984-06-05 NO NO842260A patent/NO842260L/en unknown
- 1984-06-05 IL IL72032A patent/IL72032A0/en unknown
-
1991
- 1991-12-10 JP JP34986791A patent/JPH0560753A/en active Pending
Also Published As
| Publication number | Publication date |
|---|---|
| DK275384D0 (en) | 1984-06-04 |
| AU2908684A (en) | 1984-12-13 |
| JPS6018762A (en) | 1985-01-30 |
| NO842260L (en) | 1984-12-07 |
| JPH0560753A (en) | 1993-03-12 |
| JPH0511264B2 (en) | 1993-02-15 |
| IL72032A0 (en) | 1984-10-31 |
| DK275384A (en) | 1984-12-07 |
| AU584859B2 (en) | 1989-06-08 |
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