Blood component collection and separation medium, blood component collection and separation device including the medium, and blood component separation and extraction method using the medium

Abstract

The present invention relates to a blood component collection and separation medium (1), comprising a substrate (3) having a maximum flow pore size, the maximum flow pore size enabling 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 area (91); at least one storage area (93), the storage area (93) being intended to collect at least one component in a 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 present invention further relates to a blood component collection and separation device and a blood component separation and extraction method.

Description

Technical Field

[0001] 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, the use of the medium in blood separation and extraction methods, 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 used as a blood component collector and separator. Background Art

[0002] Biological samples are often used in laboratory and clinical settings to analyze various components in samples (e.g., blood samples). Biological samples are often processed in liquid form. Therefore, liquid samples are collected, processed in collection devices, transported to laboratories, and stored. There are various problems with activities around liquid samples (e.g., blood), including the risk of container breakage or leakage (which leads to sample loss and potential contamination and / or infection risks), sample instability during transportation and storage, and transportation vehicle limitations associated with the transportation of liquid biohazardous materials.

[0003] In order to avoid such problems, various devices and methods for drying samples have been developed. Blood as a dry sample is not considered a biohazard material and can be transported by conventional delivery services like any other package. To this end, a blood sample (usually a few drops) is placed on paper and allowed to dry. In addition, in order to improve the efficiency of these devices and methods, some papers are configured to separate blood components.

[0004] In fact, blood consists of two parts: blood cells and the fluid in which the blood cells are suspended. This fluid is called blood plasma ("plasma"), and it is a proteinaceous fluid that can 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 - The main eukaryotic cell types present in blood are red blood cells (erythrocytes), white blood cells (leucocytes), and platelets, although other cell types may also be detected. Red blood cells make up about half of the volume of a blood sample.

[0005] For rapid diagnosis of plasma components in clinical, hospital and field conditions, it is very important to separate red blood cells from plasma in blood samples, because red blood cells can impair the sensitivity of the analysis. Whole blood can also cause undesirable chemical interferences. For example, hemoglobin released from red blood cells can affect the performance of certain clinical analyses due to the iron heme group (which can act as a catalyst).

[0006] In the process of separating the red blood cells from the fluid fraction, it is more important that the red blood cells do not lyse or rupture, which could result in the release of internal components into the plasma and also contaminate the plasma sample. To obtain plasma from whole blood for testing, essentially four different types of techniques are used: namely those based on gravity, pressure drop, capillary flow and centrifugal force.

[0007] The assay should be cheap to perform under field conditions and the samples should be disposable. Some of the above mentioned techniques do not meet these goals very well. Pressure drop and centrifugal force require specialized equipment, and the implementation of pumps and centrifuges makes them unsuitable for field use. Gravity-based techniques are also too slow for field use.

[0008] Therefore, assays have been developed for separation of whole blood into red blood cells and plasma, which are based on capillary flow in lateral flow assay devices.Many such assays are disclosed in the art.

[0009] U.S. Patent Nos. 4,477,575 and 4,816,224 describe media that use a layer of glass microfibers to separate red blood cells from whole blood. Paper containing 100% glass microfibers of the type proposed is inherently fragile and requires great care in handling. Although strength can be increased by using adhesives, some adhesives can interfere with analysis or render the sheet hydrophobic.

[0010] Another approach involves using a single layer of media made from a composite material of glass microfibers, cellulose fibers and synthetic staple fibers. In this regard, reference is made to U.S. Patent No. 5,186,843.

[0011] Although blood separation media proposed in the past are suitable for their intended purposes, some improvements are still needed.

[0012] Typically, filtration on lateral flow assay devices is hampered by insufficient speed because whole blood is quite viscous and begins to coagulate in air. The pores of standard blood filters 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 efficiency of the filtration. This also reduces the number of pores available for plasma to pass through and reduces the total amount of plasma that passes through the filter per unit time.

[0013] Furthermore, damage or rupture of the cells may occur. This is particularly true if the fibers constituting the nonwoven are glass, such as the glass proposed in US Pat. No. 5,186,843.

[0014] Due to the affinity of red blood cells to 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 capillaries. However, even if red blood cells show good affinity to glass fibers, those glass fibers may also increase hemolysis, and the separation of multiple blood components may not be so effective. In order to solve the problem of this prior art, a blood collection and separation medium that prevents this hemolysis is known from the international patent application WO2017 / 017314 submitted by the applicant. This hemolysis is also prevented by using a resin intended to at least partially coat the glass fibers to prevent hemolysis. However, even if this blood collection and separation medium provides good separation results of multiple blood components, the deposition of whole blood samples must also be completed accurately, and the area to be, for example, perforated or eluted 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 for an automated method for collecting and separating blood components from a whole blood sample. Summary of the invention

[0015] The present invention is directed to at least partially addressing the shortcomings of the prior art discussed above by providing a blood component collection and separation medium that facilitates deposition of whole blood samples.

[0016] Another aspect of the present invention is to provide a blood component collection and separation medium, which 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 prevents hemolysis during the process of separating the multiple components in the whole blood sample.

[0017] Another aspect of the present invention which is different from those described above is to provide a blood component collection and separation device which can be used in an automated method.

[0018] In order to at least partially meet at least one of the objectives set forth above, the present invention relates to a blood component collection and separation medium, the blood component collection and separation medium comprising a substrate intended to be wetted by a whole blood sample, the substrate having a maximum flow pore size of 8 μm to 35 μm, such maximum flow pore size enabling at least red blood cells to be trapped in the substrate, the blood component collection and separation medium further comprising a boundary wall patterned in the substrate, the boundary wall being made of a hydrophobic resin, and the pattern having:

[0019] A collection area, which is intended to receive a whole blood sample;

[0020] At least one storage area intended to separate at least one component of a whole blood sample by means of a substrate

[0021] storing at least one component of the whole blood sample; and

[0022] - At least one channel connecting the collection zone to at least one storage zone, said channel forming a bottleneck between the collection zone and the storage zone.

[0023] The presence of a channel forming a bottleneck between the collection area and the storage area enables the plasma to diffuse to at least one storage area and trap the red blood cells in the collection area. Therefore, the identification of the area containing the specific components of the whole blood sample can be easily identified. In addition, the deposition of the whole blood sample can be carried out anywhere in the collection area without affecting the diffusion of the plasma in the storage area, which also simplifies the deposition process of the whole blood sample on the blood component collection and separation medium.

[0024] The blood component collection and separation medium according to the present invention may further have one or more of the following features alone or in combination.

[0025] According to a specific embodiment, the maximum flow pore size of the substrate is preferably 9.5 μm to 10.5 μm.

[0026] 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.

[0027] According to a specific embodiment, the substrate may have a mean flow pore size of 2.5 μm to 5 μm.

[0028] According to this specific embodiment, the mean flow pore size is preferably 3 μm to 3.5 μm.

[0029] The hydrophobic resin forming the boundary wall may be selected from a photocurable resin, more preferably a UV curable resin, or a thermosetting resin.

[0030] More specifically, the hydrophobic resin may be selected from fluorinated resins, modified fluorinated resins, latex, glycol ether acrylates, acrylates, or combinations thereof.

[0031] According to a specific embodiment, the hydrophobic resin is selected from polymethyl methacrylate (PMMA) or polydimethylsiloxane (PDMS).

[0032] According to a specific embodiment, the collection area of ​​the pattern can have a substantially parallelogram shape.

[0033] According to this particular embodiment, the at least one storage area may be arranged at a corner of the substantially parallelogram-shaped collecting area.

[0034] According to a variant of this particular embodiment, the pattern may have as many storage areas as the collection areas have corners.

[0035] Alternatively or additionally, the at least one storage area may have a substantially circular shape.

[0036] According to a first embodiment, said storage area is intended to be perforated.

[0037] According to a second embodiment, at least one component stored in the storage area is intended to be eluted.

[0038] According to a third embodiment, said collecting zone is intended to be perforated.

[0039] 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.

[0040] According to a specific embodiment, the substrate may be a fiber web comprising:

[0041] a first fiber selected from glass microfibers or synthetic microfibers;

[0042] a second fiber selected from fibrillated fibers; and

[0043] ·Hydrophilic adhesive.

[0044] The hydrophilic binder may be selected from latex binders.

[0045] According to one alternative, the hydrophilic binder may be chosen from polyvinyl alcohol binders.

[0046] According to a further alternative, the hydrophilic binder may be chosen from styrene-butadiene binders.

[0047] According to another alternative, the hydrophilic binder may be chosen from vinyl acetate binders.

[0048] According to one variant, the hydrophilic binder may be chosen from polysaccharide binders.

[0049] According to another variant, the hydrophilic binder may be chosen from protein binders.

[0050] According to a specific embodiment, the first fibers may have a diameter of less than 5 μm, and preferably from 0.4 μm to 1 μm.

[0051] Alternatively or in addition, the first fiber may have a length greater than 1.5 m 2 / g specific surface area.

[0052] Additionally, the first fibers may have a length / diameter ratio greater than 100, and more preferably greater than 500.

[0053] The first fibers and the second fibers may be mixed together.

[0054] According to a particular embodiment, the second fibers are cellulose-based fibers, and in particular cotton linters, lyocell fibers or viscose fibers.

[0055] According to the specific embodiments described above, the fiber web may include 5 to 7 weight percent of a hydrophilic binder.

[0056] According to one specific embodiment, the fiber web may comprise 50 to 99% by weight, in particular 70 to 98% by weight, of first fibers, and 1 to 50% by weight of second fibers, 100% by weight corresponding to the formed fiber web.

[0057] The hydrophilic binder may at least partially cover the first fibers of the fiber web.

[0058] According to one aspect, the fiber web may include at least one additive for improving the properties of the substrate.

[0059] According to one embodiment of this aspect, the at least one additive may be a stabilizer for the biomarker extracted from plasma, such as an animal protein (such as bovine serum albumin (BSA)), a gelatin derivative or sucrose.

[0060] According to a specific embodiment, some boundary walls or all 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.

[0061] According to one embodiment, the boundary walls forming the pattern may extend beyond one surface of the substrate.

[0062] The collection zone may be sized to receive a whole blood sample volume of 50 to 500 μL.

[0063] As an alternative or in addition, the collection zone may further contain a salt capable of achieving at least partial shrinkage of the erythrocytes, the salt being selected from the group of halides and sulfates of alkali metals and alkaline earth metals, and hydrochlorides of organic bases.

[0064] According to this alternative, the salt may be chosen from calcium chloride, potassium sulfate or guanine hydrochloride.

[0065] Other suitable salts are manganese chloride, potassium chloride, magnesium chloride and sodium chloride.

[0066] The salt may be present in the substrate in a concentration of 1-25% of the total weight of the substrate, more particularly 10-20% of the total weight of the substrate.

[0067] According to one particular embodiment, the pattern may have a collection zone having a square shape and four storage zones arranged at each corner of the collection zone, each storage zone having a circular shape and connected to the collection zone by a channel forming a bottleneck between the collection zone and the associated storage zone.

[0068] The at least one storage area may have pre-perforations to facilitate perforation thereof.

[0069] The present invention further relates to a blood component collecting and separation device intended to collect and separate at least one component in a whole blood sample, the blood component collecting and separation device comprising a blood component collecting and separation medium at least as defined above and a frame surrounding the blood component collecting and separation medium.

[0070] The blood component collection and separation device according to the present invention may further include one or more of the following features.

[0071] According to a particular embodiment, the frame may be made of cardboard.

[0072] The frame may enable the blood component collection and separation device to be picked up and handled by a robot.

[0073] The blood component collection and separation device may be square or rectangular.

[0074] According to a first specific embodiment, the frame may have a triangle printed on one corner, the triangle being intended to enable the robot to recognize the orientation of the blood component collection and separation device.

[0075] According to a second specific embodiment, the frame may have a notched angle intended to enable the robot to recognize the orientation of the blood component collection and separation device.

[0076] Additionally, the blood component collection and separation device may have a length and width configured to be compatible with a robot.

[0077] Optionally, the blood component collection and separation device may have at least two indicator marks of the collection area, the indicator marks being configured to enable an automated machine to detect the collection area.

[0078] According to a first variant, the indicator marks may be provided on the blood component collection and separation medium outside the pattern.

[0079] According to this first variant, the indicator marks may be printed by inkjet or made of a patterned hydrophobic resin.

[0080] According to a second variant, the indicator mark may be provided on the frame of the blood component collection and separation medium.

[0081] Alternatively or additionally, the frame may have an identification region configured to enable identification of the blood component collection and separation device.

[0082] The identification area may include a symbol that is readable by an automated machine or a human.

[0083] According to a specific embodiment, the frame may include at least two lifting elements, preferably four lifting elements, which are configured to contact the support on which the blood component collection and separation device is intended to be arranged, so as 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.

[0084] The present invention further relates to a method for separating and extracting blood components, the method implementing the blood component collection and separation medium as defined above, and implementing the following steps:

[0085] Deposit the whole blood sample in the collection area;

[0086] Adsorb the whole blood sample onto the substrate;

[0087] · Retaining red blood cells, white blood cells and platelets on the substrate in the collection zone;

[0088] diffusing at least one component through the substrate to at least one reservoir; and

[0089] Extracting at least one target component stored in the matrix of the blood component collection and separation medium.

[0090] The blood component separation and extraction method may further include one or more of the following features.

[0091] According to a 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 performed by perforating at least a portion of the at least one storage area or by eluting the at least one target component from the at least one storage area.

[0092] According to a second embodiment, the at least one target component is stored in a collection area and the extraction step of the at least one target component is performed by perforating at least a portion of the collection area or by eluting the at least one target component from the collection area.

[0093] The whole blood sample deposited in the collection area may have a volume of 50 μL to 500 μL.

[0094] According to a specific embodiment, the target component to be extracted from the whole blood sample may be plasma.

[0095] According to one alternative, the target component to be extracted from the whole blood sample may be DNA, which is stored in white blood cells.

[0096] According to a further alternative, the target component intended to be extracted from the whole blood sample may be a lipid, which is stored in erythrocytes.

[0097] According to a particular embodiment, the blood component separation and extraction method may be intended to be performed by an automatic machine. BRIEF DESCRIPTION OF THE DRAWINGS

[0098] Additional advantages and features will be better recognized and understood from the following description given by way of illustration and not limitation, and the accompanying drawings, in which:

[0099] Figure 1 is a schematic diagram of a top view of a blood component collection and separation device according to the present invention;

[0100] Figure 2 is Figure 1 A schematic diagram of a top view of a blood component collection and separation medium implemented in a blood component collection and separation device;

[0101] Figure 3A According to the first specific embodiment Figure 2 A schematic cross-sectional view of a blood component collection and separation medium;

[0102] Figure 3B According to the second specific embodiment Figure 2 A schematic cross-sectional view of a blood component collection and separation medium;

[0103] Figure 4A According to the first specific embodiment Figure 1 A schematic top view of a blood component collection and separation device;

[0104] Figure 4B According to the second specific embodiment Figure 1 A schematic top view of a blood component collection and separation device;

[0105] Figure 5 According to a specific implementation plan Figure 1 A schematic side view of a blood component collection and separation device;

[0106] Figure 6 Is to implement at least Figure 2 Schematic diagram of a method for separating and extracting blood components using blood collection and separation media;

[0107] Fig. 7A and 7B According to a first embodiment of the method, Figure 6 A schematic diagram of a top view of a blood component collection and separation device during some steps of a blood component separation and extraction method;

[0108] Fig. 8A and 8B According to a second embodiment of the method, Figure 6 A schematic diagram of a top view of a blood component collection and separation device during some steps of a blood component separation and extraction method; and

[0109] Fig. 9 is Figure 2 After the blood components are collected and separated on the separation medium, Figures 7A-8B Schematic diagram of a top view of a blood component collection and separation device. DETAILED DESCRIPTION

[0110] In the drawings and the following description, like elements have like reference numerals.

[0111] In addition, the embodiments in the following description are considered to be examples only. Although the description relates to one or more embodiments, this does not necessarily mean that each figure mark relates to the same embodiment, or that a feature applies only to a single embodiment. Simple features of various embodiments may also be combined to provide new embodiments that are not explicitly described.

[0112] In the following, it refers to the first and second elements and / or parameters. This indexation is intended only to distinguish between two close but not identical elements and / or parameters. This indexation can be interchanged without affecting the configuration or implementation of those elements and / or parameters. In addition, this indexation is not intended to understand the configuration of those first and second elements and / or parameters in time or space.

[0113] In the following, the following definitions are used.

[0114] A "whole blood sample" is any blood sample, from human or animal origin, stable or unstable, consisting of 55% plasma and 45% formed elements, including erythrocytes (red blood cells), leucocytes (white blood cells), and platelets.

[0115] The "plasma" in whole blood usually includes water and protein. Typically, plasma contains about 92% water, 7% albumin, gamma globulin, antihemophilic factor and other coagulation factors, and about 1% mineral salts, fats, hormones and vitamins, the percentages being calculated based on the total weight of the blood.

[0116] The “target component” is a component of the whole blood sample 5 that is intended to be titrated after its extraction, and this component is stored in the red blood cells 51 , the platelets 52 , the white blood cells 53 , or the plasma 55 .

[0117] “Pore size” (in μm) can be determined by American Society for Testing and Materials (ASTM) standard 316-03 (2011).

[0118] The maximum pore size, minimum pore size and mean flow pore size can be measured using a technique called capillary flow porometry. First, a sample of the nonwoven web sample is wetted with a wetting fluid so that all the holes in the sample are filled. A non-reactive gas with increased pressure is applied to one side of the wet sample to displace the wetting fluid from the holes. For the wet sample, the gas pressure and gas flow rate downstream of the sample are measured and plotted. After the sample is dry, the test is repeated to plot a similar curve for the dry sample.

[0119] The "maximum pore size" is calculated based on the bubble point (i.e., the gas pressure at which air flow through the wet sample is first detected). The term "mean flow pore size" is calculated based on the gas pressure at which the flow through the wet sample is 50% of the flow through the dry sample. The term "minimum pore size" is calculated based on the pressure at which the wet flow rate curve is merged 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).

[0120] according to Figure 1 , showing a method for collecting and separating a whole blood sample 5 (such as Figures 7A-8B The blood component collecting and separating device 10 is a device for collecting and separating at least one component of the blood component (shown). The blood component collecting and separating device 10 includes at least a blood component collecting and separating medium 1 and a frame 11 surrounding the blood component collecting and separating medium 1. The frame 11 has a window 19 showing the blood component collecting and separating medium 1.

[0121] The blood component collection and separation device 10 has a length M and a width N. For example, according to Figure 1According to a specific embodiment, the length M of the blood component collection and separation device 10 can be 75 to 100 mm, and the width N thereof 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 automatic machine, and more specifically, compatible with the dimensions of a shelf of such an automatic machine intended to accommodate the blood component collection and separation device 10 and a clamp intended to pick up the blood component collection and separation device 10, so as to allow automation of tasks such as blood deposition. Therefore, the length M and width N of the blood component collection and separation device 10 can be adapted to the requirements of the automatic machine. 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 automatic machine without changing the size of the blood component collection and separation medium 1.

[0122] The frame 11 can be made of cardboard (also called "paperboard"). Advantageously, cardboard is a rigid material that is generally 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 separation device 10 is intended to be used by humans or by an automatic machine, the device must have a minimum rigidity that at least enables the device to be easily picked up and handled by humans or automatic machines. For example, the use of cardboard for the frame 11 can easily adapt to the length M and width N of the blood component collection and separation device 10 because such material is easy to cut. The frame 11 can be further composed of other rigid materials (e.g., wood, plastic, metal).

[0123] according to Figure 1 In 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 further be produced quickly and in a cost-effective manner by at least reducing waste. In addition, such a shape can be easily picked up by humans or automatic machines and can also be recommended for automation of blood deposition and separation methods.

[0124] Optionally, as Figure 1As shown, the frame 11 can have an identification area 14, which is configured to be able 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 a symbol that can be read by an automatic machine or a human. For example, such a symbol can be a QR code, a bar code, a number, a name, or any other symbol that can identify the sample deposited on the blood component collection and separation device 10. According to a variant, the identification area 14 can include an RFID that can identify the blood component collection and separation device 10. For example, in the case of collecting multiple samples from the same point or different points of multiple patients, such an identification area 14 can make it possible to avoid errors between samples.

[0125] Figure 2 The blood component collection and separation medium 1 is shown in more detail. The blood component collection and separation medium 1 comprises a base 3 and a boundary wall 7.

[0126] In the described embodiment, the substrate 3 is intended to and preferably is 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, such a maximum flow pore size is capable of retaining 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. In addition or as an additional variation, 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, in addition 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. Such features of the substrate 3, used alone or in combination, enable the substrate 3 to retain plasma 55 (such as Fig. 9 ) and other components of whole blood sample 5 (such as Figures 7A-8B As shown) (e.g., red blood cells 51 (e.g. Fig. 9 As shown), platelets 52 (as Fig. 9 ) or WBC53 (also as Fig. 9 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 generally 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 a specific pore size, red blood cells 51, platelets 52 and white blood cells 53 will be trapped on the surface and / or in the substrate 3.

[0127] according to Figure 2In a specific embodiment, the substrate 3 can be a fiber mesh including a first fiber, a second fiber and a hydrophilic adhesive. The first fiber is selected from glass microfibers or synthetic microfibers, and the second fiber is selected from fibrillated fibers. It is known in the art that glass fibers have a good affinity for red blood cells 51. Therefore, the red blood cells 51 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 characteristics of such glass fibers, some hemolysis of these red blood cells 51 can be observed. In order to prevent this hemolysis of the red blood cells 51, the hydrophilic adhesive at least partially covers the first fiber of the fiber mesh. It has been shown that 5-7% by weight of the hydrophilic adhesive in the fiber mesh can prevent hemolysis. In addition, the hydrophilic adhesive can be selected from a latex adhesive, a polyvinyl alcohol adhesive, a styrene-butadiene adhesive, a vinyl acetate adhesive, a polysaccharide adhesive or a protein adhesive.

[0128] The first fiber may 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, and 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 erythrocytes 51 to be trapped by the substrate 3, and also reduce their diffusion rate through the substrate 3 compared to the plasma 55, and enable the components forming the whole blood sample 5 to be separated by the substrate 3. Such characteristics of the first fibers will have an improved affinity with the erythrocytes 51.

[0129] The second fiber may be a cellulose-based fiber, and in particular cotton linters, lyocell fibers or viscose fibers. Figure 2 In a specific embodiment, the first fibers and the second fibers forming the substrate 3 can be mixed together. More specifically, the fiber mesh can contain 50 to 99% by weight, especially 70 to 98% by weight of the first fibers, and 1 to 50% by weight of the second fibers, 100% by weight corresponding to the formed fiber mesh. This redistribution between the first fibers and the second fibers enables the substrate 3 to be well designed to separate multiple components of the whole blood sample 5.

[0130] Optionally, the fiber web may include at least one additive for improving the characteristics of the substrate 3. For example, the additive may be a stabilizer for the biomarkers extracted from the plasma 55, such as an animal protein such as bovine serum albumin (BSA) or a gelatin derivative. According to another aspect, the stabilizer may be sucrose to maintain humidity and also preserve the biomarkers.

[0131] exist Figures 2 to 3B In 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 said substrate 3 . Figure 3A and 3B yes Figure 2 A cross-sectional view of a blood component collection and separation medium 1, the view being based on Figure 2 According to the specific embodiment shown here, the depth d of the substrate 3 can be 50 to 1000 μm, and more preferably 200 to 500 μm. Figure 3A In the particular embodiment shown, the boundary wall 7 extends only through the entire depth d of the base 3. In addition, according to Figure 3B In the embodiment shown, the boundary wall 7 forming the pattern 9 may extend outside or above one surface of the substrate 3. More specifically, the boundary wall 7 may extend upward from the surface of the substrate 3 to a distance equal to 80% of the depth d of the substrate 3. Figure 3A Compared with the volume set on the blood component collection and separation medium 1 shown, a larger volume of whole blood sample 5 can be deposited on the substrate 3 (such as Figures 7A-8B shown).

[0132] The boundary wall 7 is made of a hydrophobic resin. The use of a hydrophobic resin extending at least through the entire depth d of the substrate 3 enables the whole blood sample 5 to be kept in the pattern 9, and also enables the separation of multiple components of the whole blood sample 5 in the pattern 9. In addition, the use of a hydrophobic resin to form the boundary wall 7 enables the plasma 55 to be easily guided without any risk of the component diffusing outside the pattern 9. More specifically, the hydrophobic resin forming the boundary wall 7 can be selected from a photocurable resin, more preferably a UV curable resin, or a thermosetting resin. According to another embodiment, the hydrophobic resin can be selected from a fluorinated resin, a modified fluorinated resin, a latex, a glycol ether acrylate, an acrylate, or a combination thereof. More specifically, the hydrophobic resin can be selected from a polymethyl methacrylate resin (PMMA) or a polydimethylsiloxane resin (PDMS). For example, such a hydrophobic resin can be applied to the substrate 3 without any contact with the fiber mesh (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 contacting the fiber mesh (e.g., by screen printing).

[0133] Still refer 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, the channel 95 forming a bottleneck between the collection area 91 and the storage area 93. The collection area 91 is intended to receive the whole blood sample 5. In addition, the at least one storage area 93 is intended to store at least one component of the whole blood sample 5 after the whole blood sample 5 is separated by 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 by the substrate 3, which will be explained in more detail below. In addition, the at least one channel 95 makes it possible to redistribute the whole blood sample 5 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 makes it possible to set the whole blood sample 5 at any position in the collection area 91, and then the diffusion of the 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.

[0134] The size of the collection area 91 can be designed to receive a volume of 50 to 500 μL of a whole blood sample 5. In addition, the collection area 91 can be designed to be perforated to allow the collection of components of the whole blood sample 5 (such as Figures 7A-8B ). The collection area 91 of the pattern 9 may have a substantially parallelogram shape, and the at least one storage area 93 may further have a substantially circular shape. In addition, the at least one storage area 93 may be arranged at a corner 97 of the substantially parallelogram shape of the collection area 91. The pattern 9 may have as many storage areas 93 as the collection area 91 has corners 97.

[0135] According to a specific embodiment, the collection area 91 may further contain a salt capable of achieving at least partial shrinkage of the red blood cells 51. The shrinkage of the red blood cells 51 makes it possible to improve the separation rate of the components of the whole blood sample 5. The salt may be selected from the group 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 may 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.

[0136] The storage area 93 may be perforated and / or eluted when the components of the whole blood sample 5 (e.g., plasma 55) to be extracted have diffused into the storage area 93. Figure 2 In the specific embodiment shown, the at least one storage area 93 may have a pre-perforated hole 94 for facilitating its perforation. Such pre-perforation can simplify the automation of the perforation process for recovering plasma that has diffused into the storage area 93.

[0137] according to Figure 2In the specific embodiment shown, the pattern 9 has a collection area 91 having a square shape and four storage areas 93 arranged at each corner 97 of the collection area 91. The length j of the collection area 91 is 13 mm, and the diameter e of the storage area 93 is 8 mm. Such dimensions of the collection area 91 and the storage area 93 are particularly suitable for 100 μL of whole blood sample 5. In addition, each storage area 93 has a circular shape and is connected to the collection area 91 by 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 1 mm to 5 mm, and a width w of 1 mm to 3 mm. Due to technical limitations, the width w of the channel 95 is implemented as at least 1 mm. In fact, it is currently impossible to obtain a width w of the channel 95 that is less than 1 mm.

[0138] Figure 4A and 4B Blood component collection and separation devices 10 according to first and second embodiments are illustrated. According to those embodiments, the frame 11 includes means enabling, for example, an automated machine (or a human user) to recognize the configuration of the blood component collection and separation device 10 in a rack.

[0139] More specifically, according to Figure 4A In the first embodiment shown, the frame 11 has a triangle 12 printed on one corner, which is intended to enable the robot to recognize the orientation of the blood component collection and separation device 10. Thus, the triangle 12 enables the automation of the blood collection and separation method. In practice, the robot may include a length M and a width N (e.g. Figure 1 As shown) and the position of the collection area 91, and detecting the triangle 12 by a visual device (such as a camera) enables the automatic machine to detect the position of the collection area 91.

[0140] Furthermore, according to a second specific embodiment, Figure 4B As shown, the frame 11 may have a concave corner 13. The concave corner 13 is intended to enable the robot to identify the direction 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 the robot.

[0141] Still refer to Figure 4B In a specific embodiment of the blood component collection and separation device 10, at least two indicator marks 15 of the collection area 91 are further included. These indicator marks 15 are configured to enable the robot to detect the collection area 91, as explained in more detail below. Figure 4BIn a specific embodiment, the indicator marks 15 are arranged on the blood component collection and separation medium 1. In this case, these indicator marks 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 indicator marks 15 can be arranged on the frame 11 of the blood component collection and separation device 10. According to this specific embodiment, the indicator marks 15 are cross-shaped marks. However, other designs for these indicator marks 15 not shown herein can be easily conceived.

[0142] Figure 5 According to another specific embodiment, Figure 1 A blood component collection and separation device 10, wherein the frame 11 may further include at least two lifting elements 16, more specifically four lifting elements 16. The lifting elements 16 are configured to be in contact with a support 17 (e.g., a table) on which the blood component collection and separation device 10 is intended to be arranged, so as 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. Therefore, these lifting elements 16 are able to prevent any contamination of the blood component collection and separation medium 1 by compounds that may be present on the support 17. For example, such lifting elements 16 can be very useful in the case where the blood component collection and separation device 10 is intended to receive more than 200 μL of whole blood sample 5. In fact, the blood component collection and separation medium 1 (e.g., Figure 2 The support 17 (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 composition as the frame 11.

[0143] Reference now Figures 6 to 9 , illustrating a blood component separation and extraction method 100. The method 100 implements the reference Figure 2 The defined blood component collection and separation medium 1. According to various embodiments, the target component to be extracted from the whole blood sample may be, for example, plasma 55, DNA stored in white blood cells 53, platelets 52, or lipids stored in red blood cells 51.

[0144] The blood component separation and extraction method 100 implements a deposition step S1 of the whole blood sample 5 in the collection area 91 . Fig. 7A and 8A More specifically, the deposition step S1 can be performed at any location in the collection area 91 without any extraction of blood components of the whole blood sample 5. The deposition can be performed substantially in the center of the collection area 91 (e.g. Fig. 7A), or at another location (as shown Fig. 8A ). Such a free position for depositing the whole blood sample 5 can simplify the depositing step S1 and also allow reducing the time required to perform such a step and allowing potential automation of the depositing step S1. Fig. 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.

[0145] Then, the blood component separation and extraction method 100 implements the adsorption step S3 of the whole blood sample 5 onto the substrate 3. The blood components of the whole blood sample 5 begin to be separated during this adsorption step S3. At the same time, the blood component separation and extraction method 100 implements the interception step S5 of the 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 steps S3 and interception steps S5, the blood component separation and extraction method 100 implements the diffusion step S7 of at least one component through the substrate 3 to at least one storage area 93. Figure 7B and 8B These different steps are illustrated. More specifically, arrows F in these figures show the diffusion direction of the whole blood sample 5 in the collection area 91. As previously mentioned, the various components of the whole blood sample 5 will not have the same diffusion speed through the substrate 3 of the blood collection and separation medium 1, and this difference allows the blood components of the whole blood sample 5 to be separated.

[0146] Fig. 9 The blood component collection and separation device 10 is shown after these adsorption step S3, interception step S5, and diffusion step S7. After these steps, the whole blood sample 5 is separated into the various components of the whole blood sample 5. More specifically, the red blood cells 51, platelets 52, and white blood cells 53 remain in the collection area 91, and the plasma 55 is in the storage area 93. Therefore, it is easy to collect the desired components of the whole blood sample 5. Fig. 9 As shown, no matter where the whole blood sample 5 is deposited (see Fig. 7A and 8A), all patterns 9 will contain multiple components of the whole blood sample 5. The diffusion of the sample throughout the pattern 9 formed by the boundary wall 7 is due to the channel 95 connecting the storage area 93 to the collection area 91. In addition, 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 can 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 can 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 Fig. 9 In the specific embodiment of the invention, the target component is plasma 55 and is intended to be extracted by perforation. To this end, the storage area 93 has pre-perforations 94 to facilitate the perforation of these areas and thereby improve the efficiency of the extraction step S9.

[0147] Furthermore, the blood component separation and extraction method 100 may be intended to be implemented by an automatic machine. In this case, the deposition step S1 is directly implemented by the automatic machine, and the extraction step S9 is directly performed by the automatic machine after a given time, which depends on, for example, 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 indicator marks 15 (such as Figure 4B ), so that the automatic machine can detect the collection area 91. More specifically, when there are only two indicator marks, the collection area 91 can be arranged between the indicator marks 15. The form or characteristics of these indicator marks 15 can be applied by the customer, for example, in order to adapt it to the automatic machine he sells.

[0148] The multiple embodiments described above are examples given in an illustrative manner rather than a restrictive manner. In fact, without departing from the teachings of this specification, those skilled in the art may conceive of other ranges of 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 may adjust the type and properties of the additives intended to be added to the fiber mesh 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 channel 95 may be changed to accommodate the volume of the whole blood sample 5, and a larger volume of whole blood sample 5 may also be conceived. In addition, without departing from the present teachings, the number of indicator marks 15 and their shapes may be changed. Then, it is conceivable that other elements besides the triangle 12 or the concave angle 13 determine the position of the blood component collection and separation device 10.

[0149] Therefore, 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 the deposition of the whole blood sample 5 to be simplified, because it is only necessary to set the whole blood sample 5 anywhere in the collection area 91, then separate the multiple components by the substrate 3, and allow these components to diffuse in multiple areas of the pattern 9 by at least one channel 95. In addition, this blood component collection and separation medium 1 enables the automation of 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), the 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 trapped in the substrate (3), characterized in that The blood component collection and separation medium (1) further comprises a boundary wall (7) extending at least through the entire depth (d) of the substrate (3) and forming a pattern (9) in the substrate (3), wherein the boundary wall (7) is made of a hydrophobic resin, and wherein the pattern (9) has: A collection area (91) of a pattern (9) of predetermined shape, the collection area (91) being intended to receive a whole blood sample (5); ● more than one storage area (93), the storage area (93) being intended to collect at least one component of the whole blood sample (5) after separation of the whole blood sample (5) by the substrate (3); and more than 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); The substrate (3) is a fiber web comprising first fibers, the first fibers being selected from glass microfibers or synthetic microfibers.

2. The blood component collection and separation medium (1) according to claim 1, wherein the substrate (3) has a mean 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 wall (7) is selected from a photocurable resin or a thermosetting resin.

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

5. The blood component collection and separation medium (1) according to claim 1 or 2, wherein the hydrophobic resin is selected from fluorinated resin, modified fluorinated resin, latex, glycol ether acrylate, acrylate or a combination 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 arranged 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 areas (91) have 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 fiber mesh further comprises: ● second fibers, said second fibers being selected from fibrillated fibers; and ●Hydrophilic adhesive.

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 adhesive is selected from styrene-butadiene adhesives.

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 fiber web comprises 5-7% by weight of a hydrophilic binder.

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

20. A blood component collection and separation medium (1) according to claim 1 or 2, wherein the collection area (91) further contains a salt capable of achieving 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) intended to separate and collect at least one component in a whole blood sample (5), wherein the blood component collection and separation device (10) comprises at least a 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) implements the blood component collection and separation medium (1) according to any one of claims 1 to 20 and wherein the blood component separation and extraction method (100) implements the following steps: ● A deposition step (S1) of depositing the whole blood sample (5) in the collection area (91); ● an adsorption step (S3) of the whole blood sample (5) into the substrate (3); A step (S5) of trapping red blood cells (51), platelets (52) and white blood cells (53) in the substrate (3) in the collection area (91); A diffusion step (S7) of diffusing at least one component through the substrate (3) into the storage area (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. A 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 wherein the extraction step (S9) of the at least one target component is performed by perforating at least a portion of the storage area (93) or by eluting the at least one target component from the storage area (93).

24. A blood component separation and extraction method (100) according to claim 22 or 23, wherein the at least one target component is stored in the collecting area (91), and the extraction step (S9) of the at least one target component is performed by perforating at least a portion of the collecting area (91) or by eluting the at least one target component from the collecting 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 50µL to 500µL.

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, which is stored in the red blood cells (51).

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