Blood analysis platform

Abstract

The present invention provides a blood analysis platform comprising: a blood separation chip in which whole blood mixed with a magnetic fluid is injected into a separation space formed in the center thereof, and the injected whole blood is separated into blood cells and plasma in the separation space; and a blood separation means which has the blood separation chip received inside the center thereof and provides an external force so as to enable the whole blood received in the blood separation chip to be separated into blood cells and plasma. Thus, without using a syringe injection pump, whole blood not diluted in a buffer may be separated into blood cells and plasma, and the blood may be analyzed using the separated blood cells and plasma.

Description

Blood analysis platform

[0001] The present invention relates to a blood analysis platform, and more specifically, to a blood analysis platform that can easily separate whole blood into blood cells and plasma using a magnetic field and centrifugal force, react the separated plasma with a detector, and analyze the blood based on the degree of aggregation of the blood cells and the degree of color change by the detector.

[0002] Generally, since blood plays a vital role in supplying oxygen, nutrients, hormones, and substances to the cells of all organs in the body, as well as serving as a defense mechanism against infection and transporting toxins and waste products, changes occurring within the body lead to changes in blood composition.

[0003] Therefore, in order to accurately diagnose, treat, and determine the prognosis of a disease, various components of the blood must be separated and tested.

[0004] Technologies for separating plasma from whole blood include centrifugation, filter separation, or technologies using microfluidic chips.

[0005] However, when these separation technologies were applied to plasma separation, the purity of the separated plasma was poor, and excessive physical stimulation was applied to the blood cells, often leading to hemolysis of the blood cells.

[0006] When hemolysis of blood cells occurs, hemoglobin, potassium ions, proteins, nucleic acids, etc. contained in the cells spread into the plasma, which affects the analysis of blood components and leads to erroneous results.

[0007] In the case of microfluidic control technology currently under development, hemolysis occurs less frequently than in centrifugation or filter separation because plasma is separated by inertia and the interaction between blood cells and their microstructures.

[0008] However, it has the disadvantages of poor purity and yield of the separated plasma, and a very slow separation speed.

[0009] In addition, there is also the disadvantage that it is difficult to use whole blood as is, so the blood must be diluted in a buffer before use.

[0010] Therefore, there is a need to develop a fluid separation technology that can separate plasma quickly and with high purity without causing hemolysis of blood cells.

[0011] For prior art, refer to Published Patent No. 10-2021-0010619 (January 27, 2021).

[0012] The present invention aims to provide a blood analysis platform capable of separating blood cells and plasma from whole blood that has not been diluted in a buffer, without using an infusion pump such as a syringe, and analyzing the blood using the separated blood cells and plasma.

[0013] A blood analysis platform according to the present invention comprises: a blood separation chip in which whole blood is injected into a separation space formed in the center and the injected whole blood is separated into blood cells and plasma in the separation space; and a blood separation means in which the blood separation chip is accommodated in the center and which provides an external force to separate the whole blood accommodated in the blood separation chip into blood cells and plasma.

[0014] At this time, the whole blood contained in the separation space of the blood separation chip according to the present invention is mixed with a magnetic fluid.

[0015] And the external force provided by the blood separation means according to the present invention to the blood separation chip is a magnetic field and a centrifugal force.

[0016] Here, the blood separation chip according to the present invention comprises a chip body having a separation space formed in the center for accommodating whole blood, and a separation channel communicating with the separation space and having a length in the outer direction, which allows plasma separated from the separation space to move outward.

[0017] At this time, it is preferable that a plurality of separation channels according to the present invention are arranged radially with respect to the center of the separation space of the chip body.

[0018] In addition, a capillary section having a narrower width than the width of another length can be formed in the portion of the length of the separation channel according to the present invention that is adjacent to the separation space.

[0019] In addition, a ferromagnetic mesh for removing magnetic particles may be provided in the portion of the length of the separation channel according to the present invention that is adjacent to the separation space.

[0020] In addition, it includes a detection unit provided at any point along the length of the separation channel according to the present invention, which reacts with a biomarker contained in plasma to change color.

[0021] The blood separation means according to the present invention comprises a spin body that accommodates the blood separation chip in a receiving space formed in the center, a plurality of magnets that are radially provided on the spin body with respect to the center of the receiving space to form a magnetic field, a cover coupled to the upper side of the spin body to prevent the magnets from detaching, a fixing member provided in the receiving space of the spin body to prevent the blood separation chip accommodated in the receiving space from moving, and a pair of handles provided at the upper and lower centers of the spin body to form an axis so that the spin body can rotate about an axis along a vertical line in the up-and-down direction.

[0022] At this time, a receiving space of the spin body according to the present invention is formed with a seating portion on which the blood separation chip is seated, and it is preferable that the seating portion has a height lower than the middle height of the total height of the receiving space.

[0023] And, according to the present invention, it is preferable that the plurality of magnets are arranged so that the magnetic field forms a Hallbach array.

[0024] In addition, the fixing member according to the present invention may be provided as a bearing.

[0025] The effects exhibited by the blood analysis platform according to the present invention are as follows.

[0026] At a low cost, without using infusion pumps such as syringes, blood cells and plasma can be separated from whole blood that has not been diluted in buffer, and the separated blood cells and plasma can be rapidly analyzed.

[0027] FIG. 1 is an exemplary diagram showing a blood analysis platform according to an embodiment of the present invention.

[0028] FIG. 2 is an exemplary diagram showing a blood separation chip and a blood separation means of a blood analysis platform according to an embodiment of the present invention.

[0029] FIG. 3 is an exploded view showing the disassembled state of a blood analysis platform according to an embodiment of the present invention.

[0030] FIG. 4 is an exemplary diagram showing a state in which magnets are arranged in a Hallbach array according to an embodiment of the present invention.

[0031] FIG. 5 is a graph showing the results of measuring the distance the magnetic solution moved to the center according to the seating height of the blood separation chip accommodated in the receiving space of the blood separation means according to an embodiment of the present invention.

[0032] Figure 6 is an image showing the separation speed results using a blood analysis platform according to an embodiment of the present invention.

[0033] Figure 7 is an image showing the results of the separation of blood cells in whole blood with different hematocrit ratios using a blood analysis platform according to an embodiment of the present invention.

[0034] Figure 8 is an image showing the state in which a biomarker contained in plasma is detected by a detection unit using a blood analysis platform according to an embodiment of the present invention.

[0035] The present invention provides a blood analysis platform comprising a blood separation chip in which whole blood is injected into a separation space formed in the center and the injected whole blood is separated into blood cells and plasma in the separation space, and a blood separation means in which the blood separation chip is accommodated inside the center and an external force is provided to separate the whole blood accommodated in the blood separation chip into blood cells and plasma.

[0036] Hereinafter, preferred embodiments according to the present invention will be described in detail with reference to the attached drawings. Prior to this, terms and words used in this specification and claims should not be interpreted as being limited to their ordinary or dictionary meanings, and should be interpreted in a meaning and concept consistent with the technical spirit of the present invention, based on the principle that the inventor can appropriately define the concept of the terms to best describe his invention.

[0037] Therefore, the embodiments described in this specification and the configurations illustrated in the drawings are merely the most preferred embodiments of the present invention and do not represent all of the technical ideas of the present invention; thus, it should be understood that there may be equivalent variations that can replace them at the time of filing this application.

[0038] The present invention relates to a blood analysis platform in which a blood separation chip containing whole blood containing a magnetic fluid is placed in a blood separation means that forms a magnetic field with a plurality of magnets, and the blood separation means is rotated using a user's finger to separate the whole blood contained in the blood separation chip into blood cells and plasma using the magnetic field and centrifugal force of the blood separation means, and the separated plasma moves along a separation channel and reacts with a detector provided in the separation channel to easily detect and analyze biomarkers (proteins, nucleic acids, etc.) contained in the plasma based on the degree of color change according to the biomarker. Referring to the drawings, the details are as follows.

[0039] Referring to FIGS. 1 to 4, a blood analysis platform according to an embodiment of the present invention includes a blood separation chip (100) and a blood separation means (200) for separating and analyzing blood into plasma and blood cells. First, whole blood mixed with a magnetic fluid is injected into a separation space formed in the center of the blood separation chip (100), and the injected whole blood is separated into blood cells and plasma in the separation space (111).

[0040] The above-mentioned blood separation chip (100) can be examined in more detail as follows.

[0041] A blood separation chip (100) according to an embodiment of the present invention includes a chip body (110) that accommodates whole blood and a separation channel (120) through which plasma separated from blood cells in whole blood moves.

[0042] At this time, the chip body (110) is made of a transparent material through which the interior is visible, has a polygonal shape overall, has a certain height to accommodate whole blood inside, and a separation space (111) is formed in the center (=center) of the chip body (110) to accommodate whole blood injected from the outside.

[0043] The above separation space (111) communicates with the blood injection hole (112) formed on the upper surface of the chip body (110) so that whole blood can be injected from the outside.

[0044] Therefore, whole blood can be injected into the separation space (111) inside the center of the chip body (110) through the blood injection port (112).

[0045] Here, the above whole blood is mixed with a magnetic fluid, so that the blood cells contained in the blood acquire paramagnetic properties, and separation from the plasma is achieved by external forces (magnetic field and centrifugal force).

[0046] In addition, the separation channel (120) is formed to have a length extending outward relative to the center of the separation space (111) located in the center of the chip body (110), so that the plasma separated from the blood cells moves outward along the length of the separation channel (120).

[0047] Here, the portion of the length of the separation channel (120) adjacent to the separation space (111) forms a capillary section (121) having a narrower width than the other length width of the separation channel (120), thereby facilitating the separation of liquid plasma from whole blood.

[0048] In addition, a ferromagnetic mesh (122) is provided in the portion of the length of the separation channel (120) adjacent to the separation space (111) so that magnetic particles contained in the magnetic fluid are filtered.

[0049] At this time, the ferromagnetic mesh (122) may be made of a ferromagnetic material such as iron, nickel, or cobalt, but preferably it is made of nickel that does not rust.

[0050] And the separation channel (120) can be formed in at least two or more multiple units, and the multiple separation channels (120) are arranged in a radial shape with respect to the center of the separation space (111) and communicate with the separation space (111).

[0051] The number of the above separation channels (120) can be formed in correspondence with the number of biomarkers to be analyzed.

[0052] And the inner side of the length of the separation channel (120) is connected to the separation space (111), and a discharge hole (124) is formed on the outer side of the length of the separation channel (120).

[0053] Therefore, plasma that has moved to the separation channel (120) through the discharge port (124) can be easily discharged to the outside.

[0054] In addition, at any point along the length of the separation channel (120), a detection unit (123) is provided that reacts with a biomarker (protein, nucleic acid, etc.) contained in the plasma moving along the separation channel (120).

[0055] Here, when a plurality of separation channels (120) are formed in the blood separation chip (100), each separation channel (120) may be equipped with a detection unit (123) that reacts with different factors, thereby allowing for the detection of various substances at once.

[0056] And the blood separation means (200) according to an embodiment of the present invention accommodates the blood separation chip (100) inside and provides external forces, such as a magnetic field and centrifugal force, to separate the whole blood accommodated in the blood separation chip (100) into blood cells and plasma.

[0057] Looking more closely at the blood separation means (200), the blood separation means (200) includes a spin body (210), a magnet (220), and a cover (230). The spin body (210) has an overall cylindrical shape and forms a receiving space (211) in the center for receiving the blood separation chip (100).

[0058] At this time, the upper side of the receiving space (211) is formed as an opening to allow the blood separation chip (100) to enter and exit, and the lower side of the receiving space (211) is formed as a seating portion (212) where the blood separation chip (100) is seated and fixed.

[0059] Here, the mounting portion (212) is rotatably installed, and when the mounting portion rotates, the blood separation chip (100) also rotates accordingly.

[0060] This embodiment is configured such that the blood separation chip (100) rotates according to the rotation of the mounting portion (212), but a turntable (not shown) may be provided so that the spin body (210) can rotate around the center of the blood separation chip (100), and the spin body (210) and the blood separation chip (100) may rotate together with the center of the blood separation chip (100) housed in the receiving space (211) as the axis.

[0061] Additionally, referring to FIG. 5, the mounting portion (212) is positioned at a certain height relative to the bottom surface so that the whole blood contained in the blood separation chip (100) can be maximally affected by the magnetic field.

[0062] It is preferable that the blood separation chip (100) be positioned at a height of 2 to 4 mm above the bottom surface (0 mm) so as to be formed to have a height lower than the middle height at the total height of the above-mentioned receiving space (211).

[0063] More preferably, the separation space (111) and separation channel (120) of the blood separation chip (100) are positioned at a height of 3 mm above the bottom surface (0 mm), so that they have a height.

[0064] In addition, a magnet receiving portion (213) for receiving a magnet (220) is formed around the receiving space (211) of the spin body (210), and it is preferable that the magnet receiving portion (213) be formed in a plurality of radially arranged relative to the receiving space (211).

[0065] At this time, the magnet receiving portions (213) are formed as spaces with a shape corresponding to the outer shape of the magnet (220) to accommodate the magnet (220) forming a cuboid, and the upper side is formed as an opening so that the magnet (220) can enter and exit.

[0066] According to an embodiment of the present invention, the blood separation means (200) is equipped with a total of 8 magnets (220), and the magnet receiving portions (213) in which each magnet (220) is received are formed by being arranged radially with respect to the receiving space (211).

[0067] Accordingly, the eight magnets (220) each housed in the magnet receiving portions (213) form a magnetic field in the spin body (210), so that the blood cells, which are diamagnetic in the whole blood housed in the blood separation chip (100), are pushed by the magnetic field and gather in the center of the separation space (111) of the blood separation chip (100), and the plasma moves outward along the length of the separation channel (120) communicating with the separation space (111), so that the whole blood housed in the separation space (111) of the blood separation chip (100) is separated into blood cells and plasma by the magnetic field of the magnets (220) housed in the magnet receiving portions (213).

[0068] The magnet (220) is made of an electromagnet and can turn off the magnetic field when whole blood is supplied to the blood separation chip (100), and turn on the magnetic field after whole blood is supplied.

[0069] Referring to FIG. 4, it is preferable that the magnets (220) be arranged in a Hallbach array in which the N pole and S pole are orthogonally arranged so as to amplify the magnetic field on one side (upward direction of the spin body (210)) and form the magnetic field on the other side (downward direction of the spin body (210) to be close to 0.

[0070] And a cover (230) is provided on the upper part of the spin body (210) to prevent the magnets (220) each housed in the magnet receiving portions (213) from escaping to the outside.

[0071] The blood separation means (200) according to an embodiment of the present invention may provide centrifugal force along with a magnetic field for blood separation, and provides centrifugal force to the blood separation chip (100) received in the receiving space (211) of the spin body (210) by rotating the spin body (210).

[0072] Referring to FIG. 3, the blood separation means (200) further includes a fixing member (240) and a handle (250) so that the spin body (210) can rotate.

[0073] First, the fixed member (240) is received in the receiving space (211) of the spin body (210) in which the blood separation chip (100) is received, and when the spin body (210) rotates, it controls the flow of the blood separation chip (100) received in the receiving space (211).

[0074] Here, the fixed member (240) may employ a bearing so that the spin body (210) can rotate easily.

[0075] And the handle (250) is a pair and is respectively connected to the upper and lower sides of the mounting portion (212) and the blood separation chip (100), so that the blood separation chip (100) can rotate around an axis along a vertical line in the up-and-down direction.

[0076] At this time, the handles (250) are formed in a conical shape such that the portion facing the blood separation chip (100) is wide and the cross-sectional area becomes smaller as it goes toward the opposite portion (upward or downward).

[0077] The handles (250), respectively attached to the upper and lower centers of the blood separation chip (100), allow the blood separation chip (100) to rotate about an axis along a vertical line in the up-and-down direction. When a user grasps and rotates the handle (250), the blood separation chip (100) and the seating portion (212) rotate together, and centrifugal force is applied to the blood separation chip (100).

[0078] When centrifugal force is applied to the blood separation chip (100), blood cells, which are diamagnetic in the whole blood contained in the separation space (111) of the blood separation chip (100), remain in the separation space (111) due to the magnetic field and centripetal force according to the magnetic field, and plasma moves outward along the radially arranged separation channels (120) due to centrifugal force.

[0079] Additionally, when the fixed member (240) is provided as a bearing, it is provided in the receiving space (211) of the spin body (210) so that the spin body (210) can freely rotate about the center of the receiving space (211) as an axis.

[0080] At this time, the fixed member (240) may use a ball bearing, wherein the ball bearing uses a ball as a rolling element between the outer diameter member and the inner diameter member, and the outer diameter member and the inner diameter member of the fixed member (240) can rotate individually relative to each other by means of the ball as a rolling element.

[0081] Accordingly, when the fixing member (240) is received in the receiving space (211) of the spin body (210), the outer diameter member of the fixing member (240) is fixed to the inner surface of the receiving space (211) of the spin body (210), and the inner diameter member of the fixing member (240) is fixed to the seating portion (212), so that the seating portion (212) and the blood separation chip (100) are installed to rotate with respect to the spin body (210).

[0082] A blood analysis platform according to an embodiment of the present invention comprises a blood separation chip and a blood separation means. When a blood separation chip containing whole blood mixed with a magnetic fluid in the center is placed in a receiving space formed in the center of the blood separation means, blood cells are pushed toward the center and plasma is moved toward the outside by a magnetic field formed by magnets provided on the outer circumference of the blood separation chip, thereby separating the whole blood into blood cells and plasma.

[0083] At this time, the magnets are arranged in a Hallbach array, and the magnets rotate around the center of the blood separation chip as an axis so that the blood cells can be well gathered to the center.

[0084] When the blood separation chip (100) rotates and the plasma moves outward along the separation channel by centrifugal force, a detection unit is provided at the outer edge to measure biomarkers (proteins, nucleic acids, etc.) in the plasma. By providing detection units that detect different biomarkers in each of the multiple separation channels arranged radially, various biomarkers can be detected at once.

[0085] FIG. 5 is a graph showing the results of measuring the distance the magnetic solution moved to the center according to the seating height of the blood separation chip accommodated in the receiving space of the blood separation means according to an embodiment of the present invention. When the height of the total receiving space is 11 mm, the blood separation reaction speed when the blood separation chip is positioned according to the height of the seating part supporting the blood separation chip relative to the bottom surface of the spin body was measured (separation speed measurement over 5 minutes). The distance of movement was measured to be longer at 2 to 4 mm, which is lower than the middle height of the total height of the receiving space, but the distance of movement was longest at 3 mm.

[0086] Therefore, the height of the above-mentioned seating portion is most ideally 3 mm from the bottom surface of the spin body, which is lower than the middle height of the total height of the receiving space.

[0087] Figure 6 shows the results of diamagnetic separation speeds using a blood analysis platform according to an embodiment of the present invention, where A) is a graph showing the results of measuring the separation speed of beads (6, 8.9 μm) of different sizes and cells (MDA-MB-231) according to the concentration of the magnetic solution (SPION), B) is a separation image of a 6 μm bead, C) is a separation image of an 8.9 μm bead, and D) is a separation image of a cell (MDA-MB-231) (Scale bar: 200 μm).

[0088] By examining A) the separation speed graph and B, C, and D) the separated images, it can be confirmed that when substances (diamagnetic) in a magnetic solution are exposed to a magnetic field, the larger their size and density, the faster they move away from the magnet.

[0089] Figure 7 shows the results of the separation of blood cells in whole blood with different hematocrit ratios using a blood analysis platform according to an embodiment of the present invention, where A) is a graph showing the measured values ​​of the blood cell separation area according to a hematocrit (HCT) of 30% to 60%, and B) is a microscopic image (Scale bar: 2mm) 10 minutes after the start of separation at a hematocrit of 30% to 60%.

[0090] Looking at this, it is possible to confirm that the blood cells have gathered towards the center within 10 minutes, regardless of the blood cell volume (normal 40~50%, anemia and hypocytosis with less than 40%, polycytosis with more than 50%: erythrocytosis), and through this, anemia can be analyzed based on the distribution of the gathered blood cells.

[0091] FIG. 8 shows the state of detecting biomarkers (proteins, nucleic acids, etc.) contained in plasma using a detection unit with a blood analysis platform according to an embodiment of the present invention, where A) is a fluorescence image of a control portion that is not treated on the bottom surface of the separation channel, and B) to D) can be seen as fluorescences attached to Alexa Fluor 488-Streptavidin collected after fixing NHS-Biotin (10 mM) on the bottom surface of the separation channel.

[0092] E) is a graph showing a comparison of the fluorescence image intensity (degree of color change) between the control and detection sections.

[0093] Therefore, depending on the detection method located on the outer side of the separation channel, various analysis methods such as ELISA and colorimetric assay can be applied.

[0094] The present invention has been described with reference to embodiments illustrated in the drawings, but this is merely illustrative, and those skilled in the art will understand that various modifications and equivalent alternative embodiments are possible therefrom. Accordingly, the true technical scope of protection of the present invention should be determined by the technical spirit of the appended claims.

Claims

1. A blood separation chip in which whole blood is injected into a separation space formed in the center, and the injected whole blood is separated into blood cells and plasma in the separation space; and A blood analysis platform comprising: a blood separation means that provides an external force to separate whole blood contained in the blood separation chip into blood cells and plasma, wherein the blood separation chip is accommodated in the center.

2. In Claim 1, The whole blood accommodated in the separation space of the above-mentioned blood separation chip is, A blood analysis platform characterized by a mixture of magnetic fluids.

3. In Claim 2, A blood analysis platform characterized in that the external force provided by the above-mentioned blood separation means to the blood separation chip is either a magnetic field or a centrifugal force.

4. In Claim 3, The above blood separation chip is, A chip body having a separation space formed in the center for accommodating whole blood, and A blood analysis platform comprising a separation channel that communicates with the separation space and has an external length, allowing plasma separated in the separation space to move outward.

5. In Claim 4, The above separation channel is, A blood analysis platform characterized by multiple units arranged radially in length with respect to the center of the separation space of the chip body.

6. In Claim 4, A blood analysis platform characterized by forming a capillary section having a narrower width than the width of another length in the portion of the length of the separation channel adjacent to the separation space.

7. In Claim 4, In the portion of the length of the above separation channel adjacent to the above separation space, A blood analysis platform characterized by having a ferromagnetic mesh that removes magnetic particles from plasma moving along the length of the separation channel.

8. In Claim 4, Provided at any point along the length of the above separation channel, A blood analysis platform comprising a detector that changes color by reacting with a biomarker contained in plasma.

9. In Claim 3, The above blood separation means is A spin body that accommodates the above-mentioned blood separation chip in a receiving space formed in the center, and A plurality of magnets radially provided on the spin body with respect to the center of the above-mentioned receiving space to form a magnetic field, and A cover coupled to the upper side of the spin body to prevent the magnet from detaching, and A fixing member provided in the receiving space of the spin body above to prevent flow of the blood separation chip received in the receiving space, and A blood analysis platform comprising a pair of handles provided at the upper and lower centers of the blood separation chip, forming an axis so that the blood separation chip can rotate along an vertical axis in the up-and-down direction.

10. In Claim 9, In the receiving space of the spin body above, A mounting portion is formed on which the above blood separation chip is placed, and A blood analysis platform characterized by the above-mentioned seating portion having a height lower than the intermediate height at the total height of the above-mentioned receiving space.

11. In Claim 9, The above plurality of magnets, A blood analysis platform characterized by magnetic fields arranged to form a Hallbach array.

12. In Claim 9, The above fixing member is A blood analysis platform characterized by being equipped with a bearing.

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