Reduces erythrocyte sedimentation rate

Abstract

The present invention relates to reducing the erythrocyte sedimentation rate in a blood sample. In particular, formulations, compositions, preparations, kits and methods for reducing the erythrocyte sedimentation rate in a blood sample are provided.

Description

[0001] This application is a divisional application based on a patent application with an application date of December 6, 2016, a priority date of December 8, 2015, an application number of 201680081392.6, and an invention name of "Reducing the Erythrocyte Sedimentation Rate".

[0002] Cross-references

[0003] This application claims the benefit of U.S. Provisional Application No. 62 / 264,786, filed December 8, 2015, which is incorporated herein by reference in its entirety. Background of the Invention

[0004] There is a need for improved formulations and methods for reducing the rate at which blood sediments for a period of time sufficient for storage, transportation, and shipment for research, diagnostic, and therapeutic purposes. The present invention generally relates to reducing the sedimentation rate of one or more red blood cells in a blood sample. In particular, the present invention relates to formulations, compositions, articles, kits, and methods for reducing the sedimentation rate of red blood cells in a blood sample. Summary of the invention

[0005] In some embodiments, described herein is an in vitro method for reducing the erythrocyte sedimentation rate in a blood sample, the method comprising: combining a blood sample with a certain amount of a preparation comprising sucralose, wherein the amount is sufficient to produce a treated blood sample having a sucralose concentration of at least about 5mM sucralose, thereby reducing the erythrocyte sedimentation rate compared to the erythrocyte sedimentation rate in an untreated blood sample. In some embodiments, the treated blood sample has a sucralose concentration of about 5mM sucralose to about 100mM sucralose. In some embodiments, the treated blood sample has a sucralose concentration of about 10mM sucralose to about 100mM sucralose. In some embodiments, the treated blood sample has a sucralose concentration of about 20mM sucralose to about 100mM sucralose. In some embodiments, the treated blood sample has a sucralose concentration of about 24mM sucralose to about 100mM sucralose. In some embodiments, the treated blood sample has a sucralose concentration of about 15mM sucralose to about 50mM sucralose. In some embodiments, the treated blood sample has a sucralose concentration of about 5mM sucralose to about 50mM sucralose. In some embodiments, the treated blood sample has a sucralose concentration of about 5mM sucralose to about 40mM sucralose. In some embodiments, the treated blood sample has a sucralose concentration of about 10mM sucralose to about 35mM sucralose. In some embodiments, the treated blood sample has a sucralose concentration of about 10mM sucralose to about 30mM sucralose. In some embodiments, the treated blood sample has a sucralose concentration of about 5mM sucralose to about 25mM sucralose. In some embodiments, the treated blood sample has a sucralose concentration of about 5mM sucralose up to but not including 25mM sucralose. In some embodiments, the treated blood sample has a sucralose concentration of about 5mM sucralose to about 20mM sucralose. In some embodiments, the treated blood sample has a sucralose concentration of about 5mM sucralose to about 15mM sucralose. In some embodiments, the treated blood sample has a sucralose concentration of about 5mM sucralose to about 10mM sucralose. In some embodiments, the treated blood sample has a sucralose concentration of about 24mM. In some embodiments, the treated blood sample has a sucralose concentration of about 25mM. In some embodiments, the erythrocyte sedimentation rate is reduced by at least 10%, at least 20%, at least 30%, at least 40%, at least 50%, at least 60%, at least 70%, at least 80%, at least 90%, at least 95% of the erythrocyte sedimentation rate of the untreated blood sample. In some embodiments, the formulation is in the form of a powder, solid, lyophilized form, solution or aqueous solution.In some embodiments, the preparation is a powder. In some embodiments, the preparation is a solid. In some embodiments, the preparation is lyophilized. In some embodiments, the preparation is a solution. In some embodiments, the solution is an aqueous solution. In some embodiments, the preparation consists of sucralose. In some embodiments, the preparation further comprises an anticoagulant. In some embodiments, the anticoagulant is selected from acid citrate dextrose solution (ACD), sodium heparin, sodium fluoride, lithium heparin, tripotassium ethylenediaminetetraacetate (K. 3 EDTA), ethylenediaminetetraacetic acid dipotassium (K 2 In some embodiments, the anticoagulant is an acid citrate dextrose solution (ACD). In some embodiments, the anticoagulant is sodium heparin. In some embodiments, the anticoagulant is sodium fluoride. In some embodiments, the anticoagulant is lithium heparin. In some embodiments, the anticoagulant is tripotassium ethylenediaminetetraacetate (K 3 In some embodiments, the anticoagulant is dipotassium ethylenediaminetetraacetate (K 2 In some embodiments, the anticoagulant is hirudin. In some embodiments, the anticoagulant is sodium polyanethol sulfonate (SPS). In some embodiments, the preparation is contained in a blood collection tube, and the combining step occurs in the blood collection tube. In some embodiments, the blood collection tube is an evacuated blood collection tube. In some embodiments, the blood is collected from a subject. In some embodiments, the subject is a mammal. In some embodiments, the subject is a human.

[0006] In some embodiments, the present invention describes an in vitro method for keeping one or more red blood cells suspended in a blood sample, the method comprising: combining a blood sample with a certain amount of a preparation comprising sucralose, wherein the amount is sufficient to produce a treated blood sample having a sucralose concentration of at least about 5mM sucralose, whereby the one or more red blood cells remain suspended for a period of at least 30 minutes compared to an untreated blood sample. In some embodiments, the one or more red blood cells remain suspended for at least 10 minutes, at least 20 minutes, at least 30 minutes, at least 40 minutes, at least 50 minutes, at least 60 minutes, at least 90 minutes, at least 2 hours, at least 3 hours, at least 4 hours, at least 5 hours, at least 6 hours, at least 7 hours, at least 8 hours, at least 10 hours, at least 12 hours, at least 24 hours, or at least 48 hours. In some embodiments, the treated blood sample has a sucralose concentration of about 5mM sucralose to about 100mM sucralose. In some embodiments, the treated blood sample has a sucralose concentration of about 10mM sucralose to about 100mM sucralose. In some embodiments, the treated blood sample has a sucralose concentration of about 20mM sucralose to about 100mM sucralose. In some embodiments, the treated blood sample has a sucralose concentration of about 24mM sucralose to about 100mM sucralose. In some embodiments, the treated blood sample has a sucralose concentration of about 15mM sucralose to about 50mM sucralose. In some embodiments, the treated blood sample has a sucralose concentration of about 5mM sucralose to about 50mM sucralose. In some embodiments, the treated blood sample has a sucralose concentration of about 5mM sucralose to about 40mM sucralose. In some embodiments, the treated blood sample has a sucralose concentration of about 10mM sucralose to about 35mM sucralose. In some embodiments, the treated blood sample has a sucralose concentration of about 10mM sucralose to about 30mM sucralose. In some embodiments, the treated blood sample has a sucralose concentration of about 5mM sucralose to about 25mM sucralose. In some embodiments, the treated blood sample has a sucralose concentration of about 5mM sucralose up to but not including 25mM sucralose. In some embodiments, the treated blood sample has a sucralose concentration of about 5mM sucralose to about 20mM sucralose. In some embodiments, the treated blood sample has a sucralose concentration of about 5mM sucralose to about 15mM sucralose. In some embodiments, the treated blood sample has a sucralose concentration of about 5mM sucralose to about 10mM sucralose. In some embodiments, the treated blood sample has a sucralose concentration of about 24mM.In some embodiments, the treated blood sample has a sucralose concentration of about 25mM. In some embodiments, compared with untreated blood samples, at least 10%, at least 20%, at least 30%, at least 40%, at least 50%, at least 60%, at least 70%, at least 80%, at least 90%, at least 95% of the one or more red blood cells in the treated blood sample remain suspended. In some embodiments, the preparation is a powder. In some embodiments, the preparation is a solid. In some embodiments, the preparation is lyophilized. In some embodiments, the preparation is a solution. In some embodiments, the solution is an aqueous solution. In some embodiments, the preparation consists of sucralose. In some embodiments, the preparation further comprises an anticoagulant. In some embodiments, the anticoagulant is selected from acid citrate dextrose solution (ACD), sodium heparin, sodium fluoride, lithium heparin, tripotassium ethylenediaminetetraacetic acid (K. 3 EDTA), ethylenediaminetetraacetic acid dipotassium (K 2 In some embodiments, the anticoagulant is an acid citrate dextrose solution (ACD). In some embodiments, the anticoagulant is sodium heparin. In some embodiments, the anticoagulant is sodium fluoride. In some embodiments, the anticoagulant is lithium heparin. In some embodiments, the anticoagulant is tripotassium ethylenediaminetetraacetate (K 3 In some embodiments, the anticoagulant is dipotassium ethylenediaminetetraacetate (K 2 In some embodiments, the anticoagulant is hirudin. In some embodiments, the anticoagulant is sodium polyanethol sulfonate (SPS). In some embodiments, the preparation is contained in a blood collection tube, and the combining step occurs in the blood collection tube. In some embodiments, the blood collection tube is an evacuated blood collection tube. In some embodiments, the blood sample is collected from a subject. In some embodiments, the subject is a mammal. In some embodiments, the subject is a human.

[0007] In some embodiments, compositions comprising a blood sample and sucralose are described herein, wherein the concentration of the sucralose is from about 5mM sucralose to about 100mM sucralose. In some embodiments, the concentration of the sucralose is from about 10mM sucralose to about 100mM sucralose. In some embodiments, the concentration of the sucralose is from about 20mM sucralose to about 100mM sucralose. In some embodiments, the concentration of the sucralose is from about 24mM sucralose to about 100mM sucralose. In some embodiments, the concentration of the sucralose is from about 15mM sucralose to about 50mM sucralose. In some embodiments, the concentration of the sucralose is from about 5mM sucralose to about 50mM sucralose. In some embodiments, the concentration of the sucralose is from about 5mM sucralose to about 40mM sucralose. In some embodiments, the treated blood sample has a sucralose concentration of about 10mM sucralose to about 35mM sucralose. In some embodiments, the treated blood sample has a sucralose concentration of about 10mM sucralose to about 30mM sucralose. In some embodiments, the concentration of the sucralose is about 5mM sucralose up to but not including 25mM sucralose. In some embodiments, the concentration of the sucralose is about 5mM sucralose to about 20mM sucralose. In some embodiments, the concentration of the sucralose is about 5mM sucralose to about 15mM sucralose. In some embodiments, the concentration of the sucralose is about 5mM sucralose to about 10mM sucralose. In some embodiments, the treated blood sample has a sucralose concentration of about 24mM. In some embodiments, the treated blood sample has a sucralose concentration of about 25mM. In some embodiments, the composition further comprises an anticoagulant. In some embodiments, the anticoagulant is selected from acid citrate dextrose solution (ACD), sodium heparin, sodium fluoride, lithium heparin, tripotassium ethylenediaminetetraacetate (K 3 EDTA), ethylenediaminetetraacetic acid dipotassium (K 2 In some embodiments, the anticoagulant is an acid citrate dextrose solution (ACD). In some embodiments, the anticoagulant is sodium heparin. In some embodiments, the anticoagulant is sodium fluoride. In some embodiments, the anticoagulant is lithium heparin. In some embodiments, the anticoagulant is tripotassium ethylenediaminetetraacetate (K 3 In some embodiments, the anticoagulant is dipotassium ethylenediaminetetraacetate (K 2In some embodiments, the anticoagulant is hirudin. In some embodiments, the anticoagulant is sodium polyanethol sulfonate (SPS). In some embodiments, the composition is contained in a blood collection tube. In some embodiments, the blood collection tube is an evacuated blood collection tube. In some embodiments, the blood is collected from a subject. In some embodiments, the subject is a mammal. In some embodiments, the subject is a human.

[0008] In some embodiments, described herein are products comprising sucralose contained in a blood collection tube, wherein the amount of sucralose is sufficient to produce a final concentration of about 5 mM sucralose to about 100 mM sucralose in a blood sample. In some embodiments, the amount of sucralose is sufficient to produce a final concentration of about 10 mM sucralose to about 100 mM sucralose. In some embodiments, the amount of sucralose is sufficient to produce a final concentration of about 20 mM sucralose to about 100 mM sucralose. In some embodiments, the amount of sucralose is sufficient to produce a final concentration of about 24 mM sucralose to about 100 mM sucralose. In some embodiments, the amount of sucralose is sufficient to produce a final concentration of about 15 mM sucralose to about 50 mM sucralose. In some embodiments, the amount of sucralose is sufficient to produce a final concentration of about 5 mM sucralose to about 50 mM sucralose in a blood sample. In some embodiments, the amount of sucralose is sufficient to produce a final concentration of about 5mM sucralose to about 40mM sucralose in the blood sample. In some embodiments, the treated blood sample has a sucralose concentration of about 10mM sucralose to about 35mM sucralose. In some embodiments, the treated blood sample has a sucralose concentration of about 10mM sucralose to about 30mM sucralose. In some embodiments, the amount of sucralose is sufficient to produce a final concentration of about 5mM sucralose up to but not including 25mM sucralose in the blood sample. In some embodiments, the amount of sucralose is sufficient to produce a final concentration of about 5mM sucralose to about 20mM sucralose in the blood sample. In some embodiments, the amount of sucralose is sufficient to produce a final concentration of about 5mM sucralose to about 15mM sucralose in the blood sample. In some embodiments, the amount of sucralose is sufficient to produce a final concentration of about 5mM sucralose to about 10mM sucralose in the blood sample. In some embodiments, the treated blood sample has a sucralose concentration of about 24mM. In some embodiments, the treated blood sample has a sucralose concentration of about 25mM. In some embodiments, the sucralose is a powder. In some embodiments, the sucralose is a solid. In some embodiments, the sucralose is lyophilized. In some embodiments, the sucralose is in solution. In some embodiments, the sucralose solution is an aqueous solution. In some embodiments, the blood collection tube is an evacuated blood collection tube. In some embodiments, the product further comprises an anticoagulant. In some embodiments, the anticoagulant is selected from acid citrate dextrose solution (ACD), sodium heparin, sodium fluoride, lithium heparin, tripotassium ethylenediaminetetraacetate (K 3 EDTA), ethylenediaminetetraacetic acid dipotassium (K 2In some embodiments, the anticoagulant is an acid citrate dextrose solution (ACD). In some embodiments, the anticoagulant is sodium heparin. In some embodiments, the anticoagulant is sodium fluoride. In some embodiments, the anticoagulant is lithium heparin. In some embodiments, the anticoagulant is tripotassium ethylenediaminetetraacetate (K 3 In some embodiments, the anticoagulant is dipotassium ethylenediaminetetraacetate (K 2 In some embodiments, the anticoagulant is hirudin. In some embodiments, the anticoagulant is sodium polyanethol sulfonate (SPS).

[0009] In some embodiments, described herein are kits comprising an article of manufacture provided herein and a package insert.

[0010] This application includes the following implementation schemes:

[0011] Embodiment 1. An in vitro method for reducing the erythrocyte sedimentation rate in a blood sample, comprising: combining the blood sample with an amount of a formulation comprising sucralose, the amount being sufficient to produce a treated blood sample having a sucralose concentration of about 5 mM to about 50 mM sucralose, thereby reducing the erythrocyte sedimentation rate compared to the erythrocyte sedimentation rate in an untreated blood sample.

[0012] Embodiment 2. The method of embodiment 1, wherein the treated blood sample has a sucralose concentration of about 5 mM up to 25 mM sucralose.

[0013] Embodiment 3. A method according to any one of embodiments 1-2, wherein the erythrocyte sedimentation rate is reduced by at least 10%, at least 20%, at least 30%, at least 40%, at least 50%, at least 60%, at least 70%, at least 80%, at least 90%, at least 95% of the erythrocyte sedimentation rate of the untreated blood sample.

[0014] Embodiment 4. The method according to any one of embodiments 1-3, wherein the formulation is in the form of a powder, a solid, a lyophilized form, a solution or an aqueous solution.

[0015] Embodiment 5. The method of any one of embodiments 1-4, wherein the formulation consists of sucralose.

[0016] Embodiment 6. The method according to any one of embodiments 1-4, wherein the formulation further comprises an anticoagulant.

[0017] Embodiment 7. The method according to embodiment 6, wherein the anticoagulant is selected from the group consisting of acid citrate dextrose solution (ACD), sodium heparin, sodium fluoride, lithium heparin, tripotassium ethylenediaminetetraacetate (K 3 EDTA), ethylenediaminetetraacetic acid dipotassium (K 2 EDTA), hirudin and sodium polyanethol sulfonate (SPS).

[0018] Embodiment 8. The method of any one of Embodiments 1-7, wherein the formulation is contained within a blood collection tube and the combining step occurs within the blood collection tube.

[0019] Embodiment 9. The method according to embodiment 8, wherein the blood collection tube is an evacuated blood collection tube.

[0020] Embodiment 10. An in vitro method for maintaining one or more red blood cells in suspension in a blood sample, comprising: combining the blood sample with an amount of a formulation comprising sucralose, the amount sufficient to produce a treated blood sample having a sucralose concentration of about 5 mM to about 50 mM, whereby the one or more red blood cells remain suspended for a period of at least 30 minutes compared to an untreated blood sample.

[0021] Embodiment 11. The method of embodiment 10, wherein the treated blood sample has a sucralose concentration of about 5 mM up to 25 mM sucralose.

[0022] Embodiment 12. A method according to any of embodiments 10-11, wherein the one or more red blood cells remain suspended for a period of at least 10 minutes, at least 20 minutes, at least 30 minutes, at least 40 minutes, at least 50 minutes, at least 1 hour, at least 2 hours, at least 3 hours, at least 4 hours, at least 5 hours, at least 6 hours, at least 7 hours, at least 8 hours, at least 10 hours, at least 12 hours, at least 24 hours, or at least 48 hours.

[0023] Embodiment 13. The method of any one of embodiments 10-12, wherein the treated blood sample has a sucralose concentration of about 5 mM sucralose to about 50 mM sucralose.

[0024] Embodiment 14. A method according to any of embodiments 10-13, wherein at least 10%, at least 20%, at least 30%, at least 40%, at least 50%, at least 60%, at least 70%, at least 80%, at least 90%, at least 95% of the one or more red blood cells in the treated blood sample remain suspended compared to the untreated blood sample.

[0025] Embodiment 15. The method according to any one of embodiments 10-14, wherein the formulation is in the form of a powder, a solid, a lyophilized form, a solution or an aqueous solution.

[0026] Embodiment 16. The method of any one of embodiments 10-15, wherein the formulation consists of sucralose.

[0027] Embodiment 17. The method of any one of embodiments 10-15, wherein the formulation further comprises an anticoagulant.

[0028] Embodiment 18. The method according to embodiment 17, wherein the anticoagulant is selected from the group consisting of acid citrate dextrose solution (ACD), sodium heparin, sodium fluoride, lithium heparin, tripotassium ethylenediaminetetraacetate (K 3 EDTA), ethylenediaminetetraacetic acid dipotassium (K 2 EDTA), hirudin and sodium polyanethol sulfonate (SPS).

[0029] Embodiment 19. The method of any one of embodiments 10-18, wherein the formulation is contained within a blood collection tube and the combining step occurs within the blood collection tube.

[0030] Embodiment 20. The method according to embodiment 19, wherein the blood collection tube is an evacuated blood collection tube.

[0031] Embodiment 21. A composition comprising a blood sample and sucralose, wherein the concentration of sucralose is from about 5 mM sucralose up to but not including 50 mM sucralose.

[0032] Embodiment 22. The composition of embodiment 21, wherein the concentration of sucralose is from about 5 mM sucralose up to but not including 25 mM sucralose.

[0033] Embodiment 23. The composition according to embodiment 21, further comprising an anticoagulant.

[0034] Embodiment 24. The composition according to embodiment 23, wherein the anticoagulant is selected from acid citrate dextrose solution (ACD), sodium heparin, sodium fluoride, lithium heparin, tripotassium ethylenediaminetetraacetate (K 3 EDTA), ethylenediaminetetraacetic acid dipotassium (K 2 EDTA), hirudin and sodium polyanethol sulfonate (SPS).

[0035] Embodiment 25. The composition of embodiment 21, wherein the blood is human blood.

[0036] Embodiment 26. An article of manufacture comprising a blood collection tube comprising sucralose in an amount sufficient to produce a final concentration of about 5 mM sucralose to about 50 mM sucralose in a blood sample.

[0037] Embodiment 27. The article of manufacture of embodiment 26, wherein the blood collection tube is an evacuated blood collection tube.

[0038] Embodiment 28. The article of any one of Embodiments 26-27, wherein the blood collection tube further comprises an anticoagulant.

[0039] Embodiment 29. The article of manufacture according to embodiment 28, wherein the anticoagulant is selected from the group consisting of acid citrate dextrose solution (ACD), sodium heparin, sodium fluoride, lithium heparin, tripotassium ethylenediaminetetraacetate (K 3 EDTA), ethylenediaminetetraacetic acid dipotassium (K 2 EDTA), hirudin and sodium polyanethol sulfonate (SPS)

[0040] Incorporation by reference

[0041] All publications, patents, and patent applications mentioned in this specification are herein incorporated by reference to the same extent as if each individual publication, patent, or patent application was specifically and individually indicated to be incorporated by reference. BRIEF DESCRIPTION OF THE DRAWINGS

[0042] The novel features of the present invention are particularly set forth in the appended claims. A better understanding of the features and advantages of the present invention will be obtained by referring to the following detailed description of illustrative embodiments utilizing the principles of the present invention and the accompanying drawings, in which:

[0043] Figures 1A-1C Graph showing the reduction in erythrocyte sedimentation rate in whole blood after addition of 0.5 M sucralose, PBS or saline. Storage duration 0 hours ( Figure 1A ), 6 hours( Figure 1B ) and 24 hours ( Figure 1C ). NF = no preparation control.

[0044] Figure 2 The graph shows the sedimentation rate of red blood cells in whole blood after the addition of the indicated sucralose solutions, the indicated sugars, PBS or saline. Storage for 1 hour (upper graph) and 6 hours (middle graph). The lower graph shows the lack of hemolysis caused by reducing the sedimentation rate by centrifugation after 7 hours of storage.

[0045] Figure 3Graph showing the sedimentation rate of erythrocytes after addition of sucralose in powder form to the indicated final concentrations. Samples were stored for 1 hour (upper graph) and 6 hours (lower graph). NF = No formulation control.

[0046] Figure 4 The effect of different anticoagulants on the sedimentation rate of erythrocytes collected from whole blood is shown. Samples were collected into blood collection tubes containing the indicated anticoagulants. After adding 0.5 M sucralose solution or PBS, samples were stored for 2 hours (upper graph) and 8 hours (lower graph). NF = No formulation control.

[0047] Figure 5 Graph showing the effect of 0.5M sucralose solution on the sedimentation rate of erythrocytes in whole blood compared to PBS, the indicated polyols and the indicated halogenated polyols. Samples were stored for 2 hours. NF = No formulation control. DETAILED DESCRIPTION

[0048] The present invention relates to formulations, compositions, articles of manufacture, kits and methods for reducing the sedimentation rate of one or more red blood cells in a blood sample.

[0049] In some embodiments, the formulations, methods and compositions provided herein provide a reduced sedimentation rate and, therefore, storage of one or more red blood cells in a blood sample at an injection site in a microfluidic device. Reducing the sedimentation rate of one or more red blood cells in a blood sample allows one or more red blood cells to be slowly injected into a microinjection device without prior sample mixing.

[0050] Erythrocyte sedimentation rate is used as a parameter for the prognosis of diseases such as multiple myeloma, temporal arteritis, polymyalgia rheumatica, systemic lupus erythematosus, and rheumatoid arthritis. Therefore, in some embodiments, the formulations, methods, and compositions provided herein for reducing the erythrocyte sedimentation rate in a blood sample can benefit patients suffering from diseases associated with increased erythrocyte sedimentation rate.

[0051] definition

[0052] As used in this specification and the appended claims, the singular forms "a," "an," and "the" include plural referents unless the context clearly dictates otherwise. Thus, for example, reference to "the method" includes one or more methods and / or steps of the type described herein that will become apparent to those skilled in the art upon reading this disclosure and so forth.

[0053] As used herein, "about" in reference to a measurable value such as an amount, a time interval, etc., is intended to include deviations of ±20% or ±10% or ±5% or even ±1% from the specified value, as such deviations are appropriate for the disclosed compositions or practice of the disclosed methods.

[0054] Unless defined otherwise, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention belongs.

[0055] As used herein, the term "ambient temperature" refers to normal room temperature. In some embodiments, the ambient temperature is 15°C to 32°C. In some embodiments, the ambient temperature is 20°C to 27°C.

[0056] As used herein, the terms "reduced sedimentation rate", "reduced sedimentation rate" and "reduction in sedimentation rate" refer to the ability of a material to reduce the sedimentation rate of red blood cells in a blood sample. In some embodiments, the red blood cell sedimentation rate is reduced by at least 10%, at least 20%, at least 30%, at least 40%, at least 50%, at least 60%, at least 70%, at least 80%, at least 90%, at least 95% of the red blood cell sedimentation rate of an untreated blood sample. In some embodiments, the reduction in sedimentation rate refers to the ability of a material to prevent one or more red blood cells in a blood sample from settling from suspension due to gravity. In some embodiments, one or more red blood cells remain suspended for at least 30 minutes. In some embodiments, at least 10%, at least 20%, at least 30%, at least 40%, at least 50%, at least 60%, at least 70%, at least 80%, at least 90%, at least 95% of one or more red blood cells in a treated blood sample remain suspended compared to an untreated blood sample. In some embodiments, the one or more red blood cells remain suspended for at least 10 minutes, at least 20 minutes, at least 30 minutes, at least 40 minutes, at least 50 minutes, at least 60 minutes, at least 90 minutes, at least 2 hours, at least 3 hours, at least 4 hours, at least 5 hours, at least 6 hours, at least 7 hours, at least 8 hours, at least 10 hours, at least 12 hours, at least 24 hours, or at least 48 hours.

[0057] Preparation of reagents

[0058] pH Buffers

[0059] According to certain embodiments, the formulations and compositions described herein for reducing the sedimentation rate of one or more red blood cells in a blood sample include one or more pH buffers. In some embodiments, the pH buffer is any of a large number of compounds known in the art for its ability to resist pH changes in a solution (such as in an aqueous solution in which a pH buffer is present). The selection of one or more specific pH buffers for inclusion in a stable storage composition can be based on the present disclosure and according to conventional practice in the art, and can be affected by a variety of factors, including the pH to be maintained, the nature of the biological sample, the solvent conditions to be used, other components of the formulation to be used, and other conditions. For example, in general, a pH buffer is selected with a proton dissociation constant (pK a ), the proton dissociation constant being a characteristic of the buffer.

[0060] Non-limiting examples of pH buffers include citric acid, tartaric acid, malic acid, sulfosalicylic acid, sulfoisophthalic acid, oxalic acid, borates, CAPS (3-(cyclohexylamino)-1-propanesulfonic acid), CAPSO (3-(cyclohexylamino)-2-hydroxy-1-propanesulfonic acid), EPPS (4-(2-hydroxyethyl)-1-piperazinepropanesulfonic acid), HEPES (4-(2-hydroxyethyl)piperazine-1-ethanesulfonic acid), MES (2-(N-morpholino)ethanesulfonic acid), MOPS (3-(N-morpholino)propanesulfonic acid), MOPSO (3-morpholino-2-hydroxypropanesulfonic acid), PIPE S (1,4-piperazine diethanesulfonic acid), TAPS (N-[tris(hydroxymethyl)methyl]-3-aminopropanesulfonic acid), TAPSO (2-hydroxy-3-[tris(hydroxymethyl)methylamino]-1-propanesulfonic acid), TES (N-[tris(hydroxymethyl)methyl]-2-aminoethanesulfonic acid), bicine (N,N-bis(2-hydroxyethyl)glycine), tricine (N-[tris(hydroxymethyl)methyl]glycine), tris (tris(hydroxymethyl)aminomethane) and bis-tris (2-[bis(2-hydroxyethyl)amino]-2-(hydroxymethyl)-1,3-propanediol). In some embodiments, the formulation has a pH of about 4.0, 4.1, 4.2, 4.3, 4.4, 4.5, 4.6, 4.7, 4.8, 4.9, 5.0, 5.1, 5.2, 5.3, 5.4, 5.5, 5.6, 5.7, 5.8, 5.9, 6.0, 6.1, 6.2, 6.3, 6.4, 6.5, 6.6, 6.7, 6.8, 6.9, 7.0, 7.1, 7.2, 7.3, 7.4, 7.5, 7.6, 7.7, 7.8, 7.9, 8.0, 8.1, 8.2, 8.3, 8.4, 8.5, 8.6, 8.7, 8.8, 8.9, or 9.0.

[0061] Disaccharide derivatives

[0062] In certain embodiments, the preparation or composition for reducing the sedimentation rate of red blood cells in a blood sample comprises at least one halogenated disaccharide derivative. In some embodiments, the halogenated disaccharide derivative is a dichlorinated disaccharide or a trichlorinated disaccharide. In some embodiments, unexpectedly, such dichlorinated disaccharides or trichlorinated disaccharides are able to reduce the sedimentation rate of red blood cells in a blood sample alone or in the presence of only a buffer. Halogenated disaccharide derivatives are known, for example, see U.S. Patent Publication No. 2014 / 0065062, and include sucralose (1,6-dichloro-1,6-dideoxy-β-D-fructofuranosyl-4-chloro-4-deoxy-α-D-galactopyranoside), maltose trichloride, 1,6-dichloro-1,6-dideoxy-β-D-fructofuranosyl-4-chloro-4-deoxy-6-O-monododecanoate-α-D-galactopyranoside, and 1,6-dichloro-1,6-dideoxy-β-D-fructofuranosyl-4-chloro-4-deoxy-6-O-monotetradecanoate-α-D-galactopyranoside. The selection of one or more specific halogenated disaccharide derivatives for inclusion in a formulation or composition for reducing the sedimentation rate of erythrocytes in a blood sample can be made based on this disclosure and in accordance with routine practice in the art, and may be influenced by a variety of factors including other formulation components.

[0063] In some embodiments, the halogenated disaccharide derivative is sucralose. In some embodiments, the sucralose is provided as a solution for mixing with a blood sample. In some embodiments, the solution is an aqueous solution. In some embodiments, the sucralose is present in the formulation at about 5-500mM. In some embodiments, the sucralose is present in the formulation at about 10-500mM. In some embodiments, the sucralose is present in the formulation at about 50-500mM. In some embodiments, the sucralose is present in the formulation at about 100-500mM. In some embodiments, the sucralose is present in the formulation at about 250-500mM. In some embodiments, the sucralose is present in the formulation at about 5-630mM. In some embodiments, the sucralose is present in the formulation at about 5-750mM. In some embodiments, the sucralose is present in the formulation at about 10-750mM. In some embodiments, the sucralose is present in the formulation at about 50-750 mM. In some embodiments, the sucralose is present in the formulation at about 100-750 mM. In some embodiments, the sucralose is present in the formulation at about 250-750 mM. In some embodiments, the formulation is a mixture of water and sucralose.

[0064] In some embodiments, the preparation is provided in an amount sufficient to produce a final concentration of sucralose of about 5 to about 25 mM when mixed with a blood sample. In some embodiments, the sucralose is present in the preparation at about 500 mM and is mixed with the blood sample at a ratio of 1: 20 (v / v) (preparation to blood). In some embodiments, the sucralose is present in the preparation at greater than 25 mM up to 100 mM. In some embodiments, the sucralose is present in the preparation at about 13-24 mM. In some embodiments, the sucralose is provided in powder form as a preparation for mixing with a blood sample. In some embodiments, the sucralose powder is provided in an amount sufficient to produce a final concentration of sucralose of about 5 to about 25 mM when mixed with a blood sample.

[0065] In some embodiments, when mixed with a blood sample, the sucralose is present at a final concentration of about 5-100mM. In some embodiments, when mixed with a blood sample, the sucralose is present at a final concentration of about 5-50mM. In some embodiments, when mixed with a blood sample, the sucralose is present at a final concentration of about 5-25mM. In some embodiments, when mixed with a blood sample, the sucralose is present at a final concentration of about 5mM up to but not including 25mM. In some embodiments, when mixed with a blood sample, the sucralose is present at a final concentration of about 5-20mM. In some embodiments, when mixed with a blood sample, the sucralose is present at a final concentration of about 5-15mM. In some embodiments, when mixed with a blood sample, the sucralose is present at a final concentration of about 10-20mM. In some embodiments, when mixed with a blood sample, the sucralose is present at a final concentration of about 10-15mM. In some embodiments, the sucralose is present at a final concentration of about 5-10 mM when mixed with a blood sample. In some embodiments, the sucralose is present at a final concentration of about 25 mM when mixed with a blood sample.

[0066] Anticoagulants

[0067] In some embodiments, an anticoagulant is included in the formulations and compositions described herein. Such anticoagulants are known in the art. Exemplary anticoagulants include acid citrate dextrose solution (ACD), ethylenediaminetetraacetic acid (EDTA), tripotassium ethylenediaminetetraacetate (K 3 EDTA), ethylenediaminetetraacetic acid dipotassium (K 2 In some embodiments, the anticoagulant is contained in a blood collection tube.

[0068] Exemplary formulations for reducing erythrocyte sedimentation rate in a blood sample

[0069] In some embodiments, formulations comprising sucralose are described herein. In some embodiments, when mixed with a blood sample, the sucralose is present at a final concentration of about 5mM sucralose to about 50mM sucralose. In some embodiments, when mixed with a blood sample, the sucralose is present at a final concentration of about 5mM sucralose to about 25mM sucralose. In some embodiments, the sucralose is present at a final concentration of about 5mM sucralose up to but not including 25mM sucralose. In some embodiments, the sucralose is present at a final concentration of about 5mM sucralose to about 20mM sucralose. In some embodiments, the sucralose is present at a final concentration of about 5mM sucralose to about 15mM sucralose. In some embodiments, the sucralose is present at a final concentration of about 5mM sucralose to about 10mM sucralose. In some embodiments, the sucralose is present as a solution. In some embodiments, the solution is an aqueous solution. In some embodiments, the sucralose is present as a powder. In some embodiments, the formulation further comprises an anticoagulant. In some embodiments, the anticoagulant is selected from acid citrate dextrose solution (ACD), sodium heparin, sodium fluoride, lithium heparin, tripotassium ethylenediaminetetraacetate (K 3 EDTA), ethylenediaminetetraacetic acid dipotassium (K 2 In some embodiments, the anticoagulant is an acid citrate dextrose solution (ACD). In some embodiments, the anticoagulant is sodium heparin. In some embodiments, the anticoagulant is sodium fluoride. In some embodiments, the anticoagulant is lithium heparin. In some embodiments, the anticoagulant is tripotassium ethylenediaminetetraacetate (K 3 In some embodiments, the anticoagulant is dipotassium ethylenediaminetetraacetate (K 2 In some embodiments, the anticoagulant is hirudin. In some embodiments, the anticoagulant is sodium polyanethol sulfonate (SPS). In some embodiments, the preparation is contained in a blood collection tube.

[0070] Method for preparing a preparation for reducing the erythrocyte sedimentation rate in a blood sample

[0071] The methods for preparing the formulations described herein for reducing the erythrocyte sedimentation rate in a blood sample employ techniques known to those skilled in the art and typically use commercially available reagents. In some embodiments, the formulation is prepared as a concentrated (e.g., 2x, 5x, 10x, 20x concentrated, etc.) stock solution of the formulation reagent to be mixed with the blood sample in an appropriate ratio to produce the desired final concentration of sucralose in the blood sample.

[0072] Composition of red blood cells with reduced sedimentation rate in a blood sample

[0073] In some embodiments, described herein are compositions comprising a blood sample and sucralose, wherein the concentration of the sucralose is about 5 mM sucralose up to but not including about 25 mM sucralose. In some embodiments, the composition further comprises an anticoagulant. In some embodiments, the anticoagulant is selected from acid citrate dextrose solution (ACD), sodium heparin, sodium fluoride, lithium heparin, tripotassium ethylenediaminetetraacetate (K 3 EDTA), ethylenediaminetetraacetic acid dipotassium (K 2 In some embodiments, the anticoagulant is an acid citrate dextrose solution (ACD). In some embodiments, the anticoagulant is an acid citrate dextrose solution A (ACD-A). In some embodiments, the anticoagulant is an acid citrate dextrose solution B (ACD-B). In some embodiments, the anticoagulant is sodium heparin. In some embodiments, the anticoagulant is sodium fluoride. In some embodiments, the anticoagulant is lithium heparin. In some embodiments, the anticoagulant is tripotassium ethylenediaminetetraacetate (K 3 In some embodiments, the anticoagulant is dipotassium ethylenediaminetetraacetate (K 2 In some embodiments, the anticoagulant is hirudin. In some embodiments, the anticoagulant is sodium polyanethol sulfonate (SPS). In some embodiments, the composition is contained in a blood collection tube. In some embodiments, the blood collection tube is an evacuated blood collection tube. In some embodiments, the blood is collected from a subject. In some embodiments, the subject is a mammal. In some embodiments, the subject is a human. In some embodiments, the composition of one or more red blood cells with reduced sedimentation rate in a blood sample is stored for an extended period of time in a formulation described herein, such as in a microfluidic device, prior to analysis.

[0074] Products

[0075] In certain embodiments, a product is provided, which comprises a preparation provided herein, which is contained in a suitable blood collection tube, a blood collection container or a blood collection vessel for collecting a biological sample. In some embodiments, these products are used to reduce the sedimentation rate of one or more red blood cells in a blood sample when a biological sample is collected. In certain embodiments, the blood collection tube is an evacuated blood collection tube with an air pressure lower than atmospheric pressure, for extracting a predetermined volume of whole blood. In some embodiments, the blood collection tube contains about 28.6 mg of sucralose powder, and the size of the blood collection tube is designed to accommodate a blood draw volume of 3.0 mL of blood, thereby producing a final sucralose concentration of about 24 mM after adding 3.0 mL of blood. In some embodiments, the blood collection tube contains about 33.4 mg of sucralose powder, and the size of the blood collection tube is designed to accommodate a blood draw volume of 3.5 mL of blood. In some embodiments, the blood collection tube contains about 42.9 mg of sucralose powder, and the size of the blood collection tube is designed to accommodate a blood draw volume of 4.5 mL of blood. In some embodiments, the blood collection tube contains about 52.4 mg of sucralose powder, and the size of the blood collection tube is designed to accommodate a blood draw volume of 5.5 mL of blood. In some embodiments, the blood collection tube contains about 95.4 mg of sucralose powder, and the size of the blood collection tube is designed to accommodate a blood draw volume of 10 mL of blood. In some embodiments, these articles are used in the kits and methods described herein.

[0076] Reagent test kit

[0077] In certain embodiments, a kit comprising any of the articles described herein and a package insert is provided. In some embodiments, the components of the kit are provided in a container. In some embodiments, the container is a plastic enclosure with compartments. In some embodiments, the container includes a tightly sealable lid so that the contents of the kit can be sterilized and sealed for storage.

[0078] Method for reducing the sedimentation rate of red blood cells in a blood sample

[0079] In some embodiments, described herein is an in vitro method for reducing the erythrocyte sedimentation rate in a blood sample, the method comprising: combining a blood sample with an amount of a formulation comprising sucralose, wherein the amount is sufficient to produce a treated blood sample having a sucralose concentration of at least about 5 mM sucralose, thereby reducing the erythrocyte sedimentation rate compared to the erythrocyte sedimentation rate in an untreated blood sample. In some embodiments, the treated blood sample has a sucralose concentration of about 5 mM sucralose to about 50 mM sucralose. In some embodiments, the treated blood sample has a sucralose concentration of about 5 mM sucralose to about 25 mM sucralose. In some embodiments, the treated blood sample has a sucralose concentration of about 5 mM sucralose up to but not including 25 mM sucralose. In some embodiments, the erythrocyte sedimentation rate is reduced by at least 10%, at least 20%, at least 30%, at least 40%, at least 50%, at least 60%, at least 70%, at least 80%, at least 90%, at least 95% of the erythrocyte sedimentation rate of an untreated blood sample. In some embodiments, the formulation is a powder. In some embodiments, the formulation is a solution. In some embodiments, the solution is an aqueous solution. In some embodiments, the formulation consists of sucralose. In some embodiments, the formulation further comprises an anticoagulant. In some embodiments, the anticoagulant is selected from acid citrate dextrose solution (ACD), sodium heparin, sodium fluoride, lithium heparin, tripotassium ethylenediaminetetraacetate (K 3 EDTA), ethylenediaminetetraacetic acid dipotassium (K 2 In some embodiments, the anticoagulant is an acid citrate dextrose solution (ACD). In some embodiments, the anticoagulant is sodium heparin. In some embodiments, the anticoagulant is sodium fluoride. In some embodiments, the anticoagulant is lithium heparin. In some embodiments, the anticoagulant is tripotassium ethylenediaminetetraacetate (K 3 In some embodiments, the anticoagulant is dipotassium ethylenediaminetetraacetate (K 2 In some embodiments, the anticoagulant is hirudin. In some embodiments, the anticoagulant is sodium polyanethol sulfonate (SPS). In some embodiments, the preparation is contained in a blood collection tube, and the combining step occurs in the blood collection tube. In some embodiments, the blood collection tube is an evacuated blood collection tube. In some embodiments, the blood is collected from a subject. In some embodiments, the subject is a mammal. In some embodiments, the subject is a human.

[0080] In some embodiments, the present invention describes a method for keeping one or more red blood cells suspended in a blood sample, the method comprising: combining a blood sample with a certain amount of a preparation comprising sucralose, wherein the amount is sufficient to produce a treated blood sample having a sucralose concentration of at least about 5mM sucralose, whereby the one or more red blood cells remain suspended for at least 30 minutes compared to an untreated blood sample. In some embodiments, the one or more red blood cells remain suspended for at least 10 minutes, at least 20 minutes, at least 30 minutes, at least 40 minutes, at least 50 minutes, at least 1 hour, at least 2 hours, at least 3 hours, at least 4 hours, at least 5 hours, at least 6 hours, at least 7 hours, at least 8 hours, at least 10 hours, at least 12 hours, at least 24 hours, or at least 48 hours. In some embodiments, the treated blood sample has a sucralose concentration of about 5mM sucralose to about 25mM sucralose. In some embodiments, the treated blood sample has a sucralose concentration of about 5mM sucralose up to but not including 25mM sucralose. In some embodiments, at least 10%, at least 20%, at least 30%, at least 40%, at least 50%, at least 60%, at least 70%, at least 80%, at least 90%, at least 95% of the one or more red blood cells remain suspended in the treated blood sample compared to the untreated blood sample. In some embodiments, the formulation is a powder. In some embodiments, the formulation is a solution. In some embodiments, the solution is an aqueous solution. In some embodiments, the formulation consists of sucralose. In some embodiments, the formulation further comprises an anticoagulant. In some embodiments, the anticoagulant is selected from acid citrate dextrose solution (ACD), sodium heparin, sodium fluoride, lithium heparin, tripotassium ethylenediaminetetraacetate (K 3 EDTA), ethylenediaminetetraacetic acid dipotassium (K 2 In some embodiments, the anticoagulant is an acid citrate dextrose solution (ACD). In some embodiments, the anticoagulant is sodium heparin. In some embodiments, the anticoagulant is sodium fluoride. In some embodiments, the anticoagulant is lithium heparin. In some embodiments, the anticoagulant is tripotassium ethylenediaminetetraacetate (K 3 In some embodiments, the anticoagulant is dipotassium ethylenediaminetetraacetate (K 2In some embodiments, the anticoagulant is hirudin. In some embodiments, the anticoagulant is sodium polyanethol sulfonate (SPS). In some embodiments, the preparation is contained in a blood collection tube, and the combining step occurs in the blood collection tube. In some embodiments, the blood collection tube is an evacuated blood collection tube. In some embodiments, the blood sample is collected from a subject. In some embodiments, the subject is a mammal. In some embodiments, the subject is a human.

[0081] Blood collection tubes, blood collection bags, blood collection containers and blood collection vessels are well known in the art and have been used by practitioners for decades. Blood collected for the purpose of reducing the erythrocyte sedimentation rate can be obtained using any method or apparatus commonly used by those skilled in the art, such as venipuncture or fingerstick. In some embodiments, when blood is collected by venipuncture, when the blood sample is obtained from the subject, the preparation is located in a blood collection tube, for example, a vacuum tube ( Blood collection tubes, Becton Dickinson or In some embodiments, when blood is collected by venipuncture, the preparation is added to the obtained whole blood sample immediately or shortly after the blood is drawn.

[0082] In some embodiments, the methods described herein employ the disclosed articles and kits.

[0083] The following examples are given by way of illustration and not limitation.

[0084] Example 1

[0085] Reduction of erythrocyte sedimentation rate in whole blood by addition of sucralose

[0086] This example describes the reduction of erythrocyte sedimentation in whole blood by the addition of sucralose.

[0087] Fresh blood was collected from BD K 2 EDTA tubes and combined. Blood was mixed with 0.5M sucralose solution at a ratio of 20:1 by aliquoting 952 μL of fresh blood into 2 mL centrifuge tubes containing 48 μL of formulation to obtain a concentration of about 24 mM sucralose. 48 μL of PBS and 0.9% saline were used as controls. Blood without formulation (NF) was used as an additional control. The filled centrifuge tubes were gently inverted five times to mix and stored upright on a bench at about 25°C. At 0 hours ( Figure 1A ), 6 hours( Figure 1B ) and 24 hours ( Figure 1C ) were taken against a white background for visual analysis of sedimentation rate.

[0088] like Figure 1A -C, compared with whole blood alone (NF) or whole blood supplemented with PBS or saline, K 2 Whole blood collected in EDTA tubes had lower erythrocyte sedimentation rates when stored in aliquots at ambient temperature for 6 and 24 hours.

[0089] Example 2

[0090] Effects of sucralose concentration and other sugars on reducing erythrocyte sedimentation rate

[0091] This example illustrates the effect of sucralose concentration and other sugars on reducing erythrocyte sedimentation rate in whole blood.

[0092] Fresh blood was collected from BD K 2 EDTA 952 μL of fresh blood was aliquoted into 2 mL centrifuge tubes, each containing 48 μL of Figure 2 Sucralose at the indicated concentrations, or the indicated sugars (ML848, dichlorinated monosaccharides, or DG783, monofluorinated monosaccharides) at the indicated concentrations. Sucralose solutions of varying concentrations were prepared by diluting 0.5M sucralose to 10 mM in water. The formulations were adjusted to 300 mOsmol with NaCl, except for the highest concentration formulation which had an osmotic pressure of 500 mOsmol. 48 μL of PBS and 0.9% saline were used as controls. Figure 2 The final concentrations of sucralose or the indicated sugars shown in the figures are as follows (from left to right): 24 mM sucralose, 12 mM sucralose, 6.24 mM sucralose, 2.88 mM sucralose, 1.44 mM sucralose, 0.96 mM sucralose, 0.48 mM sucralose, 24 mM sucrose, 2.4 mM sucrose, 24 mM trehalose, 24 mM melezitose, 4.8 mM ML848, and 24 mM DG783.

[0093] The filled centrifuge tube was gently inverted five times to mix and stored upright on the bench at approximately 25°C. Figure 2 , above) and 6 hours ( Figure 2 , middle panel) against a white background for visual analysis of sedimentation rate. After 7 hours of storage, the tubes were centrifuged at 3000 rpm for 20 min to determine the effect of hemolysis on sedimentation rate by visual analysis ( Figure 2 , figure below).

[0094] Figure 2 Shown are the K values ​​of cells containing high concentrations of sucralose after aliquots were stored at ambient temperature for 1 hour (upper panel) and 6 hours (middle panel).2 Whole blood collected in EDTA tubes had a lower erythrocyte sedimentation rate. No lower erythrocyte sedimentation rate was observed in samples with a final sucralose concentration below 5 mM, or in samples after the addition of sucrose, trehalose, melezitose, ML848, DG783, PBS, or saline to whole blood.

[0095] The homogeneity of whole blood in the presence of high concentrations of sucralose was not due to excessive hemolysis of erythrocytes, as shown by centrifugation of samples after 7 hours of storage ( Figure 2 , lower panel). Centrifugation resulted in a clear separation of plasma without coloration of the plasma layer, similar to that observed for saline and PBS sample controls ( Figure 2 , lower panel). No significant hemolysis was observed at final sucralose concentrations of 40 mM or below.

[0096] Example 3

[0097] Effect of sucralose added as a powder on reducing erythrocyte sedimentation rate

[0098] This example describes the effect of adding sucralose in powder form on reducing the erythrocyte sedimentation rate in whole blood.

[0099] Fresh blood was collected from BD K 2 EDTA Tubes and combined. Aliquot 1 mL of fresh blood into 2 mL centrifuge tubes containing sucralose powder. The final concentration of sucralose ranged from 7.5 mM to 100 mM, corresponding to 3.0 mg to 39.8 mg per mL of blood. No formulation (NF) control samples did not receive the addition of sucralose. Invert the tubes at least five times until there is no visible undissolved material at the bottom of the tube. Store the tubes upright on a benchtop at approximately 25°C. After 1 hour ( Figure 3 , above) and 6 hours ( Figure 3 , lower panel) the tubes were photographed against a white background for visual analysis of sedimentation rate.

[0100] Figure 3 The data in show that the rate of erythrocyte sedimentation is inversely proportional to the concentration of sucralose. At sucralose concentrations of 24 mM or higher, homogeneity of whole blood samples stored at ambient temperature for 6 hours was observed, while lower concentrations of sucralose resulted in separation of erythrocytes from the plasma layer.

[0101] Example 4

[0102] Effect of blood collection conditions on reducing erythrocyte sedimentation rate

[0103] This example describes the effect of different anticoagulants present at the time of blood collection on reducing the erythrocyte sedimentation rate in whole blood.

[0104] To screen the collection conditions, fresh blood was collected into a series of BD containing different anticoagulants. or Greiner Bio-One In small volume blood collection tubes, the anticoagulants include acid citrate dextrose solution B (ACD-B), tripotassium ethylenediaminetetraacetate (K 3 EDTA), sodium heparin (NaHep), lithium heparin (LiHep), sodium fluoride (NaF) and sodium polyanethol sulfonate (SPS). 2 As a control, 952 μL of fresh blood was aliquoted into 2 mL centrifuge tubes containing 48 μL of 0.5 M sucralose or PBS. No formulation (NF) was added as an additional control. The filled centrifuge tubes were gently inverted five times to mix and stored upright on a bench at approximately 25°C. Figure 4 , above) and 8 hours ( Figure 4 , lower figure) the tubes were photographed against a white background for visual analysis of sedimentation rate. Figure 4 As shown, for each anticoagulant present in the blood collection tube at the time of blood collection, the addition of 0.5 M sucralose up to a final sucralose concentration of 24 mM in the treated sample resulted in a decrease in erythrocyte sedimentation rate. In contrast, the addition of PBS had no effect on erythrocyte sedimentation rate.

[0105] Example 5

[0106] Effect of sucralose compared with polyols and halogenated polyols on reducing erythrocyte sedimentation rate

[0107] This example describes the effect of sucralose on reducing erythrocyte sedimentation rate in whole blood compared to polyols and halogenated polyols.

[0108] Fresh blood was collected from BD K 2 EDTA Tube and merge. 952 μL fresh blood was aliquoted into 2 mL centrifuge tubes containing 48 μL of 0.5M sucralose, a specified polyol or a specified halogenated polyol solution. 0.25M additive was adjusted to 300 mOsmol with NaCl. PBS was added without formulation (NF) as a control. The filled centrifuge tube was gently inverted five times for mixing and stored upright on a table at approximately 25°C. The tube was photographed relative to a white background at 2 hours for visual analysis of sedimentation rate.

[0109] Figure 5The data in show that the addition of 0.5 M sucralose to a final sucralose concentration of 24 mM in the treated samples resulted in a decrease in the erythrocyte sedimentation rate. In contrast, the addition of the indicated polyols or halogenated polyols had no significant effect on erythrocyte sedimentation.

[0110] Unless the context requires otherwise, throughout this specification and claims, the word "comprising" and its variations, which are used interchangeably with "including," "containing," or "characterized by," are inclusive or open-ended language and do not exclude additional, unlisted elements or method steps. The phrase "consisting of" excludes any element, step, or ingredient not specified in the claim. The phrase "consisting essentially of" limits the scope of the claim to the specified materials or steps and those that do not substantially affect the basic and novel characteristics of the claimed invention. The present disclosure encompasses embodiments of the compositions and methods of the present invention that correspond to the scope of each of these phrases. Therefore, a composition or method comprising the recited elements or steps encompasses specific embodiments in which the composition or method consists essentially of or consists of these elements or steps.

[0111] Reference throughout this specification to "one embodiment" or "an embodiment" or "an aspect" means that the specific features, structures, or characteristics described in connection with the embodiment are included in at least one embodiment of the present invention. Thus, the appearance of the phrase "in one embodiment" or "in an embodiment" throughout this specification does not necessarily refer to the same embodiment. Furthermore, the specific features, structures, or characteristics may be combined in any suitable manner in one or more embodiments.

[0112] The various embodiments described above may be combined to provide further embodiments. These and other changes may be made to the embodiments in accordance with the description detailed above. In summary, in the appended claims, the terms used should not be interpreted as limiting the claims to the specific embodiments disclosed in the specification and claims, but should be interpreted as including all possible embodiments and the full range of equivalents for which the claims are requested. Therefore, the claims are not limited by the disclosure.

[0113] Although preferred embodiments of the present invention have been shown and described herein, it will be apparent to those skilled in the art that these embodiments are provided by way of example only. Those skilled in the art will now appreciate multiple variations, changes and substitutions without departing from the present invention. It should be understood that various alternatives to the embodiments of the present invention described herein can be used to implement the present invention. It is intended to define the scope of the present invention with the following claims, and thus encompass methods and structures and their equivalents within the scope of these claims.

Claims

1. A method for analyzing a blood sample, the method comprising: injecting the processed blood sample into the microfluidic device; and analyzing the processed blood sample; wherein the treated blood sample comprises a blood sample from a subject and a formulation comprising 5 mM to 100 mM sucralose, wherein the treated blood sample comprises a reduced erythrocyte sedimentation rate compared to an untreated blood sample.

2. The method of claim 1, wherein the processed blood sample is stored prior to injection into the microfluidic device.

3. The method of claim 2, wherein the processed blood sample is stored for 10 minutes to 48 hours.

4. The method of claim 2, wherein the processed blood sample is stored for 30 minutes to 24 hours. The method according to claim 2 , wherein the processed blood sample is stored for 1 hour to 12 hours.

6. The method of claim 2, wherein the processed blood sample is stored at ambient temperature.

7. The method of claim 2, wherein the processed blood sample is not mixed prior to injection into the microfluidic device.

8. The method of claim 1, wherein the erythrocyte sedimentation rate is reduced by at least 10% compared to the erythrocyte sedimentation rate of an untreated blood sample.

9. The method of claim 1, wherein the erythrocyte sedimentation rate is reduced by at least 25% compared to the erythrocyte sedimentation rate of an untreated blood sample.

10. The method of claim 1, wherein the erythrocyte sedimentation rate is reduced by at least 50% compared to the erythrocyte sedimentation rate of an untreated blood sample.

11. The method of claim 1, wherein the subject is a human subject.

12. The method of claim 11, wherein the human subject is diagnosed with a disease associated with increased erythrocyte sedimentation rate.

13. The method of claim 12, wherein the disease is selected from the group consisting of multiple myeloma, temporal arteritis, polymyalgia rheumatica, systemic lupus erythematosus, and rheumatoid arthritis.

14. The method of claim 1, wherein the formulation comprises 5 mM to 50 mM sucralose by adding a blood sample of the subject.

15. The method of claim 1, wherein the formulation comprises 5 mM to 25 mM sucralose by adding a blood sample of the subject.

16. The method of claim 1, wherein the formulation further comprises an anticoagulant.

17. The method of claim 16, wherein the anticoagulant is selected from the group consisting of acid citrate dextrose solution, sodium heparin, sodium fluoride, lithium heparin, tripotassium ethylenediaminetetraacetate, dipotassium ethylenediaminetetraacetate, hirudin, and sodium polyanethol sulfonate.

18. The method of claim 1, wherein the treated blood sample comprises at least 2.0 mg of the sucralose per ml of the subject's blood sample.

19. The method of claim 1, wherein the formulation is in a lyophilized form or in the form of an aqueous solution, and wherein the formulation is contained in a blood collection tube.

20. A blood collection tube comprising a preparation for reducing the sedimentation rate of erythrocytes, the preparation comprising an anticoagulant and sucralose.

21. The blood collection tube of claim 20, wherein the sucralose is present in an amount sufficient to provide a final concentration of 5.0 mM to 100 mM sucralose upon addition of a blood sample.

22. The blood collection tube of claim 20, wherein the sucralose is present in an amount sufficient to provide a final concentration of 5.0 mM to 50 mM sucralose upon addition of a blood sample.

23. The blood collection tube of claim 20, wherein the sucralose is present in an amount sufficient to provide a final concentration of 5.0 mM to 25 mM sucralose upon addition of a blood sample.

24. The blood collection tube according to claim 20, wherein the anticoagulant is selected from the group consisting of acid citrate dextrose solution, sodium heparin, sodium fluoride, lithium heparin, tripotassium ethylenediaminetetraacetate, dipotassium ethylenediaminetetraacetate, hirudin and sodium polyanethole sulfonate.

25. The blood collection tube according to claim 20, wherein the preparation is in a lyophilized form or in the form of an aqueous solution.

26. The blood collection tube of claim 20, wherein the tube is a vacuum blood tube.

27. The blood collection tube according to claim 20, wherein the formulation consists of an anticoagulant and sucralose.

28. The blood collection tube of claim 20, wherein the formulation reduces the erythrocyte sedimentation rate of the treated blood sample by at least 10% compared to the erythrocyte sedimentation rate of the untreated blood sample.

29. The blood collection tube of claim 20, wherein the formulation reduces the erythrocyte sedimentation rate of the treated blood sample by at least 25% compared to the erythrocyte sedimentation rate of the untreated blood sample.

30. The blood collection tube of claim 20, wherein the formulation reduces the erythrocyte sedimentation rate of the treated blood sample by at least 50% compared to the erythrocyte sedimentation rate of the untreated blood sample.

31. The blood collection tube of claim 20, wherein the formulation reduces the erythrocyte sedimentation rate of the treated blood sample when compared to the erythrocyte sedimentation rate of an untreated blood sample during storage for at least 24 hours.

32. The blood collection tube of claim 20, wherein the formulation reduces the erythrocyte sedimentation rate of the treated blood sample when compared to the erythrocyte sedimentation rate of an untreated blood sample during storage for at least 12 hours.

33. The blood collection tube of claim 20, wherein the formulation reduces the erythrocyte sedimentation rate of the treated blood sample when compared to the erythrocyte sedimentation rate of an untreated blood sample during storage for at least 6 hours.

34. The blood collection tube of claim 20, wherein the formulation reduces the erythrocyte sedimentation rate of the treated blood sample during storage at ambient temperature compared to the erythrocyte sedimentation rate of an untreated blood sample.

35. The blood collection tube according to claim 20, wherein at least 2.0 mg of the sucralose is present per ml of blood sample added to the blood collection tube.

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