Quality control formulations for blood coagulation analyzers

Abstract

Quality control formulations used to test the effectiveness of viscoelastic analysis reagents that do not require blood components derived from human donors, and methods for preparing such formulations, are described.

Description

[Technical Field]

[0001] Cross-reference to related applications This application claims priority to U.S. Provisional Application No. 63 / 168,838, filed on 31 March 2021, all disclosures thereof are incorporated herein by reference.

[0002] The present invention relates to quality control formulations used for testing the effectiveness of viscoelastic analytical reagents and methods for preparing such formulations. [Background technology]

[0003] Viscoelastic analysis is crucial for evaluating patient blood samples in a wide variety of medical settings. Therefore, it is essential that the reagents used in viscoelastic analysis provide accurate results, especially when used in hospitals and clinical laboratories. To ensure these reagents deliver accurate and reliable results, it is important to conduct quality control testing on the reagents before they reach the end user.

[0004] Current formulations for quality control testing of viscoelastic analysis reagents require human donors, human blood, human plasma, or components derived from human blood, such as human red blood cells. The use of quality control formulations containing blood components derived from human donors is relatively costly and poses an infection risk to the donor.

[0005] Therefore, there is a need for reliable quality control formulations for testing viscoelastic analysis reagents that do not contain blood components derived from human donors. [Overview of the project]

[0006] According to one embodiment of the present invention, a method is provided for preparing a formulation for testing the effectiveness of at least one viscoelastic analysis reagent, the formulation having a target citrated bovine plasma concentration, a target human tissue factor concentration, and a target heparin concentration. The method comprises a) determining a target concentration of citrated bovine plasma for the formulation by preparing a citrated bovine plasma dilution series having at least two members; measuring a first viscoelastic property for each member of the citrated bovine plasma dilution series to obtain a first value for each member of the citrated bovine plasma dilution series, the first viscoelastic property having a target value; calculating a first linear regression from the first value for each member of the citrated bovine plasma dilution series; and extrapolating a predicted citrated bovine plasma concentration corresponding to the first viscoelastic property target value from the first linear regression, the predicted citrated bovine plasma concentration being the target citrated bovine plasma concentration. This method further includes b) determining a target concentration of human tissue factor for a formulation by preparing a human tissue factor dilution series, the human tissue factor dilution series having at least two members; measuring a second viscoelastic property for each member of the human tissue factor dilution series to obtain a second value for each member of the human tissue factor dilution series, the second viscoelastic property having a target value; calculating a second linear regression from the second value for each member of the human tissue factor dilution series; and extrapolating a predicted human tissue factor concentration corresponding to the second viscoelastic property target value from the second linear regression, the predicted human tissue factor concentration being the target human tissue factor concentration. This method also includes c) determining a target concentration of heparin for a formulation by preparing a heparin dilution series, the heparin dilution series having at least two members, measuring a third viscoelastic property for each member of the heparin dilution series to obtain a third value for each member of the heparin dilution series, the third viscoelastic property having a target value, calculating a third linear regression from the third value for each member of the heparin dilution series; and extrapolating a predicted heparin concentration corresponding to the third viscoelastic property target value from the third linear regression, the predicted heparin concentration being the target heparin concentration.This method further includes preparing a formulation by combining citrate-bovine plasma, human tissue factor, and heparin, such that the formulation contains citrate-bovine plasma at a target citrate-bovine plasma concentration, human tissue factor at a target human tissue factor concentration, and heparin at a target heparin concentration.

[0007] In some embodiments, steps b) and c) may be performed simultaneously.

[0008] In some embodiments, in step (a), each member of the citrated bovine plasma dilution series has a citrated bovine plasma concentration, the concentration of citrated bovine plasma in any one member of the citrated bovine plasma dilution series is different from the concentration of citrated bovine plasma in any other member of the citrated bovine plasma dilution series, and each member of the citrated bovine plasma dilution series has a human tissue factor concentration, the human tissue factor concentrations of each member of the citrated bovine plasma dilution series are the same. In procedure (b), each member of the human tissue factor dilution series has a human tissue factor concentration, the concentration of human tissue factor in any one member of the human tissue factor dilution series is different from the concentration of human tissue factor in any other member of the human tissue factor dilution series, each member of the human tissue factor dilution series has a citrated bovine plasma concentration, the citrated bovine plasma concentration of each member of the human tissue factor dilution series is the target citrated bovine plasma concentration, and each member of the human tissue factor dilution series has a heparin concentration, the heparin concentrations of each member of the human tissue factor dilution series are the same. In procedure (c), each member of the heparin dilution series has a heparin concentration, the heparin concentration in any one member of the heparin dilution series is different from the heparin concentration in any other member of the heparin dilution series, each member of the heparin dilution series has a citrated bovine plasma concentration, the citrated bovine plasma concentration of each member of the heparin dilution series is the target citrated bovine plasma concentration, and each member of the heparin dilution series has a human tissue factor concentration, the human tissue factor concentration of each member of the heparin dilution series is the target human tissue factor concentration.

[0009] In other embodiments, in step (a), each member of the citrate-bovine plasma dilution series has a citrate-bovine plasma concentration, the concentration of citrate-bovine plasma in any one member of the citrate-bovine plasma dilution series is different from the concentration of citrate-bovine plasma in any other member of the citrate-bovine plasma dilution series, and each member of the citrate-bovine plasma dilution series has a human tissue factor concentration, the human tissue factor concentrations of each member of the citrate-bovine plasma dilution series are the same. In procedure (c), each component of the heparin dilution series has a heparin concentration, the heparin concentration in any one component of the heparin dilution series is different from the heparin concentration in any other component of the heparin dilution series, each component of the heparin dilution series has a citrated bovine plasma concentration, the citrated bovine plasma concentration of each component of the heparin dilution series is the target citrated bovine plasma concentration, and each component of the heparin dilution series has a human tissue factor concentration, the human tissue factor concentrations of each component of the heparin dilution series are the same. In procedure (b), each member of the human tissue factor dilution series has a human tissue factor concentration, the concentration of human tissue factor in any one member of the human tissue factor dilution series is different from the concentration of human tissue factor in any other member of the human tissue factor dilution series, each member of the human tissue factor dilution series has a citrated bovine plasma concentration, the citrated bovine plasma concentration of each member of the human tissue factor dilution series is the target citrated bovine plasma concentration, and each member of the human tissue factor dilution series has a heparin concentration, the heparin concentration of each member of the human tissue factor dilution series is the target heparin concentration.

[0010] According to another embodiment of the present invention, a method is provided for preparing a formulation for testing the effectiveness of at least one viscoelastic analysis reagent, the formulation having a target concentration of citrated bovine plasma, a target concentration of human tissue factor, and a target concentration of heparin. The method involves a) measuring a first viscoelastic property of a citrated bovine plasma dilution to obtain a first value for the citrated bovine plasma dilution, the first viscoelastic property having a target value, and determining a predicted citrated bovine plasma concentration corresponding to the first viscoelastic property target value based on the first value, and determining the target concentration of citrated bovine plasma for the formulation, wherein the predicted citrated bovine plasma concentration is the target citrated bovine plasma concentration; and b) measuring a second viscoelastic property of a human tissue factor dilution to obtain a second value for the human tissue factor dilution, the second viscoelastic property having a target value, and determining a predicted human tissue factor concentration corresponding to the second viscoelastic property target value based on the second value, wherein the predicted human tissue factor concentration is the target human tissue The method includes: a) determining the target concentration of human tissue factor for the formulation by determining the factor concentration; c) measuring a third viscoelastic property of the heparin dilution to obtain a third value for the heparin dilution, wherein the third viscoelastic property has a target value, and determining the predicted heparin concentration based on the third value corresponding to the target value of the third viscoelastic property, wherein the predicted heparin concentration is the target heparin concentration, thereby determining the target concentration of heparin for the formulation; and d) preparing the formulation by combining citrated bovine plasma, human tissue factor, and heparin so that the formulation contains citrated bovine plasma at a target citrated bovine plasma concentration, human tissue factor at a target human tissue factor concentration, and heparin at a target heparin concentration.

[0011] In some embodiments, steps b) and c) may be performed simultaneously.

[0012] According to another embodiment of the present invention, a method is provided for preparing a formulation for testing the effectiveness of at least one viscoelastic analytical reagent, the formulation having a target citrate-added bovine plasma concentration, a target human tissue factor concentration, and a target heparin concentration. This method includes: a) preparing a citrate-added bovine plasma dilution series having at least two components, measuring a first viscoelastic property for each component of the citrate-added bovine plasma dilution series to obtain a first value for each component of the citrate-added bovine plasma dilution series, the first viscoelastic property having a target value, calculating a first linear regression from the first value for each component of the citrate-added bovine plasma dilution series, and extrapolating a predicted citrate-added bovine plasma concentration corresponding to the first viscoelastic property target value from the first linear regression, thereby determining the target citrate-added bovine plasma concentration for the formulation by having this predicted citrate-added bovine plasma concentration be the target citrate-added bovine plasma concentration; and b) determining the target human tissue factor concentration and target heparin concentration for the formulation by preparing a set of human tissue factor dilutions, the set of human tissue factor dilutions having at least two heparin dilution series, each heparin dilution series having at least The method also includes having two members, where each member of each heparin dilution series has a human tissue factor concentration and a heparin concentration, and a second viscoelastic property is measured for each member of each heparin dilution series to obtain a second value for each member of each heparin dilution series, the second viscoelastic property having a target value, and a selected member from the human tissue factor dilution set is identified, having a measured second viscoelastic property that is closer to the target value of the second viscoelastic property than the other members of the human tissue factor dilution set, the human tissue factor concentration of the selected member being the target human tissue factor concentration, and the heparin concentration of the selected member being the target heparin concentration; and c) preparing a formulation by combining citrated bovine plasma, human tissue factor, and heparin together, such that the formulation contains citrated bovine plasma at a target citrated bovine plasma concentration, human tissue factor at a target human tissue factor concentration, and heparin at a target heparin concentration.

[0013] In some embodiments, in step (a), each member of the citrated bovine plasma dilution series has a citrated bovine plasma concentration, the concentration of citrated bovine plasma in any one member of the citrated bovine plasma dilution series is different from the concentration of citrated bovine plasma in any other member of the citrated bovine plasma dilution series, and each member of the citrated bovine plasma dilution series has a human tissue factor concentration, the human tissue factor concentrations of each member of the citrated bovine plasma dilution series are the same. In procedure (b), each component of each heparin dilution series has a citrated bovine plasma concentration, the citrated bovine plasma concentration of each component of each heparin dilution series is the target citrated bovine plasma concentration, the concentration of human tissue factor in any one heparin dilution series is different from the concentration of human tissue factor in any other heparin dilution series, the concentrations of human tissue factor in all components of any one heparin dilution series are the same, and the concentration of heparin in any one component of any one heparin dilution series is different from the concentration of heparin in any other component of the same heparin dilution series.

[0014] The first, second, and / or third viscoelastic properties may be measured using a device for viscoelastic analysis. The device for viscoelastic analysis may be a microfluidic cartridge.

[0015] The first viscoelastic property may be the MA value, and the target value for the first viscoelastic property may be approximately 63. The second viscoelastic property may be the R value, and the target value for the second viscoelastic property may be approximately 6. The third viscoelastic property may be the R value, and the target value for the third viscoelastic property may be approximately 6.

[0016] According to one embodiment of the present invention, the formulation is for testing the effectiveness of at least one viscoelastic analysis reagent, the formulation comprising 60-90% citrated bovine plasma, 0.05-0.20% human tissue factor, and 0.05-0.30 U / ml heparin.

[0017] In some embodiments, the formulation may include a stabilizer. The stabilizer may be glycine or a HEPES buffer. The formulation may include a preservative. The preservative may be sodium azide. The pH of the formulation may be from about 7.3 to about 7.9. The total protein content of the formulation may exceed about 6.4 g / dL.

Brief Description of the Drawings

[0018] The features of the above embodiments will be more readily understood by referring to the following detailed description with reference to the accompanying drawings. In the accompanying drawings:

[0019] FIG. 1 shows the test results of a TEG global blood coagulation cartridge obtained using human donor blood under simulated deteriorated CaCl2 conditions according to an embodiment of the present invention.

[0020] FIG. 2 shows the test results of a TEG global blood coagulation cartridge obtained using the quality control formulation described in the present application under simulated deteriorated CaCl2 conditions according to an embodiment of the present invention.

[0021] FIG. 3 shows the test results of a TEG global blood coagulation cartridge obtained using the quality control formulation described in the present application under simulated deteriorated heparinase conditions according to an embodiment of the present invention.

Modes for Carrying Out the Invention

[0022] Definitions. As used in this detailed description and the appended claims, unless the context indicates otherwise, the following terms have the meanings set forth herein:

[0023] With respect to numerical values, "about" means ± 10% of the indicated numerical value.

[0024] "Viscoelastic analysis" means any analytical method for measuring the properties of elastic solids (e.g., fibrin solids) and fluids. In other words, viscoelastic analysis makes it possible to study the properties of viscous fluids such as blood, plasma, or blood samples. In some embodiments, the viscoelastic analysis is performed under conditions that mimic the in vivo state that results in blood clotting. For example, these conditions may include a temperature that mimics body temperature (e.g., a temperature of 37°C). These conditions may also include the formation and dissolution of blood clots at a flow rate that mimics the flow rate found in blood vessels. Viscoelastic analysis includes, but is not limited to, viscoelastic assays and coagulation assays, such as those performed using thromboelastography (TEG), thromboelastometry (TEM), and microfluidic cartridges, such as the TEG global blood coagulation assay cartridge suitable for use with the TEG6s blood coagulation analysis system (Hemogenetics, Inc., Boston, Massachusetts).

[0025] "Viscoelastic properties", "parameters", "coagulation measurements", and like terms may mean measurements obtained through viscoelastic analysis, such as measurement of blood clot formation. This measurement can be made at any point during blood clot formation, including, but not limited to, the point at which fibrin is first formed or the point at which the blood clot reaches a predetermined level of strength.

[0026] One non-limiting example of a viscoelastic property or parameter is the initial clot formation time ("R"), which is the time it takes for fibrin to first form. Another non-limiting example of a viscoelastic property or parameter is "MA" (maximum amplitude in mm), which is a direct function of the maximum dynamic properties of fibrin-platelet binding and represents the final strength of the fibrin-platelet clot. If the platelet function of the blood component being tested is impaired, the MA value represents the strength of the clot based solely on fibrin. Viscoelastic properties can be measured for blood components obtained directly from the subject, for blood components treated with coagulation activators such as kaolin, or for blood components treated with anticoagulants such as citrate, supplemented as appropriate by calcium addition.

[0027] The term "viscoelastic analysis device" and its synonyms refer to an instrument or device that can be used to measure the viscoelastic properties of elastic solids and fluids, such as a device used to measure the ability of blood or blood components to form a blood clot. Microfluidic cartridges, such as the TEG Global Blood Coagulation Assay Cartridge suitable for use with the TEG6s Blood Coagulation Analysis System (Hemonetics, Boston, Massachusetts), are non-limiting examples of devices for viscoelastic analysis.

[0028] The term "viscoelastic analysis reagent" and its synonyms refer to any compound other than blood or plasma test samples used to perform viscoelastic analysis on a test sample. Non-limiting examples of viscoelastic analysis reagents include calcium chloride (CaCl2) and biomolecules such as heparinase.

[0029] As used in this application, "target value" and its synonyms mean a certain value (or range of values) of a viscoelastic property or parameter for a particular coagulation assay, which is a preferred value when the particular assay is performed on blood components from a healthy subject using a viscoelastic analytical reagent that has not deteriorated to a detectable degree or lost its effectiveness.

[0030] As used in this application, "target concentration" and its synonyms refer to a certain concentration of a component in a quality control formulation, which is the concentration at which, when present in the quality control formulation at that concentration, a specific assay is determined or predicted to be achieved when performed with a viscoelastic analysis reagent that has not deteriorated to a detectable degree or lost its effectiveness.

[0031] Quality control of reagents used for the viscoelastic analysis of human blood, including quality control of devices containing such reagents, is crucial to ensure that end users obtain accurate viscoelastic analysis results. Viscoelastic coagulation assays use reagents such as calcium chloride and various biomolecules, such as heparinase. These reagents can degrade over time, and results obtained using them can become inaccurate and unreliable.

[0032] Current quality control formulations used to test these reagents and devices containing them require human donors and contain human blood, human plasma, or components derived from human blood, such as human red blood cells. The use of quality control formulations containing blood components derived from human donors is relatively costly and poses an infection risk to the donor. Furthermore, the use of human-derived blood components is inconvenient for mass production.

[0033] The quality control formulations described herein, used to test the effectiveness of viscoelastic analysis reagents, do not require human donors for manufacture. In some embodiments, these quality control formulations contain citrated bovine plasma, recombinant human tissue factor, and heparin (preferably low molecular weight heparin). In some embodiments, these quality control formulations further include at least one additional component selected from the group consisting of buffers, stabilizers, and preservatives. Guidance on the use of embodiments of the quality control formulations described herein is provided in Appendix A, which is attached to this application and is incorporated herein by reference in its entirety.

[0034] The viscoelastic analysis methods and assays described herein utilize reagent-containing TEG Global Blood Coagulation Assay Cartridges suitable for use with TEG6s blood coagulation analyzers. However, as will be understood by those skilled in the art, the methods and formulations described herein may also be applied using different viscoelastic analysis methods and coagulation assays, and different viscoelastic analysis devices.

[0035] The TEG Global Blood Coagulation Assay Cartridge ("TEG Cartridge") is a citrate-enriched, multi-channel microfluidic cartridge that provides four coagulation assays demonstrating the functional measurement of coagulation factors, platelets, and fibrinogen, as well as fibrinosis. These four assays include: (i) the Kaolin Assay ("CK"), an intrinsic pathway activation assay that identifies hemostatic properties, bleeding risk, and thrombosis risk; (ii) the Kaolin and Heparinase Assay ("CKH"), which eliminates the influence of heparin in the test sample and, when used in conjunction with CK, assesses the presence of heparin or heparinoids in the system; and (iii) the RAPIDTEG, which incorporates both kaolin and tissue factor. TM (iv) The assay ("CRT"), which is activated via the intrinsic and extrinsic pathways, allows for a more rapid assessment of a patient's blood coagulation ability; and (iv) the functional fibrinogen assay ("CFF"), which is activated via the extrinsic pathway, uses GPIIb / IIIa platelet inhibitors to isolate the fibrin contribution to coagulation strength and, when used with CK, assesses the relative contribution of platelets and fibrinogen to overall coagulation strength.

[0036] In some embodiments, the quality control formulation of the present invention results in an extension of both the R time of CK and the R time of CKH, indicating that the amount / activity of CaCl2 in the TEG cartridge has decreased to such an extent that the cartridge, and other cartridges of the same lot, can no longer produce reliable results and are therefore unusable for testing patient samples.

[0037] In some embodiments, heparin is included in the quality control formulation to separate the R values ​​of CK and CKH. If the R value of CKH increases and becomes closer to the R value of CK, it indicates that the amount / activity of heparinase in the cartridge has decreased to such an extent that that cartridge, and other cartridges in the same lot, can no longer produce reliable results and are therefore unusable for testing patient samples.

[0038] In some embodiments, methods are provided for preparing a quality control formulation having target concentrations of citrated bovine plasma, human tissue factor, and heparin in order to test the effectiveness of at least one viscoelastic analytical reagent.

[0039] In some embodiments, the method includes determining a target concentration of citrated bovine plasma for a formulation by measuring a first viscoelastic property having a target value for a citrated bovine plasma dilution and obtaining a first value for the citrated bovine plasma dilution. Based on this first value, a predicted citrated bovine plasma concentration is determined corresponding to the first viscoelastic property target value. This predicted citrated bovine plasma concentration is the target citrated bovine plasma concentration.

[0040] This method further includes determining the target concentration of human tissue factor for a formulation by measuring a second viscoelastic property having a target value for a human tissue factor dilution and obtaining a second value for the human tissue factor dilution. Based on this second value, a predicted human tissue factor concentration is determined corresponding to the second viscoelastic property target value. This predicted human tissue factor concentration is the target human tissue factor concentration.

[0041] This method also includes determining the target concentration of heparin for the formulation by measuring a third viscoelastic property having a target value for the heparin dilution and obtaining a third value for the heparin dilution. Based on this third value, a predicted heparin concentration is determined corresponding to the target value of the third viscoelastic property. This predicted heparin concentration is the target heparin concentration.

[0042] The formulation is then prepared by combining citrated bovine plasma, human tissue factor, and heparin so that the formulation contains citrated bovine plasma at a target citrated bovine plasma concentration, human tissue factor at a target human tissue factor concentration, and heparin at a target heparin concentration.

[0043] In some embodiments, the method comprises determining a target concentration of citrated bovine plasma for a formulation by preparing a citrated bovine plasma dilution series, the citrated bovine plasma dilution series having at least two members. A first viscoelastic property having a target value is measured for each member of the citrated bovine plasma dilution series, and a first value is obtained for each member of the citrated bovine plasma dilution series. A linear regression is calculated from the first values ​​for each member of the citrated bovine plasma dilution series, and a predicted citrated bovine plasma concentration corresponding to the target value of the first viscoelastic property is extrapolated from this linear regression. This predicted citrated bovine plasma concentration is the target citrated bovine plasma concentration.

[0044] This method further includes determining the target concentration of human tissue factor for a formulation by preparing a human tissue factor dilution series, the human tissue factor dilution series having at least two members. A second viscoelastic property having a target value is measured for each member of the human tissue factor dilution series, and a second value is obtained for each member of the human tissue factor dilution series. A linear regression is calculated from the second values ​​for each member of the human tissue factor dilution series, and a predicted human tissue factor concentration corresponding to the target value of the second viscoelastic property is extrapolated from this linear regression. This predicted human tissue factor concentration is the target human tissue factor concentration.

[0045] This method also involves determining the target concentration of heparin for a formulation by preparing a heparin dilution series, the heparin dilution series having at least two components. A third viscoelastic property having a target value is measured for each component of the heparin dilution series, and a third value is obtained for each component of the heparin dilution series. A linear regression is calculated from the third value for each component of the heparin dilution series, and a predicted heparin concentration corresponding to the target value of the third viscoelastic property is extrapolated from this linear regression. This predicted heparin concentration is the target heparin concentration.

[0046] The formulation is then prepared by combining citrated bovine plasma, human tissue factor, and heparin so that the formulation contains citrated bovine plasma at a target citrated bovine plasma concentration, human tissue factor at a target human tissue factor concentration, and heparin at a target heparin concentration.

[0047] When determining the target concentration of citrated bovine plasma, the concentrations of citrated bovine plasma in each component of the citrated bovine plasma dilution series differ, but the concentration of human tissue factor in each component of the citrated bovine plasma dilution series remains constant.

[0048] When determining the target concentration of human tissue factor, the concentrations of human tissue factor in each member of the human tissue factor dilution series differ, but each member of the human tissue factor dilution series has a citrate-added bovine plasma concentration that is the target citrate-added bovine plasma concentration, and also has a constant concentration of heparin. In some embodiments, the constant concentration of heparin in each member of the human tissue factor dilution series is the target heparin concentration.

[0049] When determining the target concentration of heparin, although the heparin concentrations of each component in the heparin dilution series are different, each component in the heparin dilution series has a citrate-added bovine plasma concentration that is the target citrate-added bovine plasma concentration, and also has a constant concentration of human tissue factor. In some embodiments, the constant concentration of human tissue factor in each component of the heparin dilution series is the target human tissue factor concentration.

[0050] In some embodiments, the target concentrations of human tissue factor and heparin may be determined simultaneously by preparing a human tissue factor dilution series, each member of which has a different human tissue factor concentration and a citrate-bovine plasma concentration that is the target citrate-bovine plasma concentration. Each member of the human tissue factor dilution series may then be divided into several sub-members, each sub-member derived from a particular member having a different heparin concentration.

[0051] One or more of the first, second, and third viscoelastic properties may be measured using a viscoelastic analysis device. The viscoelastic analysis device may include a microfluidic cartridge.

[0052] In some embodiments, the first viscoelastic property is the MA value, and the target value for the first viscoelastic property is approximately 63. The first viscoelastic property and the first target value can be determined from a CK coagulation assay.

[0053] In some embodiments, at least one of the second and third viscoelastic properties is an R value, and at least one of the second and third viscoelastic properties has a target value of about 6. In some embodiments, at least one of the second and third viscoelastic properties has a target value of about 4. One or more of the second and third viscoelastic properties, and one or more of the second and third viscoelastic target values ​​can be determined from CK coagulation assays and CKH coagulation assays. When evaluating a heparin-containing formulation using a heparinase-intensive coagulation assay (such as CK), the target value for the viscoelastic property R value is about 6. When evaluating a heparin-containing formulation using a heparinase-intensive coagulation assay (such as CKH), the target value for the viscoelastic property R value is 4. When evaluating a heparin-deficient formulation, the target value for the viscoelastic property R value is about 4.

[0054] In some embodiments, a quality control formulation is provided for testing the effectiveness of at least one viscoelastic analysis reagent, the formulation comprising 60–90% citrated bovine plasma, 0.05–0.20% human tissue factor, and 0.05–0.30 U / ml heparin.

[0055] The quality control formulation may further contain stabilizers well known in the art, such as glycine, HEPES buffer, and other buffers well known in the art.

[0056] The quality control formulation may further contain one or more preservatives known in the art, such as sodium azide.

[0057] In some embodiments, the quality control formulation has a pH of about 7.3 to about 7.9. The total protein content of the formulation is preferably greater than about 6.4 g / dL. [Examples]

[0058] Example 1: Preparation of Quality Control Formula

[0059] A sample volume was taken from a lot of citrated bovine plasma (BCP) (3.8% sodium citrate) and thawed in a warm water bath. 5 ml of physiological saline (HBS) buffered with 1 M HEPES was added to 500 ml of BCP to obtain a final HEPES concentration of 9.9 mM. The resulting solution was mixed for 5 minutes.

[0060] Next, a series of test formulations were prepared by varying the concentration of BCP and keeping the recombinant human tissue factor, recombiplastin 2G (R2G), at a constant concentration (0.05%). Viscoelastic analysis was performed on each formulation using a TEG6s analyzer with a TEG Global blood coagulation cartridge, and the data is shown in Table 1 below.

[0061] [Table 1]

[0062] Based on linear regression of CK MA value data, the target BCP concentration is predicted to be 83.6%, corresponding to the target value (63) for the viscoelastic properties (MA value) of the CK assay.

[0063] Next, the final BCP concentration is adjusted to the target BCP concentration (~83.6% based on linear regression, as described above) using a dilution of R2G (using a 25% R2G working solution of R2G) and LOVENOX. 登録商標 (Heparin) Dilution (10U / ml LOVENOX) 登録商標 A series of dilutions (using the working solution) were prepared. These dilutions, containing other buffers and stabilizers, are shown in Table 2.

[0064] [Table 2]

[0065] R2G / LOVENOX from Table 2 登録商標1 mL of each of the dilutions was aliquoted into separate glass vials. The vials were frozen at ≤ -35 °C and the samples were lyophilized. When the samples were lyophilized, the lyophilized R2G / LOVENOX 登録商標 Each of the dilutions was reconstituted with distilled deionized water.

[0066] Reconstituted R2G / LOVENOX 登録商標 For each of the dilutions, viscoelastic analysis was performed on a TEG6s analyzer using a TEG global blood coagulation cartridge.

[0067] Reconstituted R2G / LOVENOX 登録商標 From the viscoelastic analysis of the dilutions, dilution number 10 corresponding to an R2G concentration of 0.075% and a LOVENOX 登録商標 concentration of 0.100 U / mL resulted in a CK R value closest to the nominal CK assay R target value of 6.0 shown in Table 3, and at the same time produced additional viscoelastic analysis results that fell within the acceptance range shown in Table 4.

[0068]

Table 3

[0069]

Table 4

[0070] Based on the results presented above, a quality control formulation with a target BCP concentration of ~83.6%, a target R2G concentration of 0.075%, and a target LOVENOX 登録商標 concentration of 0.100 U / mL was selected and the manufacturing process was advanced.

[0071] Example 2: Use of the Quality Control Formulation

[0072] The quality control formulation described herein can be used to detect degradation of cartridge viscoelastic analysis reagents. Figures 1 and 2 show a comparison of the responses of human donor blood samples and samples of the quality control formulation described herein, using TEG cartridges with different simulated CaCl2 (essential component) degradation levels. Clearly, the quality control formulation can detect CaCl2 degradation at 50%, and above that level, the R value time in the CKH channel is significantly prolonged, and no coagulation occurs in the CK channel.

[0073] The quality control formulation can also detect heparinase degradation in the TEG cartridge (see Figure 3), which is indicated by an increase in the R value of CKH to match the R value of CK when the simulated degradation level exceeds 90%.

[0074] Various embodiments of the present invention may be characterized by potential claims enumerated in paragraphs following this paragraph (and prior to the actual claims presented at the end of the application). These potential claims form part of the detailed description of the present invention. Accordingly, the features of the following potential claims may be presented as actual claims in subsequent proceedings relating to the present application or any application claiming priority thereunder. The inclusion of such potential claims should not be construed as meaning that the actual claims do not encompass the features of the potential claims. Accordingly, a decision not to present these potential claims in subsequent proceedings should not be construed as providing those features in general.

[0075] Without limitation, potential claimable features (preceded by the letter "P" to avoid confusion with the actual claims presented later) include: P1. A formulation for testing the efficacy of at least one viscoelastic analysis reagent, comprising a method for preparing a formulation having a target citrate-added bovine plasma concentration, a target human tissue factor concentration, and a target heparin concentration: A series of citrate-added bovine plasma dilutions is prepared, and this series of citrate-added bovine plasma dilutions has at least two members. For each component of the citrate-added bovine plasma dilution series, a first viscoelastic property is measured, and a first value is obtained for each component of the citrate-added bovine plasma dilution series. This first viscoelastic property has a target value. The first linear regression is calculated from the first value of each component in the citrate-added bovine plasma dilution series, and The target concentration of citrated bovine plasma for the formulation is determined by extrapolating the predicted citrated bovine plasma concentration corresponding to the first viscoelastic property target value from the first linear regression, and ensuring that this predicted citrated bovine plasma concentration is the target citrated bovine plasma concentration: A human tissue factor dilution series is prepared, and this human tissue factor dilution series has at least two members. The second viscoelastic property is measured for each component of the human tissue factor dilution series, and a second value is obtained for each component of the human tissue factor dilution series. This second viscoelastic property has a target value. We calculate the second linear regression from the second value of each component of the human tissue factor dilution series, and From this second linear regression, the predicted human tissue factor concentration corresponding to the second viscoelastic property target value is extrapolated, and the target concentration of human tissue factor for the formulation is determined by the fact that this predicted human tissue factor concentration is the target human tissue factor concentration: A heparin dilution series is prepared, and this heparin dilution series has at least two members. A third viscoelastic property is measured for each component of the heparin dilution series, and a third value is obtained for each component of the heparin dilution series. This third viscoelastic property has a target value. We calculate a third linear regression from the third value of each component of the heparin dilution series, and From this third linear regression, we extrapolate the predicted heparin concentration corresponding to the third viscoelastic property target value, and by ensuring that this predicted heparin concentration is the target heparin concentration, we determine the target concentration of heparin for the formulation: and A method comprising preparing a formulation by combining citrate-bovine plasma, human tissue factor, and heparin so that the formulation contains citrate-bovine plasma at a target citrate-bovine plasma concentration, human tissue factor at a target human tissue factor concentration, and heparin at a target heparin concentration. P2. A method for preparing a compound according to claim P1, In procedure (a), Each member of the citrate-bovine plasma dilution series has a citrate-bovine plasma concentration, and the concentration of citrate-bovine plasma in any one member of the citrate-bovine plasma dilution series is different from the concentration of citrate-bovine plasma in any other member of the citrate-bovine plasma dilution series, and Each component of the citrate-added bovine plasma dilution series has a human tissue factor concentration, and the human tissue factor concentrations of each component of the citrate-added bovine plasma dilution series are the same; In procedure (b), Each member of the human tissue factor dilution series has a human tissue factor concentration, and the concentration of human tissue factor in any one member of the human tissue factor dilution series is different from the concentration of human tissue factor in any other member of the human tissue factor dilution series. Each component of the human tissue factor dilution series has a citrated bovine plasma concentration, and the citrated bovine plasma concentration of each component of the human tissue factor dilution series is the target citrated bovine plasma concentration, and Each member of the human tissue factor dilution series has a heparin concentration, and the heparin concentrations of each member of the human tissue factor dilution series are the same; and In procedure (c), Each component of the heparin dilution series has a heparin concentration, and the heparin concentration in any one component of the heparin dilution series is different from the heparin concentration in any other component of the heparin dilution series. Each component of the heparin dilution series has a citrated bovine plasma concentration, and the citrated bovine plasma concentration of each component of the heparin dilution series is the target citrated bovine plasma concentration, and Each component of the heparin dilution series has a human tissue factor concentration, and the human tissue factor concentration of each component of the heparin dilution series is the target human tissue factor concentration. P3. A method for preparing a compound according to claim P1, In procedure (a), Each member of the citrate-bovine plasma dilution series has a citrate-bovine plasma concentration, and the concentration of citrate-bovine plasma in any one member of the citrate-bovine plasma dilution series is different from the concentration of citrate-bovine plasma in any other member of the citrate-bovine plasma dilution series, and Each component of the citrate-added bovine plasma dilution series has a human tissue factor concentration, and the human tissue factor concentrations of each component of the citrate-added bovine plasma dilution series are the same; In procedure (c), Each component of the heparin dilution series has a heparin concentration, and the heparin concentration in any one component of the heparin dilution series is different from the heparin concentration in any other component of the heparin dilution series. Each component of the heparin dilution series has a citrated bovine plasma concentration, and the citrated bovine plasma concentration of each component of the heparin dilution series is the target citrated bovine plasma concentration, and Each component of the heparin dilution series has a human tissue factor concentration, and the human tissue factor concentrations of each component of the heparin dilution series are the same; and In procedure (b), Each member of the human tissue factor dilution series has a human tissue factor concentration, and the concentration of human tissue factor in any one member of the human tissue factor dilution series is different from the concentration of human tissue factor in any other member of the human tissue factor dilution series. Each component of the human tissue factor dilution series has a citrated bovine plasma concentration, and the citrated bovine plasma concentration of each component of the human tissue factor dilution series is the target citrated bovine plasma concentration, and Each component of the human tissue factor dilution series has a heparin concentration, and the heparin concentration of each component of the human tissue factor dilution series is the target heparin concentration. P4. A formulation for testing the efficacy of at least one viscoelastic analysis reagent, comprising a method for preparing a formulation having target concentrations of citrated bovine plasma, human tissue factor, and heparin: The first viscoelastic properties of the citrate-added bovine plasma dilution were measured to obtain a first value for the citrate-added bovine plasma dilution, and this first viscoelastic property had the target value, and Based on the first value, the predicted citrate-added bovine plasma concentration is determined corresponding to the first viscoelasticity target value, and the predicted citrate-added bovine plasma concentration is equal to the target citrate-added bovine plasma concentration; The target concentration of citrated bovine plasma for the formulation is determined by: The second viscoelastic property of the human tissue factor dilution was measured to obtain a second value for the human tissue factor dilution, and this second viscoelastic property had the target value, and Based on the second value, the predicted human tissue factor concentration is determined corresponding to the second viscoelastic property target value, and the predicted human tissue factor concentration is the target human tissue factor concentration; The target concentration of human tissue factor for the formulation is determined by: The third viscoelastic property of the heparin diluent was measured to obtain a third value for the heparin diluent, and this third viscoelastic property had the target value, and Based on the third value, the predicted heparin concentration is determined corresponding to the third viscoelastic property target value, and the predicted heparin concentration is the target heparin concentration; The target concentration of heparin for the formulation is determined by: and A method comprising preparing a formulation by combining citrate-bovine plasma, human tissue factor, and heparin such that the formulation contains citrate-bovine plasma at a target citrate-bovine plasma concentration, human tissue factor at a target human tissue factor concentration, and heparin at a target heparin concentration. P5. A formulation for testing the efficacy of at least one viscoelastic analysis reagent, comprising a method for preparing a formulation having a target citrate-added bovine plasma concentration, a target human tissue factor concentration, and a target heparin concentration: A series of citrate-added bovine plasma dilutions is prepared, and the series of citrate-added bovine plasma dilutions has at least two members. For each component of the citrate-added bovine plasma dilution series, a first viscoelastic property is measured, and a first value is obtained for each component of the citrate-added bovine plasma dilution series. This first viscoelastic property has a target value. The first linear regression is calculated from the first value of each component in the citrate-added bovine plasma dilution series, and Extrapolate the predicted citrate-added bovine plasma concentration corresponding to the first viscoelastic property target value from the first linear regression, such that the predicted citrate-added bovine plasma concentration is equal to the target citrate-added bovine plasma concentration; The target citrate-added bovine plasma concentration for the formulation is determined by: A set of human tissue factor dilutions is prepared, and the set of human tissue factor dilutions has at least two heparin dilution series, each heparin dilution series having at least two members, where each member of each heparin dilution series has a human tissue factor concentration and a heparin concentration. The second viscoelastic property is measured for each component of each heparin dilution series, and a second value is obtained for each component of each heparin dilution series, and this second viscoelastic property has a target value. Identify a selected member from the human tissue factor dilution set that has a measured second viscoelastic property closer to the second viscoelastic property target value than the other members of the human tissue factor dilution set, and ensure that the human tissue factor concentration of the selected member is the target human tissue factor concentration, and that the heparin concentration of the selected member is the target heparin concentration; The target human tissue factor concentration and target heparin concentration for the formulation are determined by: and A method comprising preparing a formulation by combining citrate-bovine plasma, human tissue factor, and heparin such that the formulation contains citrate-bovine plasma at a target citrate-bovine plasma concentration, human tissue factor at a target human tissue factor concentration, and heparin at a target heparin concentration. P6. A method for preparing a compound according to claim P5, In procedure (a), Each member of the citrate-bovine plasma dilution series has a citrate-bovine plasma concentration, and the concentration of citrate-bovine plasma in any one member of the citrate-bovine plasma dilution series is different from the concentration of citrate-bovine plasma in any other member of the citrate-bovine plasma dilution series, and Each component of the citrate-added bovine plasma dilution series has a human tissue factor concentration, and the human tissue factor concentrations of each component of the citrate-added bovine plasma dilution series are the same; and In procedure (b), Each component of each series of heparin dilutions has a citrated bovine plasma concentration, and the citrated bovine plasma concentration of each component of each heparin dilution series is the target citrated bovine plasma concentration. The concentration of human tissue factor in any one heparin dilution series differs from the concentration of human tissue factor in any other heparin dilution series. The concentrations of human tissue factor in all members of any one of the heparin dilution series are the same, and The concentration of heparin in any one component of any heparin dilution series is different from the concentration of heparin in any other component of the same heparin dilution series. P7. The method of any one of the preceding claims, wherein the first viscoelastic property is measured using a viscoelastic analysis device. P8. The method of any one of the preceding claims, wherein the second viscoelastic property is measured using a viscoelastic analysis device. P9. The method of any one of the preceding claims, wherein a third viscoelastic property is measured using a viscoelastic analysis device. P10. The method according to any one of the prior claims, wherein the device for viscoelastic analysis is a microfluidic cartridge. P11. The method of any one of the preceding claims, wherein the first viscoelastic property is the MA value. P12. The method of claim P11, wherein the first viscoelastic property target value is approximately 63. P13. The method of any one of the preceding claims, wherein the second viscoelastic property is the R value. P14. The method of claim P13, wherein the target value of the second viscoelastic property is approximately 6. P15. The method of any one of the preceding claims, wherein the third viscoelastic property is the R value. P16. The method of claim P15, wherein the target value of the third viscoelastic property is approximately 6. P17. The method of either claim P1 or P4, wherein steps (b) and (c) are performed simultaneously. P18. A formulation for testing the effectiveness of at least one viscoelastic analysis reagent: 60-90% citrated bovine plasma, 0.05-0.20% human tissue factor, and 0.05-0.30 U / ml heparin A compound containing the following ingredients. P19. The formulation of claim P18, further comprising a stabilizer. P20. The formulation of claim P19, wherein the stabilizer is selected from the group consisting of glycine, HEPES buffer, and combinations thereof. P21. A formulation according to any one of claims P18 to P20, further comprising a preservative. P22. The formulation of claim P21, wherein the preservative is sodium azide. P23. A formulation according to any one of claims P18 to P22, wherein the formulation has a pH of approximately 7.3 to approximately 7.9. P24. A formulation according to any one of claims P18 to P23, wherein the formulation has a total protein content greater than approximately 6.4 g / dL.

[0076] The embodiments of the present invention described above are intended to be illustrative only; many variations and modifications will be apparent to those skilled in the art. All such variations and modifications are intended to fall within the scope of the present invention as defined in the appended claims.

Claims

1. A formulation for testing the efficacy of at least one viscoelastic analysis reagent, comprising a method for preparing a formulation having a target citrate-added bovine plasma concentration, a target human tissue factor concentration, and a target heparin concentration: A series of citrate-added bovine plasma dilutions is prepared, and this series of citrate-added bovine plasma dilutions has at least two members. For each component of the citrate-added bovine plasma dilution series, a first viscoelastic property is measured, and a first value is obtained for each component of the citrate-added bovine plasma dilution series. This first viscoelastic property has a target value. The first linear regression is calculated from the first value of each component in the citrate-added bovine plasma dilution series, and Extrapolate the predicted citrate-added bovine plasma concentration corresponding to the first viscoelastic property target value from the first linear regression, and ensure that this predicted citrate-added bovine plasma concentration is the target citrate-added bovine plasma concentration; The target concentration of citrated bovine plasma for the formulation is determined by: A human tissue factor dilution series is prepared, and this human tissue factor dilution series has at least two members. The second viscoelastic property is measured for each component of the human tissue factor dilution series, and a second value is obtained for each component of the human tissue factor dilution series. This second viscoelastic property has a target value. The second linear regression was calculated from the second value of each component of the human tissue factor dilution series, and From this second linear regression, extrapolate the predicted human tissue factor concentration corresponding to the second viscoelastic property target value, and ensure that this predicted human tissue factor concentration is equal to the target human tissue factor concentration; The target concentration of human tissue factor for the formulation is determined by: A heparin dilution series is prepared, and this heparin dilution series has at least two members. The third viscoelastic property is measured for each component of the heparin dilution series, and a third value is obtained for each component of the heparin dilution series, and this third viscoelastic property has a target value. The third linear regression is calculated from the third value of each component of the heparin dilution series, and From this third linear regression, the predicted heparin concentration corresponding to the third viscoelastic property target value is extrapolated, and this predicted heparin concentration is the target heparin concentration; The target concentration of heparin for the formulation is determined by: and A method comprising preparing a formulation by combining citrate-bovine plasma, human tissue factor, and heparin such that the formulation contains citrate-bovine plasma at a target citrate-bovine plasma concentration, human tissue factor at a target human tissue factor concentration, and heparin at a target heparin concentration.

2. A method for preparing a compound according to claim 1, In procedure (a), Each component of the citrate-added bovine plasma dilution series has a citrate-added bovine plasma concentration, and the concentration of citrate-added bovine plasma in any one component of the citrate-added bovine plasma dilution series is different from the concentration of citrate-added bovine plasma in any other component of the citrate-added bovine plasma dilution series, and Each component of the citrate-added bovine plasma dilution series has a human tissue factor concentration, and the human tissue factor concentrations of each component of the citrate-added bovine plasma dilution series are the same; In procedure (b), Each member of the human tissue factor dilution series has a human tissue factor concentration, and the concentration of human tissue factor in any one member of the human tissue factor dilution series is different from the concentration of human tissue factor in any other member of the human tissue factor dilution series. Each component of the human tissue factor dilution series has a citrated bovine plasma concentration, and the citrated bovine plasma concentration of each component of the human tissue factor dilution series is the target citrated bovine plasma concentration, and Each member of the human tissue factor dilution series has a heparin concentration, and the heparin concentrations of each member of the human tissue factor dilution series are the same; and In procedure (c), Each component of the heparin dilution series has a heparin concentration, and the heparin concentration in any one component of the heparin dilution series is different from the heparin concentration in any other component of the heparin dilution series. Each component of the heparin dilution series has a citrated bovine plasma concentration, and the citrated bovine plasma concentration of each component of the heparin dilution series is the target citrated bovine plasma concentration, and Each component of the heparin dilution series has a human tissue factor concentration, and the human tissue factor concentration of each component of the heparin dilution series is the target human tissue factor concentration.

3. A method for preparing a compound according to claim 1, In procedure (a), Each component of the citrate-added bovine plasma dilution series has a citrate-added bovine plasma concentration, and the concentration of citrate-added bovine plasma in any one component of the citrate-added bovine plasma dilution series is different from the concentration of citrate-added bovine plasma in any other component of the citrate-added bovine plasma dilution series, and Each component of the citrate-added bovine plasma dilution series has a human tissue factor concentration, and the human tissue factor concentrations of each component of the citrate-added bovine plasma dilution series are the same; In procedure (c), Each component of the heparin dilution series has a heparin concentration, and the heparin concentration in any one component of the heparin dilution series is different from the heparin concentration in any other component of the heparin dilution series. Each component of the heparin dilution series has a citrated bovine plasma concentration, and the citrated bovine plasma concentration of each component of the heparin dilution series is the target citrated bovine plasma concentration, and Each component of the heparin dilution series has a human tissue factor concentration, and the human tissue factor concentrations of each component of the heparin dilution series are the same; and In procedure (b), Each member of the human tissue factor dilution series has a human tissue factor concentration, and the concentration of human tissue factor in any one member of the human tissue factor dilution series is different from the concentration of human tissue factor in any other member of the human tissue factor dilution series. Each component of the human tissue factor dilution series has a citrated bovine plasma concentration, and the citrated bovine plasma concentration of each component of the human tissue factor dilution series is the target citrated bovine plasma concentration, and Each component of the human tissue factor dilution series has a heparin concentration, and the heparin concentration of each component of the human tissue factor dilution series is the target heparin concentration.

4. A formulation for testing the effectiveness of at least one viscoelastic analysis reagent, comprising a method for preparing a formulation having target concentrations of citrated bovine plasma, human tissue factor, and heparin: The first viscoelastic properties of the citrate-added bovine plasma dilution are measured to obtain a first value for the citrate-added bovine plasma dilution, and this first viscoelastic property has the target value, and Based on the first value, the predicted citrate-added bovine plasma concentration is determined corresponding to the first viscoelasticity target value, and the predicted citrate-added bovine plasma concentration is equal to the target citrate-added bovine plasma concentration; The target concentration of citrated bovine plasma for the formulation is determined by: The second viscoelastic property of the human tissue factor dilution was measured to obtain a second value for the human tissue factor dilution, and this second viscoelastic property had the target value, and Based on the second value, the predicted human tissue factor concentration is determined in accordance with the second viscoelastic property target value, and the predicted human tissue factor concentration is the target human tissue factor concentration; The target concentration of human tissue factor for the formulation is determined by: The third viscoelastic property of the heparin diluent was measured to obtain a third value for the heparin diluent, and this third viscoelastic property had the target value, and Based on the third value, the predicted heparin concentration is determined corresponding to the third viscoelastic property target value, and the predicted heparin concentration is the target heparin concentration; The target concentration of heparin for the formulation is determined by: and A method comprising preparing a formulation by combining citrate-bovine plasma, human tissue factor, and heparin such that the formulation contains citrate-bovine plasma at a target citrate-bovine plasma concentration, human tissue factor at a target human tissue factor concentration, and heparin at a target heparin concentration.

5. A formulation for testing the efficacy of at least one viscoelastic analysis reagent, comprising a method for preparing a formulation having a target citrate-added bovine plasma concentration, a target human tissue factor concentration, and a target heparin concentration: A series of citrate-added bovine plasma dilutions is prepared, and the series of citrate-added bovine plasma dilutions has at least two members. For each component of the citrate-added bovine plasma dilution series, a first viscoelastic property is measured, and a first value is obtained for each component of the citrate-added bovine plasma dilution series. This first viscoelastic property has a target value. The first linear regression is calculated from the first value of each component in the citrate-added bovine plasma dilution series, and Extrapolate the predicted citrate-added bovine plasma concentration corresponding to the first viscoelastic property target value from the first linear regression, such that the predicted citrate-added bovine plasma concentration is equal to the target citrate-added bovine plasma concentration; The target citrate-added bovine plasma concentration for the formulation is determined by: A set of human tissue factor dilutions is prepared, and the set of human tissue factor dilutions has at least two heparin dilution series, each heparin dilution series having at least two members, where each member of each heparin dilution series has a human tissue factor concentration and a heparin concentration. The second viscoelastic property is measured for each component of each heparin dilution series, and a second value is obtained for each component of each heparin dilution series, and this second viscoelastic property has a target value. Identify a selected member from the human tissue factor dilution set that has a measured second viscoelastic property closer to the second viscoelastic property target value than the other members of the human tissue factor dilution set, and ensure that the human tissue factor concentration of the selected member is the target human tissue factor concentration, and that the heparin concentration of the selected member is the target heparin concentration; The target human tissue factor concentration and target heparin concentration for the formulation are determined by: and A method comprising preparing a formulation by combining citrate-bovine plasma, human tissue factor, and heparin such that the formulation contains citrate-bovine plasma at a target citrate-bovine plasma concentration, human tissue factor at a target human tissue factor concentration, and heparin at a target heparin concentration.

6. A method for preparing a compound according to claim 5, In procedure (a), Each component of the citrate-added bovine plasma dilution series has a citrate-added bovine plasma concentration, and the concentration of citrate-added bovine plasma in any one component of the citrate-added bovine plasma dilution series is different from the concentration of citrate-added bovine plasma in any other component of the citrate-added bovine plasma dilution series, and Each component of the citrate-added bovine plasma dilution series has a human tissue factor concentration, and the human tissue factor concentrations of each component of the citrate-added bovine plasma dilution series are the same; and In procedure (b), Each component of each series of heparin dilutions has a citrated bovine plasma concentration, and the citrated bovine plasma concentration of each component of each heparin dilution series is the target citrated bovine plasma concentration. The concentration of human tissue factor in any one heparin dilution series differs from the concentration of human tissue factor in any of the other heparin dilution series. The concentrations of human tissue factor in all members of any one of the heparin dilution series are the same, and The concentration of heparin in any one component of any heparin dilution series is different from the concentration of heparin in any other component of the same heparin dilution series.

7. The method according to any one of the preceding claims, wherein the first viscoelastic property is measured using a viscoelastic analysis device.

8. The method of any one of the preceding claims, wherein a second viscoelastic property is measured using a viscoelastic analysis device.

9. The method of any one of the preceding claims, wherein a third viscoelastic property is measured using a viscoelastic analysis device.

10. The method according to any one of the prior claims, wherein the device for viscoelastic analysis is a microfluidic cartridge.

11. The method according to any one of the preceding claims, wherein the first viscoelastic property is the MA value.

12. The method of claim 11, wherein the first viscoelastic property target value is approximately 63.

13. The method according to any one of the preceding claims, wherein the second viscoelastic property is the R value.

14. The method of claim 13, wherein the target value of the second viscoelastic property is approximately 6.

15. The method according to any one of the preceding claims, wherein the third viscoelastic property is the R value.

16. The method of claim 15, wherein the target value of the third viscoelastic property is approximately 6.

17. The method according to any one of claims 1 and 4, wherein steps (b) and (c) are performed simultaneously.

18. A formulation for testing the effectiveness of at least one viscoelastic analysis reagent: A composition comprising citrated bovine plasma, human tissue factor, and heparin, present in concentrations determined by any one of claims 1 to 17, to provide a target value for viscoelastic properties used in the method.

19. The formulation according to claim 18, further comprising a stabilizer.

20. The formulation of claim 19, wherein the stabilizer is selected from the group consisting of glycine, HEPES buffer, and combinations thereof.

21. Furthermore, a formulation according to any one of claims 18 to 20, which also contains a preservative.

22. The formulation according to claim 21, wherein the preservative is sodium azide.

23. The formulation according to any one of claims 18 to 22, wherein the formulation has a pH of approximately 7.3 to approximately 7.

9.

24. The formulation according to any one of claims 18 to 23, wherein the formulation has a total protein content greater than approximately 6.4 g / dL.

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