Blood component collection and separation medium, blood component collection and separation device, and blood component separation and extraction method

Abstract

The present invention relates to a blood component collection and separation medium (1) comprising a substrate (3) having a maximum flow pore diameter that enables at least red blood cells to be trapped on the surface of the substrate (3), the blood component collection and separation medium (1) comprising a boundary wall (7) forming a pattern (9) and made of a hydrophobic resin, the pattern (9) having: a collection zone (91); at least one storage area (93) intended to collect at least one component in the whole blood sample (5); and at least one channel (95) connecting the collection area (91) to the at least one storage area (93), the channel (95) forming a bottleneck between the collection area (91) and the storage area (93). The invention further relates to a blood component collection and separation device and a blood component separation and extraction method.

Description

[0001] This application is a divisional application of the patent application with the application number 201980102257.9, the application date of October 2, 2019, and the invention title of "Blood Component Collection and Separation Medium, Blood Component Collection and Separation Device Comprising the Medium, and Method for Separating and Extracting Blood Components Using the Medium". Technical Field

[0002] The present invention relates to the collection and separation of biological fluids. More specifically, the present invention relates to a blood component collection and separation medium, its use in a method for separating and extracting blood components, and a blood component collection and separation device implementing such a blood component collection and separation medium. More specifically, the present invention relates to a device for use as a blood component collector and separator. Background Art

[0003] Biological samples are often used in laboratory and clinical settings to analyze various components in the sample (e.g., a blood sample). Biological samples are often processed in liquid form. Thus, liquid samples are collected in collection devices, processed, transported to a laboratory, and stored. There are various problems associated with activities around liquid samples, including the risk of container breakage or leakage (which results in sample loss and potential contamination and / or infection risk), sample instability during transportation and storage, and transportation carrier restrictions associated with the transportation of liquid biohazard materials.

[0004] To avoid such problems, various devices and methods for dry samples have emerged. Blood as a dry sample is not considered a biohazard material and can be transported via traditional delivery services like any other package. For this purpose, a blood sample (usually a few drops) is placed on paper and allowed to dry. Additionally, to improve the efficiency of these devices and methods, some papers are configured for separating blood components.

[0005] In fact, blood consists of two parts: blood cells and the liquid in which the blood cells are suspended. This liquid is called blood plasma (the "plasma"), and this liquid is a protein fluid, which may include other smaller cells and dissolved proteins (such as serum albumin, globulin, and fibrinogen), glucose, clotting factors, electrolytes (Na + 、Ca 2+ 、Mg 2+ 、HCO3 - 、Cl -etc.), hormones, and carbon dioxide. The main types of eukaryotic cells present in blood are red blood cells (erythrocytes), white blood cells (leucocytes), and platelets, although other cell types can also be detected. Red blood cells make up approximately half of the volume of a blood sample.

[0006] For rapid diagnosis of plasma components under clinical, hospital, and field conditions, it is very important to separate red blood cells from the plasma in a blood sample because red blood cells can reduce the sensitivity of the analysis. Whole blood can also cause unwanted chemical interference. For example, hemoglobin released from red blood cells can affect the performance of certain clinical assays due to the heme group, which can act as a catalyst.

[0007] During the process of separating red blood cells from the fluid fraction, it is even more important that the red blood cells do not lyse or rupture, which can lead to the release of internal components into the plasma and also contaminate the plasma sample. To obtain plasma from whole blood for testing, four different types of techniques are essentially used: namely, techniques based on gravity, pressure drop, capillary flow, and centrifugal force.

[0008] Assays conducted under field conditions should be inexpensive and the samples should be disposable. Some of the above techniques do not well meet these goals. Pressure drop and centrifugal force require specialized equipment, implementing pumps and centrifuges making them unsuitable for field use. Then, the gravity-based technique is too slow for field use.

[0009] Therefore, assays for separating whole blood into red blood cells and plasma have been developed, which are based on capillary flow in a lateral flow assay device. Many such assays are disclosed in the art.

[0010] U.S. Patent Nos. 4,477,575 and 4,816,224 describe media using glass microfiber layers to separate red blood cells from whole blood. Papers containing 100% of the proposed type of glass microfiber are inherently fragile and require great care during handling. Although the strength can be increased by using adhesives, some adhesives can interfere with the analysis or make the sheet hydrophobic.

[0011] Another method involves using a single-layer medium made of a composite of glass microfibers, cellulose fibers, and synthetic staple fibers. In this regard, reference is made to U.S. Patent No. 5,186,843.

[0012] Although the blood separation media proposed in the past are suitable for their intended purposes, some improvements are still needed.

[0013] Typically, due to the considerable viscosity of whole blood and its tendency to start clotting in air, filtration on a lateral flow assay device is hampered by insufficient velocity. The pores of a standard blood filter must be smaller than the size of red blood cells. This is because red blood cells can fold. When these folded cells pass through the pores, some of the cells remain in the pores, clogging the pores and reducing the filtration efficiency. This also reduces the number of pores available for plasma to pass through and decreases the total amount of plasma passing through the filter per unit time.

[0014] In addition, damage or rupture of the cells may occur. This is especially true if the fibers constituting the nonwoven fabric are glass (such as the glass proposed in U.S. Patent 5,186,843).

[0015] Due to the affinity of red blood cells for glass, glass fibers can still separate red blood cells from blood well, and this affinity provides an effective slowdown of the flow of red blood cells in the capillary. However, even if red blood cells show good affinity for glass fibers, those glass fibers may increase hemolysis, and the separation of multiple blood components may not be so effective. To solve this problem of the prior art, a blood collection and separation medium for preventing such hemolysis is known from the international patent application WO2017 / 017314 filed by the applicant. Hemolysis is prevented by using a resin designed to at least partially coat the glass fibers, and this hemolysis is also prevented. However, even if this blood collection and separation medium provides good separation results for multiple blood components, the deposition of the whole blood sample must be done precisely, and the areas to be perforated or eluted, for example, to recover the desired components in the whole blood sample, may be difficult to identify. In addition, this blood collection and separation medium may be difficult to use in an automated method for collecting and separating blood components from a whole blood sample. Summary of the Invention

[0016] The present invention aims to at least partially solve the disadvantages of the prior art discussed above by providing a blood component collection and separation medium that facilitates the deposition of a whole blood sample.

[0017] Another aspect of the present invention is to provide a blood component collection and separation medium that has easily identifiable areas for the multiple components of a whole blood sample after the multiple components of the whole blood sample are separated on the blood component collection and separation medium, and that prevents hemolysis during the process of separating the multiple components in the whole blood sample.

[0018] Another aspect of the present invention, different from those described above, is to provide a blood component collection and separation device that can be used in an automated method.

[0019] To at least partially meet at least one of the above - mentioned objectives, the present invention relates to a blood - component collection and separation medium, which comprises a substrate intended to be wetted by a whole - blood sample. The substrate has a maximum flow pore size of 8 μm to 35 μm, and such a maximum flow pore size enables at least red blood cells to be retained in the substrate. The blood - component collection and separation medium further comprises boundary walls formed in a pattern in the substrate. The boundary walls are made of a hydrophobic resin, and the pattern has:

[0020] · A collection zone, which is intended to receive a whole - blood sample;

[0021] · At least one storage zone, which is intended to separate at least one component from the whole - blood sample through the substrate

[0022] and store at least one component of the whole - blood sample afterwards; and

[0023] · At least one channel connecting the collection zone to at least one storage zone, and the channel forms a bottleneck between the collection zone and the storage zone.

[0024] The presence of the channel that forms a bottleneck between the collection zone and the storage zone enables plasma to diffuse into at least one storage zone and retains red blood cells in the collection zone. Therefore, the identification of the zones containing specific components of the whole - blood sample can be easily made. In addition, the deposition of the whole - blood sample can be carried out anywhere in the collection zone without affecting the diffusion of plasma into the storage zone, which also simplifies the deposition process of the whole - blood sample on the blood - component collection and separation medium.

[0025] The blood - component collection and separation medium according to the present invention may further have one or more of the following features individually or in combination.

[0026] According to a specific embodiment, the maximum flow pore size of the substrate is preferably 9.5 μm to 10.5 μm.

[0027] The substrate may further have a minimum flow pore size of 0.5 μm to 2 μm, preferably 0.9 μm to 1.3 μm.

[0028] According to a specific embodiment, the substrate may have an average flow pore size of 2.5 μm to 5 μm.

[0029] According to this specific embodiment, the average flow pore size is preferably 3 μm to 3.5 μm.

[0030] The hydrophobic resin forming the boundary walls may be selected from photocurable resins, more preferably UV - curable resins, or thermosetting resins.

[0031] More specifically, the hydrophobic resin may be selected from fluorinated resins, modified fluorinated resins, latexes, glycol ether acrylates, acrylates, or combinations thereof.

[0032] According to a specific embodiment, the hydrophobic resin is selected from polymethyl methacrylate (PMMA) or polydimethylsiloxane (PDMS).

[0033] According to a specific embodiment, the collection area of the pattern may have a substantially parallelogram shape.

[0034] According to this specific embodiment, the at least one storage area may be provided at the corners of the substantially parallelogram shape of the collection area.

[0035] According to a variant of this specific embodiment, the pattern may have as many storage areas as the collection area has corners.

[0036] Optionally or additionally, the at least one storage area may have a substantially circular shape.

[0037] According to a first embodiment, the storage area is intended to be perforated.

[0038] According to a second embodiment, at least one component stored in the storage area is intended to be eluted.

[0039] According to a third embodiment, the collection area is intended to be perforated.

[0040] According to one aspect, the at least one channel has a length of 1 mm to 5 mm and a width of 1 mm to 3 mm.

[0041] According to a specific embodiment, the substrate may be a fibrous web, the fibrous web comprising:

[0042] · A first fiber selected from glass microfibers or synthetic microfibers;

[0043] · A second fiber selected from fibrillated fibers; and

[0044] · A hydrophilic binder.

[0045] The hydrophilic binder may be selected from latex binders.

[0046] According to an alternative, the hydrophilic binder may be selected from polyvinyl alcohol binders.

[0047] According to a further alternative, the hydrophilic binder may be selected from styrene-butadiene binders.

[0048] According to another alternative, the hydrophilic binder may be selected from vinyl acetate binders.

[0049] According to a variant, the hydrophilic binder may be selected from polysaccharide binders.

[0050] According to another variant, the hydrophilic binder can be selected from protein binders.

[0051] According to a specific embodiment, the first fiber can have a diameter of less than 5 μm, and preferably from 0.4 μm to 1 μm.

[0052] Optionally or additionally, the first fiber can have a specific surface area greater than 1.5 m 2 / g.

[0053] Furthermore, the first fiber can have a length / diameter ratio greater than 100, and more preferably greater than 500.

[0054] The first fiber and the second fiber can be mixed together.

[0055] According to a specific embodiment, the second fiber is a cellulose-based fiber, and in particular cotton linter, lyocell fiber or viscose fiber.

[0056] According to the specific embodiment described above, the fibrous web can contain 5 to 7% by weight of a hydrophilic binder.

[0057] According to a specific embodiment, the fibrous web can comprise 50 to 99% by weight, particularly 70 to 98% by weight, of the first fiber, and 1 to 50% by weight of the second fiber, with 100% by weight corresponding to the fibrous web formed.

[0058] The hydrophilic binder can at least partially cover the first fiber of the fibrous web.

[0059] According to one aspect, the fibrous web can comprise at least one additive for improving the properties of the substrate.

[0060] According to an embodiment of this aspect, the at least one additive can be a stabilizer of biomarkers extracted from plasma, such as animal proteins (such as bovine serum albumin (BSA)), gelatin derivatives or sucrose.

[0061] According to a specific embodiment, some or all of the boundary walls extend through at least a part of the depth of the substrate or through the entire depth of the substrate. In particular, the boundary walls preferably extend at least through the entire depth of the substrate.

[0062] According to a specific embodiment, the boundary walls forming the pattern can extend beyond one surface of the substrate.

[0063] The size of the collection area can be designed to receive a whole blood sample with a volume of 50 to 500 μL.

[0064] As an alternative or in addition, the collection zone may further contain a salt capable of effecting at least partial shrinkage of red blood cells, the salt being selected from the group consisting of halides and sulfates of alkali metals and alkaline earth metals, and hydrochlorides of organic bases.

[0065] According to this alternative, the salt may be selected from calcium chloride, potassium sulfate or guanine hydrochloride.

[0066] Other suitable salts are manganese chloride, potassium chloride, magnesium chloride and sodium chloride.

[0067] The salt may be present in the substrate at a concentration of 1-25% by total weight of the substrate, more particularly 10-20% by total weight of the substrate.

[0068] According to a specific embodiment, the pattern may have a collection zone having a square shape and four storage zones provided at each corner of the collection zone, each storage zone having a circular shape and being connected to the collection zone by a channel forming a bottleneck between the collection zone and the associated storage zone.

[0069] The at least one storage zone may have a pre-perforation facilitating its perforation.

[0070] The present invention further relates to a blood component collection and separation device designed to collect and separate at least one component in a whole blood sample, the blood component collection and separation device comprising at least a blood component collection and separation medium as defined above and a frame surrounding the blood component collection and separation medium.

[0071] The blood component collection and separation device according to the present invention may further comprise one or more of the following features.

[0072] According to a specific embodiment, the frame may be made of cardboard.

[0073] The frame may enable the blood component collection and separation device to be picked up and handled by an automaton.

[0074] The blood component collection and separation device may be square or rectangular.

[0075] According to a first specific embodiment, the frame may have a triangle printed on one corner, the triangle being designed to enable an automaton to identify the orientation of the blood component collection and separation device.

[0076] According to a second specific embodiment, the frame may have a notched angle, the notched angle being designed to enable an automaton to identify the orientation of the blood component collection and separation device.

[0077] In addition, the blood component collection and separation device may have a length and width configured to be compatible with an automaton.

[0078] Optionally, the blood component collection and separation device may have at least two indication marks in the collection area, the indication marks being configured to enable an automaton to detect the collection area.

[0079] According to a first variant, the indication marks may be provided on the blood component collection and separation medium outside the pattern.

[0080] According to this first variant, the indication marks may be printed by inkjet or made of a hydrophobic resin forming the pattern.

[0081] According to a second variant, the indication marks may be provided on the frame of the blood component collection and separation medium.

[0082] Optionally or additionally, the frame may have an identification area configured to be able to identify the blood component collection and separation device.

[0083] The identification area may include symbols readable by an automaton or a human.

[0084] According to a specific embodiment, the frame may include at least two lifting elements, preferably four lifting elements, the lifting elements being configured to contact a support on which the blood component collection and separation device is intended to be arranged to lift the blood component collection and separation medium from the support and prevent any contact between the blood component collection and separation medium and the support.

[0085] The present invention further relates to a method for separating and extracting blood components, which implements the blood component collection and separation medium as defined above and implements the following steps:

[0086] · Depositing a whole blood sample in the collection area;

[0087] · Adsorbing the whole blood sample onto a substrate;

[0088] · Retaining red blood cells, white blood cells and platelets on the substrate in the collection area;

[0089] · Diffusing at least one component through the substrate to at least one storage area; and

[0090] · Extracting at least one target component stored in the substrate of the blood component collection and separation medium.

[0091] The method for separating and extracting blood components may further include one or more of the following features.

[0092] According to the first embodiment, the at least one target component is stored in the at least one storage area, and the extraction step of the at least one target component is carried out by perforating at least a part of the at least one storage area or by eluting the at least one target component from the at least one storage area.

[0093] According to the second embodiment, the at least one target component is stored in the collection area, and the extraction step of the at least one target component is carried out by perforating at least a part of the collection area or by eluting the at least one target component from the collection area.

[0094] The whole blood sample deposited in the collection area can have a volume of 50 μL to 500 μL.

[0095] According to a specific embodiment, the target component to be extracted from the whole blood sample can be plasma.

[0096] According to an alternative, the target component to be extracted from the whole blood sample can be DNA, which is stored in white blood cells.

[0097] According to a further alternative, the target component to be extracted from the whole blood sample can be lipids, which are stored in red blood cells.

[0098] According to a specific embodiment, the blood component separation and extraction method can be intended to be implemented by an automaton. Description of the Drawings

[0099] Additional advantages and features will be better identified and understood through the following description and drawings given in an illustrative but non - limiting manner, in which:

[0100] Figure 1 is a schematic top - view of a blood component collection and separation device according to the present invention;

[0101] Figure 2 is in Figure 1 a schematic top - view of a blood component collection and separation medium implemented in the blood component collection and separation device of

[0102] Figure 3A is according to the first specific embodiment of Figure 2 a schematic cross - sectional view of the blood component collection and separation medium of

[0103] Figure 3B is according to the second specific embodiment of Figure 2 a schematic cross - sectional view of the blood component collection and separation medium of

[0104] Figure 4A is according to the first specific embodiment of Figure 1Schematic top view of a blood component collection and separation device;

[0105] Figure 4B is according to a second specific embodiment of Figure 1 Schematic top view of a blood component collection and separation device;

[0106] Figure 5 is according to a specific embodiment of Figure 1 Schematic side view of a blood component collection and separation device;

[0107] Figure 6 is for implementing at least Figure 2 Schematic diagram of a method for separating and extracting blood components of a blood collection and separation medium;

[0108] Figure 7A and 7B is according to the first embodiment of the method, during some steps of Figure 6 Schematic diagram of the top view of a blood component collection and separation device during some steps of a method for separating and extracting blood components;

[0109] Figure 8A and 8B is according to the second embodiment of the method, during some steps of Figure 6 Schematic diagram of the top view of a blood component collection and separation device during some steps of a method for separating and extracting blood components; and

[0110] Figure 9 is after separating blood components on Figure 2 a blood component collection and separation medium, Figures 7A - 8B Schematic diagram of the top view of a blood component collection and separation device. Detailed implementation manners

[0111] In these figures and the following description, the same elements have the same reference numerals.

[0112] In addition, the embodiments in the following description are only considered as examples. Although the description involves one or more embodiments, this does not necessarily mean that each reference numeral relates to the same embodiment, or that the features are only applicable to a single embodiment. The simple features of various embodiments can also be combined to provide new embodiments not explicitly described.

[0113] Hereinafter, it refers to the first and second elements and / or parameters. This indexing is only intended to distinguish two close but different elements and / or parameters. This indexing can be interchanged without affecting the configuration or implementation of those elements and / or parameters. In addition, this indexing is not intended to understand the configuration of those first and second elements and / or parameters in terms of time or space.

[0114] In the following text, the following definitions are used.

[0115] "Whole blood sample" is any blood sample from a human or animal source, stable or unstable, consisting of 55% plasma and 45% formed elements (including erythrocytes (red blood cells), leucocytes (white blood cells), and platelets).

[0116] "Plasma" in whole blood generally includes water and proteins. Typically, plasma contains approximately 92% water, 7% albumin, gamma globulin, antihemophilic factor, and other clotting factors, as well as approximately 1% mineral salts, fats, hormones, and vitamins, with the percentages calculated based on the total weight of the blood.

[0117] "Target component" is a component of whole blood sample 5 that is intended to be titrated after its extraction, and this component is stored in red blood cells 51, platelets 52, white blood cells 53, or plasma 55.

[0118] "Pore size" (in μm) can be determined by American Society for Testing and Materials (ASTM) standard 316 - 03 (2011).

[0119] The maximum pore size, minimum pore size, and mean flow pore size can be measured using a technique called capillary flow porometry. First, the sample of the nonwoven fiber web is wetted with a wetting fluid so that all the pores in the sample are filled. A non - reactive gas with increasing pressure is applied to one side of the wet sample to displace the wetting fluid from the pores. For the wet sample, the gas pressure and gas flow rate downstream of the sample are measured and plotted. After the sample is dried, the test is repeated to plot a similar curve for the dry sample.

[0120] "Maximum pore size" is calculated based on the bubble point (i.e., the gas pressure when air flow through the wet sample is first detected). The term "mean flow pore size" is calculated based on the gas pressure when the flow rate through the wet sample is 50% of the flow rate through the dry sample. The term "minimum pore size" is calculated based on the pressure when the wet flow rate curve merges with the dry flow rate curve. The term "pore size range" is defined as the difference between the "maximum pore size" and the "minimum pore size" (i.e., pore size range = maximum pore size - minimum pore size).

[0121] According to Figure 1 , shows an intention to collect and separate whole blood sample 5 (such as Figures 7A - 8BA blood component collection and separation device 10 for at least one component shown in). The blood component collection and separation device 10 includes at least a blood component collection and separation medium 1 and a frame 11 surrounding the blood component collection and separation medium 1. The frame 11 has a window 19 showing the blood component collection and separation medium 1.

[0122] The blood component collection and separation device 10 has a length M and a width N. For example, according to Figure 1 a specific embodiment, the length M of the blood component collection and separation device 10 can be 75 to 100 mm, and its width N can be 40 to 60 mm. According to one aspect, the length M and width N of the blood component collection and separation device 10 can be configured to be compatible with an automaton, and more specifically, compatible with the dimensions of the rack of such an automaton designed to accommodate the blood component collection and separation device 10 and the fixture designed to pick up the blood component collection and separation device 10, so as to allow, for example, the automation of the blood deposition task. Therefore, the length M and width N of the blood component collection and separation device 10 can be adapted to the requirements of the automaton. More specifically, the length M and width N of the blood component collection and separation device 10 are determined by the frame 11. Therefore, it is easy to adapt the blood component collection and separation device 10 to the requirements of the automaton without changing the dimensions of the blood component collection and separation medium 1.

[0123] The frame 11 can be made of cardboard (also known as "paperboard"). Advantageously, cardboard is a rigid material, usually low-cost and biodegradable. Such a frame 11 made of cardboard can also reduce the production cost of the blood component collection and separation device 10 and reduce the amount of non-biodegradable waste. In addition, even if the blood component collection and collection device 10 is intended to be used by humans or by an automaton, the device must have a minimum stiffness so that at least the device can be easily picked up and handled by humans or an automaton. For example, the frame 11 using cardboard can easily adapt to the length M and width N of the blood component collection and separation device 10 because such a material is easy to cut. The frame 11 can further be made of other rigid materials (such as wood, plastic, metal).

[0124] According to Figure 1 a specific embodiment, the blood component collection and separation device 10 can be square or rectangular. Such a shape is easy to produce and can be produced quickly and cost-effectively by at least reducing waste. In addition, such a shape can be easily picked up by humans or an automaton and can also be recommended for the automation of blood deposition and separation methods.

[0125] Optionally, as Figure 1As shown, the frame 11 may have an identification area 14 configured to identify the blood component collection and separation device 10 and / or the sample deposited on the blood component collection and separation medium 1. The identification area 14 includes symbols that can be read automatically or by a human. For example, such symbols can be QR codes, barcodes, numbers, names, or any other symbols capable of identifying the sample deposited on the blood component separation and collection device 10. According to one variant, the identification area 14 may include an RFID capable of identifying the blood component collection and separation device 10. For example, in the case of collecting multiple samples from the same or different points of multiple patients, such an identification area 14 can avoid errors between samples.

[0126] Figure 2 The blood component collection and separation medium 1 is shown in more detail. The blood component collection and separation medium 1 includes a substrate 3 and a boundary wall 7.

[0127] In the described embodiment, the substrate 3 is intended and preferably capable of being wetted (e.g., completely wetted) by the whole blood sample 5. The substrate 3 has a maximum flow pore size of 8 μm to 35 μm, and such a maximum flow pore size can retain at least red blood cells 51 in the substrate 3. More specifically, the substrate 3 has a maximum flow pore size configured to retain red blood cells 51 and white blood cells 53 on its surface. According to a specific embodiment, the maximum flow pore size of the substrate 3 may be preferably 9.5 μm to 10.5 μm. Additionally or as another variant, the substrate 3 may further have a minimum flow pore size of 0.5 μm to 2 μm, preferably 0.9 μm to 1.3 μm. Then, additionally or as an alternative, the substrate 3 may have an average flow pore size of 2.5 μm to 5 μm, and preferably 3 μm to 3.5 μm. Using such characteristics of the substrate 3 alone or in combination enables the substrate 3 to separate the plasma 55 (as Figure 9 shown) from other components of the whole blood sample 5 (as Figures 7A - 8B shown) (e.g., red blood cells 51 (as Figure 9 shown), platelets 52 (as Figure 9 shown) or white blood cells 53 (also as Figure 9 shown)). In fact, the various components of the whole blood sample 5 have different sizes. For example, the average diameter of red blood cells 51 is 7 μm, the average diameter of platelets 52 is typically 1.5 to 3 μm, the average diameter of white blood cells is 12 μm, and the plasma 55 is mainly composed of water. For example, with such specific pore sizes, red blood cells 51, platelets 52, and white blood cells 53 will be retained on the surface and / or within the substrate 3.

[0128] According to Figure 2In a specific embodiment, the substrate 3 can be a fibrous web comprising a first fiber, a second fiber, and a hydrophilic binder. The first fiber is selected from glass microfibers or synthetic microfibers, and the second fiber is selected from fibrillated fibers. In the art, it is known that glass fibers have a good affinity for red blood cells 53. Therefore, the red blood cells 53 will adhere to the glass fibers, and the movement of the red blood cells is slowed down relative to the movement of the plasma 55 through the substrate 3, which also enables the separation of the components of the whole blood sample 5. However, due to the cutting properties of such glass fibers, some hemolysis of these red blood cells 53 can be observed. To prevent such hemolysis of the red blood cells 53, the hydrophilic binder at least partially covers the first fibers of the fibrous web. It has been shown that 5-7% by weight of the hydrophilic binder in the fibrous web can prevent hemolysis. In addition, the hydrophilic binder can be selected from latex binders, polyvinyl alcohol binders, styrene-butadiene binders, vinyl acetate binders, polysaccharide binders, or protein binders.

[0129] The first fiber can have one or more of the following characteristics: a diameter less than 5 μm, and preferably 0.4 μm to 1 μm, a specific surface area greater than 1.5 m 2 / g, a length / diameter ratio greater than 100, and more preferably greater than 500. Such characteristics of the first fibers of the substrate 3 enable the red blood cells 51 to be intercepted by the substrate 3, and also reduce their diffusion rate through the substrate 3 compared to the plasma 55, and enable the separation of the components forming the whole blood sample 5 through this substrate 3. Such characteristics of the first fibers will have an improved affinity for the red blood cells 51.

[0130] The second fiber can be a cellulose-based fiber, and in particular cotton linter, lyocell fiber, or viscose fiber. According to Figure 2 a specific embodiment, the first fibers and the second fibers forming the substrate 3 can be mixed together. More specifically, the fibrous web can contain 50 to 99% by weight, particularly 70 to 98% by weight, of the first fibers, and 1 to 50% by weight of the second fibers, with 100% by weight corresponding to the fibrous web formed. Such redistribution between the first fibers and the second fibers enables the substrate 3 to be well designed to separate the various components of the whole blood sample 5.

[0131] Optionally, the fibrous web can include at least one additive for improving the characteristics of the substrate 3. For example, the additive can be a stabilizer for biomarkers extracted from the plasma 55, such as animal proteins (such as bovine serum albumin (BSA)) or gelatin derivatives. According to another aspect, the stabilizer can be sucrose to maintain humidity and also preserve the biomarkers.

[0132] In Figures 2 to 3B the embodiment shown, the boundary wall 7 extends at least through the entire depth d of the substrate 3 and forms a pattern 9 in the said substrate 3.Figure 3A and 3B is Figure 2 a cross-sectional view of a blood component collection and separation medium 1, the view being taken along axis C as shown in Figure 2 FIG. According to a specific embodiment shown herein, the depth d of the substrate 3 can be from 50 to 1000 μm, and more preferably from 200 to 500 μm. According to Figure 3A the specific embodiment shown in FIG., the boundary wall 7 only extends through the entire depth d of the substrate 3. In addition, according to Figure 3B the specific embodiment shown in FIG., the boundary wall 7 forming the pattern 9 can extend outside or above one surface of the substrate 3. More specifically, the boundary wall 7 can extend upward from the surface of the substrate 3 to a distance equal to 80% of the depth d of the substrate 3. According to this second specific embodiment, a larger volume of whole blood sample 5 (as shown in Figure 3A FIG.) can be deposited on the substrate 3 compared to the volume that can be provided on the blood component collection and separation medium 1 as shown in Figures 7A - 8B FIG.

[0133] The boundary wall 7 is made of a hydrophobic resin. Using a hydrophobic resin that extends at least through the entire depth d of the substrate 3 enables the whole blood sample 5 to be retained in the pattern 9 and also enables the separation of various components of the whole blood sample 5 in this pattern 9. In addition, using a hydrophobic resin to form the boundary wall 7 enables the plasma 55 to be easily guided without any risk of this component diffusing outside the pattern 9. More specifically, the hydrophobic resin forming the boundary wall 7 can be selected from photocurable resins, more preferably UV-curable resins, or thermosetting resins. According to another embodiment, the hydrophobic resin can be selected from fluorinated resins, modified fluorinated resins, latexes, glycol ether acrylates, acrylates, or combinations thereof. More specifically, the hydrophobic resin can be selected from polymethyl methacrylate resin (PMMA) or polydimethylsiloxane resin (PDMS). For example, such a hydrophobic resin can be applied to the substrate 3 without any contact with the fiber web (e.g., by a non-contact dispensing system). According to another embodiment, for example, the hydrophobic resin can be applied to the substrate 3 by contact with the fiber web (e.g., by screen printing).

[0134] Still referring to Figure 2, the pattern 9 has a collection area 91, at least one storage area 93, and at least one channel 95 connecting the collection area 91 to the at least one storage area 93, and the channel 95 forms a bottleneck between the collection area 91 and the storage area 93. The collection area 91 is intended to receive a whole blood sample 5. In addition, the at least one storage area 93 is intended to store at least one component in the whole blood sample 5 after separating the whole blood sample 5 through the substrate 3. The bottleneck formed by the at least one channel 95 guides the plasma 55 separated from the whole blood sample 5 to the at least one storage area 93, and the separation of the plasma 55 from the whole blood sample 5 is carried out through the substrate 3, which will be explained in more detail below. In addition, the at least one channel 95 enables the whole blood sample 5 to be redistributed throughout the collection area 91 regardless of where the deposition area of the whole blood sample 5 is in the collection area 91. Therefore, the pattern 9 formed by the boundary wall 7 can position the whole blood sample 5 at any position in the collection area 91, and then the diffusion of various components of the whole blood sample 5 is controlled by the substrate 3 (at least due to its porosity) and the at least one channel 95.

[0135] The size of the collection area 91 can be designed to receive a whole blood sample 5 with a volume of 50 to 500 μL. In addition, the collection area 91 can be intended to be perforated to allow the collection of components of the whole blood sample 5 (such as Figures 7A - 8B shown). The collection area 91 of the pattern 9 can have a substantially parallelogram shape, and the at least one storage area 93 can further have a substantially circular shape. In addition, the at least one storage area 93 can be arranged at the corner 97 of the substantially parallelogram shape of the collection area 91. The pattern 9 can have as many storage areas 93 as the collection area 91 has corners 97.

[0136] According to a specific embodiment, the collection area 91 can further contain a salt capable of causing at least partial shrinkage of the red blood cells 51. The shrinkage of the red blood cells 51 enables the improvement of the separation speed of the components of the whole blood sample 5. The salt can be selected from the group consisting of halides and sulfates of alkali metals and alkaline earth metals, and hydrochlorides of organic bases, and more particularly selected from calcium chloride, potassium sulfate, or guanine hydrochloride. The salt can be present in the substrate 3 at a concentration of 1 - 25% of the total weight of the substrate 3, and more particularly 10 - 20% of the total weight of the substrate 3.

[0137] The storage area 93 can be intended to be perforated and / or eluted when the components (e.g., plasma 55) of the whole blood sample 5 intended to be extracted have diffused into the storage area 93. In addition, according to Figure 2 the specific embodiment shown, the at least one storage area 93 can have a pre-perforation 94 facilitating its perforation. Such a pre-perforation can simplify the automation of the perforation process for recovering the plasma that has diffused into the storage area 93.

[0138] According to Figure 2In the illustrated specific embodiment, the pattern 9 has a collection area 91 having a square shape and four storage areas 93 provided at each corner 97 of the collection area 91. The length j of the collection area 91 is 13 mm, the diameter e of the storage area 93 is 8 mm, and such dimensions of the collection area 91 and the storage area 93 are particularly suitable for a 100 μL whole blood sample 5. In addition, each storage area 93 has a circular shape and is connected to the collection area 91 through a channel 95 that forms a bottleneck between the collection area 91 and the associated storage area 93. The at least one channel 95 has a length l that can be from 1 mm to 5 mm and a width w that is from 1 mm to 3 mm. Due to technical limitations, the width w of the channel 95 is implemented to be at least 1 mm. In fact, it is now impossible to obtain a width w of the channel 95 that is less than 1 mm.

[0139] Figure 4A and 4B FIG. illustrates a blood component collection and separation device 10 according to the first and second specific embodiments. According to those specific embodiments, the frame 11 includes means for enabling an automaton (or a human user) to identify the configuration of the blood component collection and separation device 10 in a rack.

[0140] More specifically, according to Figure 4A the first specific embodiment shown, the frame 11 has a triangle 12 printed on one corner, and the triangle 12 is intended to enable an automaton to identify the orientation of the blood component collection and separation device 10. Thus, the triangle 12 can enable the automation of the blood collection and separation method. In fact, the automaton can include the length M and width N of the blood component collection and separation device 10 (as Figure 1 shown) and the position of the collection area 91, and detecting the triangle 12 through a visual device (such as a camera) can enable the automaton to detect the position of the collection area 91.

[0141] In addition, according to the second specific embodiment, as Figure 4B shown, the frame 11 can have a concave corner 13. The concave corner 13 is intended to enable an automaton to identify the orientation of the blood component collection and separation device 10. In fact, for the first specific embodiment, it is known that the concave corner 13 is used to determine and / or identify the position of the blood component collection and separation device 10 (or any other flat object) by an automaton.

[0142] Still referring to Figure 4B the specific embodiment, the blood component collection and separation device 10 further includes at least two indication marks 15 of the collection area 91. These indication marks 15 are configured to enable an automaton to detect the collection area 91, as will be explained in more detail below. According to Figure 4BSpecific implementation embodiments, the indication signs 15 are provided on the blood component collection and separation medium 1. In this case, these indication signs 15 can be printed by inkjet or made of a hydrophobic resin that forms a pattern 9 during the manufacturing process of the blood component collection and separation medium 1. According to a variant not shown, those indication signs 15 can be provided on the frame 11 of the blood component collection and separation device 10. According to this specific implementation embodiment, the indication signs 15 are cross-shaped marks. However, other designs for these indication signs 15 not shown herein can be easily conceived.

[0143] Figure 5 Illustrated is a Figure 1 blood component collection and separation device 10 according to another specific implementation embodiment, wherein the frame 11 can further include at least two lifting elements 16, more specifically four lifting elements 16. The lifting elements 16 are configured to contact a support 17 (such as a table) on which the blood component collection and separation device 10 is intended to be disposed, to lift the blood component collection and separation medium 1 from the support 17 and prevent any contact between the blood component collection and separation medium 1 and the support 17. Thus, these lifting elements 16 can prevent the blood component collection and separation medium 1 from being contaminated by any compounds that may be present on the support 17. For example, such lifting elements 16 can be very useful when the blood component collection and separation device 10 is intended to receive a whole blood sample 5 of more than 200 μL. In fact, the blood component collection and separation medium 1 (as Figure 2 shown) may bend during the deposition of such a volume of whole blood sample 5 thereon, and contact between the support 17 and the blood component collection and separation medium 1 must be prevented. For example, these lifting elements 16 can be made of the same component as the frame 11.

[0144] Now referring to Figures 6 to 9 illustrates a blood component separation and extraction method 100. The method 100 implements the blood component separation and collection medium 1 defined with reference to Figure 2 . According to multiple embodiments, the target component to be extracted from the whole blood sample can be, for example, plasma 55, DNA stored in white blood cells 53, platelets 52, or lipids stored in red blood cells 51.

[0145] The blood component separation and extraction method 100 implements a deposition step S1 of the whole blood sample 5 in the collection area 91. Figure 7A And 8A illustrates this deposition step S1 better. More specifically, this deposition step can be performed at any position in the collection area 91 without any extraction of the blood components of the whole blood sample 5. This deposition can be performed substantially at the center of the collection area 91 (as Figure 7Aas shown), or at another location (such as Figure 8A as shown). This free location for depositing the whole blood sample 5 can simplify the deposition step S1, and also allows reducing the time required to perform such a step and allows potential automation of the deposition step S1. According to Figure 7A and 8A in the specific embodiment shown, the whole blood sample 5 deposited in the collection area 91 has a volume of 50 μL to 500 μL, and more preferably 100 μL.

[0146] Then, the blood component separation and extraction method 100 performs the adsorption step S3 of the whole blood sample 5 onto the substrate 3. During this adsorption step S3, the blood components of the whole blood sample 5 start to be separated. At the same time, the blood component separation and extraction method 100 performs the retention step S5 of red blood cells 51, platelets 52, and white blood cells 53 on the substrate 3 in the collection area 91. In addition, still in parallel with those adsorption S3 and retention S5 steps, the blood component separation and extraction method 100 performs the diffusion S7 of at least one component through the substrate 3 to at least one storage area 93. Figure 7B and 8B illustrate these different steps. More specifically, the arrows F in these figures show the diffusion direction of the whole blood sample 5 in the collection area 91. As described above, the various components of the whole blood sample 5 will not have the same diffusion rate through the substrate 3 of the blood collection and separation medium 1, and this difference allows the separation of the blood components of the whole blood sample 5.

[0147] Figure 9 illustrates the blood component collection and separation device 10 after these adsorption S3, retention S5, and diffusion S7 steps. After these steps, the whole blood sample 5 is separated into various components of the whole blood sample 5. More specifically, red blood cells 51, platelets 52, and white blood cells 53 remain in the collection area 91, and plasma 55 is in the storage area 93. Therefore, it is easy to collect the components of the expected whole blood sample 5. As Figure 9 shown, regardless of where the whole blood sample 5 is deposited (see Figure 7A and 8A), all of the patterns 9 will contain multiple components of the whole blood sample 5. The diffusion of this sample throughout the pattern 9 formed by the boundary walls 7 is due to the channels 95 connecting the storage area 93 to the collection area 91. Additionally, after these steps, the blood component separation and extraction method 100 implements an extraction step S9 of at least one target component stored in the substrate 3 of the blood component collection and separation medium 1. According to a first specific embodiment, the at least one target component may be stored in the at least one storage area 93, and the extraction step S9 of the at least one target component is performed by perforating at least a portion of the at least one storage area 93 or by eluting the at least one target component from the at least one storage area 93. According to a second specific embodiment, the at least one target component may be stored in the collection area 91, and the extraction step S9 of the at least one target component is performed by perforating at least a portion of the collection area 91 or by eluting the at least one target component from the collection area 91. According to Figure 9 a specific embodiment, the target component is plasma 55 and extraction by perforation is intended. To this end, the storage areas 93 have pre-perforations 94 to facilitate the perforation of these areas and thus improve the efficiency of the extraction step S9.

[0148] Furthermore, the blood component separation and extraction method 100 may be intended to be implemented by an automaton. In this case, the deposition step S1 is directly implemented by the automaton, and the extraction step S9 is directly executed by the automaton after a given time, which depends, for example, on the volume of the whole blood sample 5 or the porosity of the substrate 3 forming the blood component collection and separation medium 1. In this case, the blood component collection and separation device 10 has at least two indication marks 15 (as Figure 4B shown) to enable the automaton to detect the collection area 91. More specifically, when there are only two indication marks, the collection area 91 may be arranged between the indication marks 15. The form or characteristics of these indication marks 15 may be imposed by the customer, for example, to adapt to the automaton he sells.

[0149] The above-described multiple embodiments are examples given in an illustrative rather than a restrictive manner. In fact, without departing from the teachings of this specification, those skilled in the art can envision other ranges for the length M and width N of the blood component collection and separation device 10; or adjust the flow aperture of the substrate 3, the depth d of the substrate 3. In addition, those skilled in the art can adjust the types and properties of the additives intended to be added to the fiber web to change the properties of the substrate 3. Then, without departing from the teachings of the embodiments disclosed above, the sizes of the collection area 91, the storage area 93, and the channels 95 can be changed to accommodate the volume of the whole blood sample 5, and whole blood samples 5 of larger volumes can also be envisioned. In addition, without departing from this teaching, the number of the indication signs 15 and their shapes can be changed. Then, other elements other than the triangle 12 or the concave corner 13 can be envisioned to determine the position of the blood component collection and separation device 10.

[0150] Thus, due to the blood component collection and separation medium 1 as disclosed above, the deposition and extraction of blood components in the whole blood sample 5 can be simplified. In fact, the pattern 9 including at least one channel 95 formed by the boundary wall 7 enables the plasma 55 to be guided to at least one storage area 93. In addition, this pattern 9 allows for the simplification of the deposition of the whole blood sample 5 because it only requires setting the whole blood sample 5 anywhere in the collection area 91, then separating the various components through the substrate 3, and allowing these components to diffuse in the multiple areas of the pattern 9 through at least one channel 95. In addition, such a blood component collection and separation medium 1 can automate the blood component separation and extraction method 100.

Claims

1. A blood component collection and separation medium (1), comprising a substrate (3) capable of being wetted by a whole blood sample (5), said substrate (3) having a maximum flow pore size of 8 μm to 35 μm, such a maximum flow pore size enabling at least red blood cells (51) to be retained in said substrate (3), characterized in that The blood component collection and separation medium (1) further includes boundary walls (7) that at least extend through the entire depth (d) of the substrate (3) and form a pattern (9) in the substrate (3), the boundary walls (7) being made of a hydrophobic resin, and wherein the pattern (9) has: · A collection area (91) of the pattern (9) having a predetermined shape, the collection area (91) being intended to receive a whole blood sample (5); · At least one storage area (93), the storage area (93) being intended to collect at least one component in the whole blood sample (5) after separating the whole blood sample (5) through the substrate (3); and · At least one channel (95) connecting the collection area (91) to the storage area (93), the channel (95) forming a bottleneck between the collection area (91) and the storage area (93), wherein the pattern (9) has as many storage areas (93) as the collection area (91) has channels (95); wherein the substrate (3) is a fibrous web comprising a first fiber and a second fiber, the first fiber being selected from glass microfibers or synthetic microfibers, and the second fiber comprising cellulose.

2. The blood component collection and separation medium (1) according to claim 1, wherein the substrate (3) has an average flow pore size of 2.5 μm to 5 μm.

3. The blood component collection and separation medium (1) according to claim 1 or 2, wherein the hydrophobic resin forming the boundary walls (7) is selected from photocurable resins or thermosetting resins.

4. The blood component collection and separation medium (1) according to claim 3, wherein the photocurable resin is selected from UV curable resins.

5. The blood component collection and separation medium (1) according to claim 1 or 2, wherein the hydrophobic resin is selected from fluorinated resins, modified fluorinated resins, latexes, glycol ether acrylates, acrylates, or combinations thereof.

6. The blood component collection and separation medium (1) according to claim 1 or 2, wherein the collection area (91) of the pattern (9) has a parallelogram shape.

7. The blood component collection and separation medium (1) according to claim 6, wherein the storage area (93) is provided at a corner (97) of the parallelogram shape of the collection area (91).

8. The blood component collection and separation medium (1) according to claim 7, wherein the pattern (9) has as many storage areas (93) as the collection area (91) has corners (97).

9. The blood component collection and separation medium (1) according to claim 1 or 2, wherein the storage area (93) has a circular shape.

10. The blood component collection and separation medium (1) according to claim 1 or 2, wherein the channel (95) has a length (l) of 1 mm to 5 mm and a width (w) of 1 mm to 3 mm.

11. The blood component collection and separation medium (1) according to claim 1 or 2, wherein the fibrous web further includes: · A hydrophilic binder.

12. The blood component collection and separation medium (1) according to claim 11, wherein the hydrophilic binder is selected from latex binders.

13. The blood component collection and separation medium (1) according to claim 11, wherein the hydrophilic binder is selected from polyvinyl alcohol binders.

14. The blood component collection and separation medium (1) according to claim 11, wherein the hydrophilic binder is selected from styrene-butadiene binders.

15. The blood component collection and separation medium (1) according to claim 11, wherein the hydrophilic binder is selected from vinyl acetate binders.

16. The blood component collection and separation medium (1) according to claim 11, wherein the hydrophilic binder is selected from polysaccharide binders.

17. The blood component collection and separation medium (1) according to claim 11, wherein the hydrophilic binder is selected from protein binders.

18. The blood component collection and separation medium (1) according to claim 11, wherein the fibrous web comprises 5 - 7% by weight of the hydrophilic binder.

19. The blood component collection and separation medium (1) according to claim 1 or 2, wherein the boundary wall (7) forming the pattern (9) extends beyond one surface of the substrate (3) and extends beyond one surface of the substrate (3) up to a distance equal to 80% of the depth (d) of the substrate (3).

20. The blood component collection and separation medium (1) according to claim 1 or 2, wherein the collection zone (91) further contains a salt capable of causing at least partial shrinkage of red blood cells (51), and the salt is selected from the group consisting of halides and sulfates of alkali metals and alkaline earth metals, and hydrochlorides of organic bases.

21. A blood component collection and separation device (10) for separating and collecting at least one component in a whole blood sample (5), wherein the blood component collection and separation device (10) comprises at least the blood component collection and separation medium (1) according to any one of claims 1 to 20 and a frame (11) surrounding the blood component collection and separation medium (1).

22. A blood component separation and extraction method (100), wherein the blood component separation and extraction method (100) employs the blood component collection and separation medium (1) according to any one of claims 1 to 20, and the blood component separation and extraction method (100) comprises the following steps: · A deposition step (S1) in which the whole blood sample (5) is deposited in the collection zone (91); · An adsorption step (S3) in which the whole blood sample (5) is adsorbed onto the substrate (3); · A retention step (S5) in which red blood cells (51), platelets (52), and white blood cells (53) are retained in the substrate (3) in the collection zone (91); · A diffusion step (S7) in which at least one component diffuses through the substrate (3) to the storage zone (93); and · An extraction step (S9) of extracting at least one target component stored in the substrate (3) of the blood component collection and separation medium (1).

23. The blood component separation and extraction method (100) according to claim 22, wherein the at least one target component is stored in the storage area (93), and the extraction step (S9) of the at least one target component is performed by perforating at least a part of the storage area (93) or by eluting the at least one target component from the storage area (93).

24. The blood component separation and extraction method (100) according to claim 22 or 23, wherein the at least one target component is stored in the collection area (91), and the extraction step (S9) of the at least one target component is performed by perforating at least a part of the collection area (91) or by eluting the at least one target component from the collection area (91).

25. The blood component separation and extraction method (100) according to claim 22 or 23, wherein the whole blood sample (5) deposited in the collection area (91) has a volume of 50 μL to 500 μL.

26. The blood component separation and extraction method (100) according to claim 22 or 23, wherein the target component to be extracted from the whole blood sample (5) is plasma (55).

27. The blood component separation and extraction method (100) according to claim 22 or 23, wherein the target component to be extracted from the whole blood sample (5) is DNA, and the DNA is stored in the white blood cells (53).

28. The blood component separation and extraction method (100) according to claim 22 or 23, wherein the target component to be extracted from the whole blood sample (5) is lipid, and the lipid is stored in the red blood cells (51).

Images