KR101750936B1 - Device and Method for Blood Plasma Separation - Google Patents

Abstract

(i) a body portion in which an enclosed space through which blood can flow is formed integrally and a channel portion and a ridge are alternately and continuously formed on an outer surface of the body portion; (ii) an inflow portion through which the blood located above the body portion flows; (iii) a blood-collecting part located at one side of the body; And (iv) a plasma discharging portion located on the other side of the body, wherein the ridge portion is discontinuously formed, a plasma chip for plasma separation comprising the plasma separating chip, the plasma separation And a plasma array for plasma separation, and a plasma separation method using the same.

Description

BACKGROUND OF THE INVENTION 1. Field of the Invention The present invention relates to a plasma separation apparatus,

The present invention relates to a technique for separating blood plasma from blood, and more particularly, to a technique for separating blood plasma from blood by (i) forming a sealed space through which blood can flow, ; (ii) an inflow portion through which the blood located above the body portion flows; (iii) a blood-collecting part located at one side of the body; And (iv) a plasma discharging portion located on the other side of the body, wherein the ridge portion is discontinuously formed, a plasma chip for plasma separation comprising the plasma separating chip, the plasma separation And a plasma array for plasma separation, and a plasma separation method using the same.

The blood is largely divided into a solid component and a liquid component. The solid component includes blood cells such as a red blood cell and a white blood cell, and the liquid component includes a blood plasma. Plasma is defined herein as a liquid component comprising blood platelets, including about 90% by volume of water, about 7 to 8% by volume of plasma protein, lipids, sugars, inorganic salts and non- And includes urea, amino acid, uric acid and the like. The plasma protein mainly includes albumin and globulin, and also contains fibrinogen related to blood coagulation. The lipid may include cholesterol, lecithin, and the like. The inorganic salts may include sodium, chlorine, potassium, calcium, magnesium, etc., and play an important role in maintaining normal osmotic pressure in the body. Since each component and content of plasma can be changed according to the disease state, it can be usefully used for diagnosis of disease or condition of disease. Plasma may also be useful for transfusion of components of blood that are transfused into a patient in need of a useful component contained in the plasma.

Because of this availability, techniques have been developed for separating plasma from the blood. For example, centrifugation, capillary action, and application of electrical pulses in the microfluidic chip at right angles to the direction of blood flow are examples of methods for separating blood cells from blood from blood. However, these conventional plasma separation techniques have problems in that space required for plasma separation is large, space utilization is low, blood and plasma flow are slow in plasma separation process, and plasma separation efficiency is low, and separation accuracy is low, Or more than the reference value.

Therefore, there is a need to develop a technique capable of separating blood plasma from blood with a simple and quick high separation efficiency and accuracy.

Korean Patent Publication No. 10-2010-0117404

Techniques for separating blood plasma from blood with a high separation efficiency and / or accuracy in a simpler and faster manner are proposed herein.

(I) a body portion having a space in which blood can flow therein integrally and having a channel portion and a ridge alternately and continuously formed on an outer surface thereof;

(ii) an inflow portion through which the blood located above the body portion flows;

(iii) a blood-collecting part located at one side of the body part; And

(iv) a plasma outlet part located on the other side of the body part

And a plasma separation chip.

Wherein the ridge portion and the channel portion communicate with each other in the closed space and the ridge portion is inclined at an angle of more than 0 DEG and less than 90 DEG with one side surface of the body while forming a protruding space communicating with the channel portion, The ridge may be discontinuously formed in an oblique direction.

Another example is

(1) a blood injector into which blood is injected;

(2) a first plasma separator connected to the blood injector;

(3) a second plasma separator connected to the first plasma separator; And

(4)

Wherein the first plasma separation part comprises at least one plasma separation chip comprising a continuous ridge and the second plasma separation part comprises at least one plasma separation chip comprising a discontinuous ridge.

Thereby providing a chip array for plasma separation.

Another example provides a plasma separation device comprising the plasma separation chip or the plasma separation chip array.

The plasma separation device may include two or more of the plasma separation chip or the plasma separation chip array.

The plasma separation chip, the plasma array chip and / or the plasma separation device can separate the plasma from the high concentration blood sample diluted 20 times or less by volume of the whole blood or the whole blood with high efficiency and / or accuracy.

Another example provides a plasma separation method using the plasma separation chip, the plasma separation chip array, or the plasma separation apparatus.

In this specification, a plasma separation chip capable of separating plasma from blood with a high separation efficiency and / or accuracy in a simpler and faster manner, a plasma array chip for plasma separation comprising the plasma separation chip, a plasma separation chip and / Or a plasma array for plasma separation, and a plasma separation method using the plasma separation chip, the plasma separation chip array, and / or the plasma separation device.

In one example,

(i) a body portion in which a space through which blood can flow is integrally formed, and a channel portion and a ridge are alternately and continuously formed on an outer surface of the body portion;

(ii) an inflow portion through which the blood located above the body portion flows;

(iii) a blood-collecting part located at one side of the body part; And

(iv) a plasma outlet part located on the other side of the body part

The present invention also provides a plasma-separating chip.

Exemplary forms of the chip for plasma separation provided in this specification are shown in Figs. 11 and 12. Fig. This will be described below with reference to Figs. 11 and 12. Fig.

The space through which the blood can flow in the body part 101 may be formed as a separate space or an upper surface of the body part of the inflow part 301 (the arrow indicates the inflow direction of the main blood) And may be an enclosed space at a portion excluding the portion 401 and the plasma discharging portion 501 and may be integrally formed so that the blood introduced from the inflow portion flows to the blood cell discharging portion and the plasma discharging portion.

The protruding portion 102 (continuous shape; 105: discontinuous shape) refers to a region where a protruding space is formed in a body portion through which blood flows, the channel portion 103 refers to a region between a protruding portion and a protruding portion, The ridge portion and the at least one channel portion are alternately and continuously formed in the flow direction of the blood.

The ridge portion and the channel portion communicate with each other in a space where blood of the body portion can flow. That is, the blood can flow through the ridge portion and the channel portion which are alternately and continuously formed.

The raised portion forms a protruding space protruding outside the body portion. The raised portion has an inner space having a height (depth: h) of the height of the channel portion (depth: h _c ) and the height of the protruding space (depth: h _r ).

The raised portion is formed to be inclined at an angle (?) Of more than 0 DEG and less than 90 DEG (an angle formed by one side surface and ridge portion in the blood movement direction) to one side surface (400) of the body portion.

Wherein the inclined angle of the ridge portion is greater than 0 deg., Less than 90 deg., Less than 90 deg., Less than 90 deg., Less than 20 deg. (Refer to Fig. Less than 90 °, more than 30 ° and less than 90 °, more than 40 ° and less than 90 °, more than 45 ° and less than 90 °, more than 50 ° and less than 90 °, more than 55 ° and less than 90 °, more than 60 ° and less than 90 °, more than 65 ° 90 degrees or less, 75 degrees or more and 90 degrees or less, 80 degrees or more and 90 degrees or less, 85 or more and 90 or less, 0 or more and 85 or less, 10 to 85 or 20 to 85 , 30 to 85, 40 to 85, 45 to 85, 50 to 85, 55 to 85, 60 to 85, 65 to 85, 70 to 85, 75 80 ° to 85 °, more than 0 ° and less than 80 °, 10 ° to 80 °, 20 ° to 80 °, 30 ° to 80 °, 40 ° to 80 °, 45 ° to 80 °, 50 ° 80 °, 55 ° to 80 °, 60 ° to 80 °, 65 ° to 80 °, 70 ° to 80 °, 75 ° to 80 °, more than 0 ° and 75 ° Of the total thickness of the substrate is in the range of 10 to 75 degrees, 20 to 75 degrees, 30 to 75 degrees, 40 to 75 degrees, 45 to 75 degrees, 50 to 75 degrees, 55 to 75 degrees, 60 to 75 degrees, 65 to 75, 70 to 75, 0 to 70, 10 to 70, 20 to 70, 30 to 70, 40 to 70, 45 to 70, 50 60 属 to 70 属, 65 属 to 70 属, more than 0 属 and less than 65 属, 10 属 to 65 属, 20 属 to 65 属, 30 属 to 65 属, 40 属60 属 to 65 属, 45 属 to 65 属, 50 属 to 65 属, 55 属 to 65 属, 60 属 to 65 属, more than 0 属 and less than 60 属, 10 属 to 60 属, 20 属 to 60 属, 60 属, 40 属 to 60 属, 45 属 to 60 属, 50 属 to 60 属, 55 属 to 60 属, more than 0 属 and less than 55 属, 10 属 to 55 属, 20 属 to 55 属, 40 ° to 55 °, 45 ° to 55 °, 50 ° to 55 °, more than 0 ° and less than 50 °, 10 ° to 50 °, 20 ° to 50 °, 30 ° to 50 °, 40 ° to 50 ° , Or 45 [deg.] To 50 [deg.].

The obliquely formed ridge may be discontinuously formed (105) so that the inner space is cut off in an oblique direction to form one or more discontinuous spaces. The disconnection of the internal space of the ridge may include disposing one or more structures (104) such as columns or partitions in the inclined ridge so that the internal space is disconnected in an inclined direction, or the ridge may be discontinuous May be patterned to form more than one space. Each ridge may include two or more disconnected spaces or one or more posts or a barrier structure (wherein two or more spaced spaces may be formed).

One or more ridges may be formed and one or more ridges may not be parallel to each other if they satisfy the angle condition with the one side of the body as described above and may be parallel to each other or between 0 and 30 , 0 ° to 25 °, 0 ° to 20 °, 0 ° to 15 °, 0 ° to 10 °, 0 ° to 5 °, or 0 ° to 3 ° .

The two or more separated spaces formed in the discontinuous ridge may be parallel to each other if they satisfy the angular condition with the one side of the body as described above. In order to increase the efficiency of plasma separation, 0 ° to 5 °, or 0 ° to 3 °, but it is not limited thereto, and may be an angle of 30 ° to 30 °, 0 ° to 25 °, 0 ° to 20 °, 0 ° to 15 °, 0 ° to 10 °, It is not.

As described above, in the discontinuous ridge portion formed to be inclined at a height (depth) higher than the channel portion (deep) (a height (depth) as high as the height of the protruding space), the flow direction and speed of blood are different from that of the channel portion, (Size and size), and the movement of the blood cells is changed, and the blood collects in one direction.

In order to cause the specific hemocyte movement in the ridge to separate the hemocyte component from the plasma component in the blood, the ridge space of the ridge should be at least enough to accommodate all or part of the erythrocytes. Further, when the raised space becomes a space enough for the leukocyte to be sufficiently contained, the red blood cells are not separated and are washed away together with the plasma. Therefore, the height of the raised space (depth: h _r : elevated height obtained by subtracting the channel portion height hc from the inner space of the raised portion) and the width (w _r ) can be appropriately adjusted according to the size of the white blood cell and the red blood cell. In addition, the height (depth) of two or more raised spaces formed in one ridge may be equal to or different from each other. In one example, the height hr of the raised space may be equal to or greater than the height hc of the channel portion. Considering that the average diameter of erythrocytes is about 8-10 micrometers, the thickness is about 2-3 micrometers, and the average diameter of leukocytes is about 15 micrometers, the height (depth: h _r 1 to 16 um, 1 to 15 um, 1 to 14 um, 1 to 13 um, 1 to 12 um, 1 to 11 um, 1 to 18 um, 1 to 18 um, 1 to 17 um, 2 um to 1 um, 2 um to 20 um, 2 um to 19 um, 2 um to 18 um, 2 um to 17 um, 2 um to 16 um, um, 3 to 16 um, 3 to 15 um, 3 to 14 um, 3 to 13 um, 3 to 16 um, 3 to 16 um, um, 4 to 15 um, 4 to 15 um, 4 to 14 um, 4 to 18 um, 4 to 17 um, 4 to 17 um, 4 to 15 um, 4 to 15 um, 3 to 11 um, 3 to 10 um, 4 to 20 um, 4 to 13 um, 4 to 12 um, 4 to 11 um, 4 to 10 um, 5 to 20 um, 5 to 19 um, 5 to 18 um, 5 to 17 5 to 11 um, 5 to 10 um, 6 to 20 um, 6 to 19 um, 6 to 18 um, 5 to 15 um, 5 to 14 um, 5 to 13 um, 5 to 12 um, um, 6 to 17 um, 6 to 16 um, 6 to 15 um, 6 to 14 um, 6 to 13 um, 6 to 12 um, 6 to 11 um, 6 to 10 um, 7 to 20, 7 to 18 um, 7 to 17 um, 7 to 16 um, 7 to 15 um, 7 to 14 um, 7 to 13 um, 7 to 12 um, 7 to 11 um, 7 to 10 um, 8 to 20 um , 8 to 19 um, 8 to 18 um, 8 to 17 um, 8 to 16 um, 8 to 15 um, 8 to 14 um, 8 to 13 um, 8 to 12 um, 8 to 11 um, 8 to 10 um , 9 to 20 um, 9 to 19 um, 9 to 18 um, 9 to 17 um, 9 to 16 um, 9 to 15 um, 9 to 14 um, 9 to 13 um, 9 to 12 um, , Or 9 to 10 [mu] m.

The width w _r of the raised space may be equal to or different from the height of the raised space h _r . In one example, the width w _r may be greater than or equal to the height h _r , Lt; / RTI > Also, the widths of two or more raised spaces formed in one ridge may be the same or different. When two or more raised portions are formed, the widths of the raised portions may be the same or different from each other. For example, the raised area or the raised parts of the width, the height of the raised area of 0.5 to 3 times the (D h _r), from 0.5 to 2.5 times, 0.5 to 2 times, 0.5 to 1.5, 0.7 to 3 times, 0.7 to 2.5 Fold, 0.7 to 2 times, 0.7 to 1.5 times, 1 to 3 times, 1 to 2.5 times, 1 to 2 times, or 1 to 1.5 times, but is not limited thereto.

The height (h _c ) of the channel portion may be such that a blood cell component such as red blood cells (which is larger than red blood cells in the case of white blood cells but is amorphous and can be passed through the channel by being deformed according to the channel height according to the blood flow velocity) And there is no particular limitation. For example, the height h _c of the channel portion may be in the range of 2 to 20 um, 2 to 17 um, 2 to 15 um, 2 to 12 um, 5 to 20 um, 5 to 17 um, 5 to 15 um, 5 to 12 um, 7 to 20 um, , 7 to 15 um, 7 to 12 um, 10 to 20 um, 10 to 17 um, 10 to 15 um, or 10 to 12 um, but is not limited thereto.

The width (w _c ) of the channel portion means an interval between adjacent ridge portions, and the widths w _c of one or more channel portions may be equal to or different from each other. The width (w _c ) of the channel portion is not particularly limited and may be suitably selected in the range of the width (w _r ) of the raised space described above, for example, but is not limited thereto.

In addition, the length? In the oblique direction of the raised space is not particularly limited, but can be adjusted according to the size of the body portion and / or the number of raised spaces formed discontinuously in one ridge portion. In another aspect, the number of discontinuous raised spaces may be adjusted according to the size of the body portion and / or the length in the oblique direction of the raised space. The lengths? In the oblique direction of two or more raised spaces formed in one ridge may be equal to or different from each other. In one example, the length delta in the oblique direction of the raised space may be at least 2 times, at least 2.5 times, at least 3 times, at least 3.5 times, at least 4 times, or at least 4.5 times the average diameter of erythrocytes, 2 to 20 times, 2.5 to 20 times, 3 to 20 times, 3.5 to 20 times, 4 to 20 times, 4.5 to 20 times, 2 to 15 times, 2.5 times 5 to 15 times, 3 to 15 times, 3.5 to 15 times, 4 to 15 times, 4.5 to 15 times, 2 to 10 times, 2.5 to 10 times, 3 to 10 times, 3.5 to 10 Fold, 4- to 10-fold, 4.5- to 10-fold, 2- to 9-fold, 2.5- to 9-fold, 3- to 9-fold, 3.5- to 9-, 4- to 9-, 2 to 8 times, 2.5 to 8 times, 3 to 8 times, 3.5 to 8 times, 4 to 8 times, 4.5 to 8 times, 2 to 7 times, 2.5 to 7 times, 3 times To 7 times, 3.5 times to 7 times, 4 times to 7 times, 4.5 times to 7 times, 2 times to 6 times, 2.5 times to 6 times, 3 times 4 to 6 times, 4.5 to 6 times, 2 to 5 times, 2.5 to 5 times, 3 to 5 times, 3.5 to 5 times, 4 to 5 times, Fold, or from 4.5 times to 5 times, but not limited thereto.

The spacing g between the raised spaces at one ridge, that is, the length of the structure (the width of the structure determined by the width of the ridge) that disconnects the ridge internal space, can be physically divided into two or more spaces And the upper limit value is not particularly limited, but it may be shorter than the length of the protruding space in consideration of the plasma separation efficiency. In one example, the spacing g between the raised spaces is in the range of 0.01 to 100um, 0.01 to 90um, 0.01 to 80um, 0.01 to 70um, 0.01 to 60um, 0.01 to 50um, 0.01 to 45um, 0.01 to 40um, 0.01 to 35um, 0.01 0.1 to 80um, 0.1 to 70um, 0.1 to 60um, 0.1 to 50um, 0.1 to 45um, 0.1 to 50um, and 0.01 to 20um, 0.01 to 20um, 0.01 to 15um, 0.01 to 12um, 0.1 to 40um, 0.1 to 35um, 0.1 to 30um, 0.1 to 25um, 0.1 to 20um, 0.1 to 15um, 0.1 to 12um, 0.1 to 10um, 1 to 100um, 1 to 90um, 1 to 80um, 1 to 70um, 1 But are not limited to, from 1 to 50, 1 to 50, 1 to 45, 1 to 40, 1 to 35, 1 to 30, 1 to 25, 1 to 20, 1 to 15, It is not.

The inflow section 301 may include an inflow port through which blood is introduced and supplied to the body section, or may have an inflow port through which blood flows. The inlet may be connected directly to the body part or may be connected to the body part through a cavity including an empty space optionally communicating with the inlet part and the body part. The inflow portion of the blood may be located at the upper portion of the body portion, and the upper portion thereof refers to an upstream region where blood flows in the body portion.

One side portion where the blood cell discharging portion 401 is located is a side surface of the body portion having an angle greater than 0 DEG and less than 90 DEG (an angle formed by the ridge portion and one side surface 400 in the moving direction of the blood) A bottom portion connected to the side surface 400, or a corner portion connected to the bottom surface of the side surface 400. In addition, the other side portion where the plasma discharging portion 501 is located is the opposite side of the side surface on which the blood cell discharging portion is located, the bottom portion connected to the side surface, or the corner portion connected to the side surface and the bottom surface portion.

The blood discharge unit and the plasma discharge unit may be connected to the body part through a cavity directly connected to the body part or optionally including an empty space communicating the discharge part and the body part.

The side surface refers to both sides based on the direction in which the blood moves in the body part. In Figs. 11 and 12, there is illustrated an example in which one side of an inclined protrusion (inclined structure) and an angle of more than 0 degrees and less than 90 degrees is right on the basis of the blood flow direction, but one side is opposite to the illustrated direction , That is, the left side with respect to the blood proceeding direction, the ridge may have an inclined structure such that the angle formed between the left side surface and the blood running direction is more than 0 degrees and less than 90 degrees.

In one example, the plasma-separating chip is a chip for separating plasma as described above, wherein the surface of the concavo-convex portion of the body portion (the ridge portion (continuous structure: 102; discontinuous structure: 105), the channel portion 103, (Fixed) the structure 100 that cuts both side surfaces or both sides of the structure 100 including both sides (including the structure 104) and the inner space of the protrusions, to the substrate 200 .

At this time, the structure 100 and the substrate 200 may be made of the same solid or different materials, and the specific material thereof is not particularly limited. The structure 100 and the substrate 200 may be formed of a material such as polystyrene (PS), polycarbonate (PC), polymethylmethacrylate (PMMA), polyethylene terephthalate (PET), poly Polymers commonly used such as polydimethylsiloxane (PDMS) and the like; SU-8, PEG-DA (polyethylene glycol diacrylate); Metals such as aluminum, iron, platinum, and copper; Soft solids such as silicon; Glass, or the like, but is not limited thereto.

The size of the plasma separation chip is not particularly limited and may be, for example, about 1 mm to about 100 mm, about 1 mm to about 50 mm, about 1 mm to about 30 mm, about 1 mm to about 20 mm, From about 100 um to about 1000 um, from about 300 um to about 2000 um, from about 300 um to about 2000 um, from about 100 um to about 2000 um, from about 100 um to about 1800 um, from about 100 um to about 1500 um, from about 100 um to about 1300 um, um to about 1800 um, from about 300 um to about 1500 um, from about 300 um to about 1300 um, or from about 300 um to about 1000 um.

Another example provides a chip array for plasma separation comprising two or more chips for plasma separation. The two or more plasma separation chips may be, for example, 2 to 10, 2 to 7, 2 to 5, 2 to 4, 2 or 3, 3 to 10, 3 to 7, Five, or three or four plasma separation chips may be connected in series. At least one of the two or more plasma separation chips may be a plasma separation chip having the discontinuous raised portion described above (see FIG. 12). In another example, in addition to the plasma separation chip (FIG. 12) having the discontinuous ridge portion, the plasma separation chip array may include one or more plasma separation chips (FIG. 11) having continuous ridges, (In the direction of the blood injection portion) of the chip for plasma separation having the ridge portion. By including the continuous ridges in the vicinity of the blood injection unit, it is possible to prevent blood clumping that may occur during blood flow in the plasma separation chip having a discontinuous ridge and clogging thereof.

In one embodiment, the plasma separation chip array comprises

(1) a blood injector into which blood is injected;

(2) a first plasma separation section connected to the blood injection section (1)

(3) a second plasma separator connected to the first plasma separator; And

(4)

. ≪ / RTI >

The first plasma separation unit is connected to the blood injecting unit so that the blood injected thereinto flows,

(i-1), a body portion in which a space through which blood can flow is integrally formed, and a channel portion and at least one ridge portion are alternately and continuously formed on an outer surface of the body portion;

(ii-1) an inflow portion formed at an upper portion of the body portion and into which blood flows;

(iii-1) a blood cell outlet formed on one side of the body; And

(iv-1) a plasma discharge portion formed on the other side surface portion of the body,

Wherein the ridge portion and the channel portion communicate with each other in the closed space,

Wherein the protruding portion is inclined at an angle of more than 0 DEG and less than 90 DEG with one side of the body portion while forming a protruding space communicating with the channel portion,

The inclined raised portion is formed continuously in an inclined direction (that is, there is no disconnection of the raised space)

One side portion on which the blood cell discharging portion is located may have a side surface and a bottom surface portion connected to the side surface of the body portion having an angle greater than 0 DEG and less than 90 DEG (an angle formed by the ridge portion and one side surface in the moving direction of the blood) Wherein the other side surface portion on which the blood plasma discharging portion is located is a side surface opposite to the one side surface on which the blood cell discharging portion is located and a bottom surface portion connected to the side surface or a corner portion where the side surface and the bottom surface portion are connected, And may include one or more chips for separating plasma containing a protruding portion.

The second plasma separation unit is connected to the plasma discharge unit of the plasma separation chip included in the first plasma separation unit so as to allow the plasma discharged therefrom to flow thereinto, and a fluid chip for plasma separation including the discontinuous ridge as described above Or more.

In one example, the first plasma separation unit may include one fluidic chip for plasma separation including the continuous protrusions, and the second plasma separation unit may include at least two plasma separation chips including the discontinuous protrusions, , ≪ / RTI >

Another example provides a plasma separation apparatus comprising one or more, for example, two or more, four or more, six or more, or eight or more of the above-described chip array for plasma separation. The upper limit value of the number of chip arrays included in the plasma separating apparatus is not limited and may be included in a permissible number of spatial conditions of the plasma separating apparatus. 1 to 100, 1 to 80, 1 to 60, 1 to 50, 1 to 40, 1 to 30, 1 to 20, and 1, respectively, of the number of chip arrays included in the plasma separating apparatus To 10, 2 to 100, 2 to 80, 2 to 60, 2 to 50, 2 to 40, 2 to 30, 2 to 20, or 2 to 10, no. Blood which can be treated in one plasma separating apparatus increases proportionally in proportion to the number of the plasma separating chip arrays included in the plasma separating apparatus.

In one example, the first plasma separation part included in each plasma separation chip array of the plasma separation device may include at least one plasma separation chip including the continuous protrusion, for example, 1 to 10, 1 to 5 1 to 3, or 1, and the second plasma separation section comprises at least one or more than two, for example, 1 to 30, 1 to 25, and 1, 1 to 10, 1 to 8, 1 to 5, 1 to 3, 2 to 30, 2 to 25, 2 to 20, 2 to 15, 2 To 10, 2 to 8, 2 to 5, 2 to 3, or 2. The two or more plasma separation chip arrays included in the plasma separation apparatus may be connected in parallel.

In another example, the plasma separation device may further include a blood cell reservoir for storing blood cells discharged from two or more chip separation arrays for plasma separation. The stored and / or separated blood cells can be reused for a desired use. In addition, the plasma separation device may further include a plasma storage part for collecting and / or storing plasma discharged from two or more plasma array chips for plasma separation.

The plasma separation chip, the plasma separation chip array, and / or the plasma separation device may be used so that blood flows from the injection part to the opposite surface due to the flow rate of the blood. It is sufficient that the infusion rate of the blood is equal to or higher than the minimum flow rate causing the flow of blood. For example, the infusion rate of blood per unit plasma array for plasma separation may be at least about 10 ul / min, at least about 20 ul / min, at least about 30 ul / min, at least about 40 ul / min, at least about 50 ul / min Min, about 800 μl / min, about 700 μl / min, more than about 60 μl / min, not less than 70 μl / min, or about 80 μl / min, about 600 μl / min, about 500 μl / min, about 450 μl / min, about 400 μl / min, about 350 μl / min, about 300 μl / About 150 ul / min, or about 120 ul / min, but is not limited thereto. The blood infusion rate of the plasma separation apparatus may be set to increase in proportion to the blood infusion rate of the plasma separation chip array according to the number of included chip separation arrays.

In one example, the plasma separation chip, the plasma separation chip array, and / or the plasma separation device may be used in connection with a supply part capable of supplying and / or injecting blood into the blood inflow part or the blood injection part.

In another example, the blood-injecting unit or the blood-introducing unit of the plasma separation chip, the plasma separation chip array, and / or the plasma separation device may further include the plasma separation chip, the plasma separation chip array, and / And a blood supply part connected to the blood supply part. The blood supply part may function to apply an injection flow rate of blood by storing blood and / or air compression, and may be a pump such as a syringe, a pipette, a piston pump, a syringe pump, a diaphragm pump, And the like. In one example, the blood injection portion of the plasma separation device may be used in connection with a syringe needle (see FIG. 10).

Another example provides a plasma separation method using the plasma separation chip, the chip array for plasma separation, and / or the plasma separation apparatus.

Specifically, in the plasma separation method,

Injecting blood into the plasma separation chip, the plasma separation chip array, and / or the plasma separation device; And

Collecting the plasma discharged from the plasma separation chip, the plasma separation chip array, and / or the plasma separation device.

The plasma separation method has an advantage that plasma can be separated efficiently from whole blood or blood having a relatively high blood cell concentration. In one example, the injected blood may be whole blood or blood diluted 1 to 20 times by volume on a whole volume basis, but is not limited thereto.

According to the plasma separation method, the separation purity (number of erythrocytes / number of injected red blood cells) is about 90% or more, about 93% or more, about 94% or more, about 95% About 95% or more, about 96% or more, about 96.5% or more, about 97% or more, about 97.5% or more, about 98% or more, about 98.5% or more or about 99% or more or about 99.5% or more. The purity of the separated plasma may be related to the amount of blood to be injected (for example, the smaller the amount of blood injected, the higher the plasma purity may be). In order to increase the efficiency of plasma separation, Min, from about 10 to about 700 ul / min, from about 10 to about 600 ul / min, from about 10 to about 1000 ul / min, from about 10 to about 1000 ul / min, from about 10 to about 1000 ul / min, Min, from about 10 to about 250ul / min, from about 10 to about 200ul / min, from about 10 to about 500ul / min, from about 10 to about 400ul / min, from about 10 to about 350ul / Min, about 50 to about 700 ul / min, about 50 to about 600 ul / min, about 50 to about 1000 ul / min, about 50 to about 1000 ul / min, about 50 to about 900 ul / Min, about 50 to about 200 ul / min, about 50 to about 200 ul / min, about 50 to about 350 ul / min, about 50 to about 350 ul / min, It can be adjusted from about 50 to about 150ul / min range range.

In addition, the plasma recovery rate (volume of collected plasma / volume of whole blood injected) by the plasma separation method may be about 5 to about 15%.

The blood separation chip, the chip separation array for blood separation, the blood separation device and / or the blood separation method using the same, which are provided in this specification, can rapidly separate plasma with high efficiency with a simpler method.

1 schematically illustrates a chip for blood separation according to an embodiment of the present invention. The structure of a chip having a continuous slant array (CSA) The right side of the figure shows the chip structure with the discrete slant array (DSA) ridges formed by the micro-posts and the movement of the blood cells. The lower left shows the chip structure including the CSA ridge (W _r : elevation space width, h _r : elevation space height, h: channel height, g: elevation space spacing), and the right side of the bottom shows the hemocyte movements in the chip section including the DSA elevation. δ: Rise space length; Δδ: Ridge interval; Dotted arrow: Intended blood cell movement path; Solid arrow: Deviated blood cell movement path).
FIG. 2 schematically shows three different types of plasma-separating chip arrays. In Type 1, three chips (first chip to third chip) having CSA ridge portions are connected in parallel, and Type 2 is a DSA (First chip to third chip) having a bulge portion are connected in parallel, Type 3 is a chip (first chip) having a CSA bulge and two chips having a DSA bulge 3 chips) are connected in parallel in this order (r1-r6 denote the division of each marked position).
Figure 3 is a bright-field micrograph showing the CSA ridge and the DSA ridge (Scale bars: 100 um).
4 and 5 show the result of testing the channel clogging phenomenon. FIG. 4 is a photograph showing the result of fluorescent staining of blood clots in a plasma separating apparatus each including a chip array of Type 1-3 , With discontinuous ridges, shows a higher rate of thrombus formation in proportion to time (Scale bars: 200 μm), as compared to the case with continuous ridges.
FIG. 5 is a graph showing the result (fluorescence intensity) of the thrombus fluorescence staining obtained in FIG. 4 according to time (blood injection flow rate: 100 uL / min).
Figure 6 is a photograph showing undesired deviation of RBCs deviating from the intended RBC stream in type 1 and type 3 plasma separation arrays, (In each photograph, the direction of blood flow is from upper to lower direction, injection flow rate is 100 μL / min, Scale bars: 200 μm).
FIG. 7 is a graph showing the purity of plasma separated in the type 1 and type 3 plasma separation apparatuses according to the flow rate ( n = 3).
FIG. 8 is a photograph exemplarily showing a device for high throughput separation of plasma, wherein red blood cells discharged from each chip array are stored in a common storage.
9 is a photograph exemplarily showing operation of a device (including eight chip arrays) in the form of a device connected to a syringe pipette (plasma flow rate: 904.3 ± 15.6 uL / min; extraction speed (min. ) Purity of plasma separated at 51 ± 1 uL / min: 99.88 ± 0.01% ( n = 3)).
Fig. 10 shows an example in which the plasma separation device is applied to a syringe pipette.
Fig. 11 is a schematic diagram showing an example of a chip for plasma separation having a continuous protrusion (inclined structure).
12 is a schematic diagram showing an example of a chip for plasma separation having a discontinuous raised body (inclined structure).

BRIEF DESCRIPTION OF THE DRAWINGS The above and other objects, features and advantages of the present invention will become more apparent from the following detailed description of the present invention when taken in conjunction with the accompanying drawings. First, the terminology used in the present application is used only to describe a specific embodiment, and is not intended to limit the present invention, and the singular expressions may include plural expressions unless the context clearly indicates otherwise. Also, in this specification, the terms "comprises" or "having" and the like are used to specify that there is a feature, a number, a step, an operation, an element, a component or a combination thereof described in the specification, But do not preclude the presence or addition of features, numbers, steps, operations, components, parts, or combinations thereof. Also, in the present specification, the numerical range expressed by "A to B" means all numerical values (real numbers) between A and B including A and B, meaning including an approximation of A and B . ≪ / RTI >

Hereinafter, the present invention will be described in more detail with reference to Examples. It is to be understood by those skilled in the art that these embodiments are only for describing the present invention in more detail and that the scope of the present invention is not limited by these embodiments.

Example  1: Design and Fabrication of Microfluidic Chip for Plasma Separation

PDMS (polydimethylsiloxane, Dow Corning, USA) was prepared. The master mold for making the PDMS chip was fabricated using photolithography. Since this chip has two steps of height, the coating of photoresist (SU-8, Microchem, USA), mask alignment, ultraviolet exposure, and development process was repeated twice on a silicon wafer. The prepared master mold was treated with silane (trichloro silane, Sigma Aldrich, USA) to facilitate the release of PDMS. PDMS mixed with base and curing agent 10: 1 was poured into the silane-treated master mold, the bubbles were removed, and the mixture was heated and cured. The fully cured PDMS was removed from the master mold and holes were punched in the inlet and outlet, and bonded to the glass via oxygen plasma treatment. The channel height of the chip was 12 μm, which is the height (hc), the width (wr) of the continuous gradient structure (CSA) was 15 μm and the slope structure height (hr) was 15 μm. In the case of the Discrete Inclined Structure (DSA), the height of the channel portion is 12 μm, the width (wr) is 15 μm, the height of the inclined structure (hr) is 15 μm and the microposts, And the slope length (d) of each of the sloped sloped structures was set to 160 [mu] m. Each CSA or DSA has a width of 1 cm and a length of 1.7 cm (3 parallel connections of about 5 cm). As shown in the lower part of FIG. 1, the discrete inclined structure (DSA) which is broken due to the micropillar structure can minimize red blood cell dispersion and can have a beneficial effect for obtaining pure plasma. A 1 mL pipette tip was inserted into a 60 mL syringe for hand held drive of the microfluidic chip.

Example  2: Plasma separation test using microfluidic chip for plasma separation

Sample Preparation

Dog blood and human blood used in the experiment were purchased from Korea Animal Blood Center and Korean Red Cross, respectively. Hematocrit was measured by measuring the length of red blood cells after centrifugation after blood was contained in the capillary. In the case of blood clots, SYTO 13 (Thermo Fisher Scientific, USA), a fluorescent material capable of staining nuclei of white blood cells in blood clots, was analyzed. The plasma separation efficiency was measured using a hematocytometer (Invitrogen, USA).

Experimental conditions

A syringe pump (KD Scientific, USA) was used to measure the plasma separation efficiency of the chips according to the flow conditions (100 to 400 uL / min). After the optimal flow rate was determined, a 1 ml pipette The experiment was carried out. A CCD camera (Nikon, Japan) and a high-speed camera (Vision Research, USA) were attached to a fluorescent microscope (Nikon) to observe the movement of blood cells in the chip. Analysis of fluorescence images was performed using image analysis software ImageJ (National Institutes of Health, USA).

result

The flow of the fluid in the inclined structure for separating blood cells flows in a direction parallel to the inclined structure and the portion below the inclined structure generates a flow of fluid toward the opposite side (direction in which the blood cells gather). At this time, when the cell size is sufficiently large (d ≥ d is the cell diameter), the force is applied to the lower side of the inclined structure, and in the direction perpendicular to the inclined structure (direction in which the blood cells gather) . The number of red blood cells in the blood is inevitably present due to the excessive number of cells that are pushed into the sloping structure and the pushed up red blood cells move along the slope structure (lead to undesired deviation along the deviation path) . We have developed a discrete slope structure to improve the separation efficiency of blood cells by minimizing this phenomenon and to improve the amount and purity of recovered plasma. The DSA (discontinuous ridge) is an array with a slope of 160um and the slope is separated by a micropost of 30um width (104 in Fig. 12). Each slant structure of the DSA was manufactured by shifting by the delta delta in the immediately preceding slant structure (see the right side of Fig. 1). The inclined structure shifted by Δδ serves to keep the movement of the hemocyte in the focusing path (the direction in which the hemocytes gather, the vertical direction of the tilt structure). The micropost between the tilted structures is pushed up by the tilted structure by the hemocytes that are focused along the focusing path, and the hemocytes moving to the opposite side can be moved back to the focusing path. This functioning DSA can maximize the purity of isolated plasma.

1 shows a plasma separation process in a plasma separation chip (right side) having a plasma separation chip (left side) having a continuous inclined structure (raised portion) and a discontinuous (separated) Show them as enemies. The discontinuous inclined structure can prevent the blood vessels from migrating to the opposite side due to the micropost between the inclined structures, thereby preventing the dispersion of blood cells and maximizing the separation efficiency. FIG. 2 schematically shows three types of plasma array chips for combination of CSA and DSA (Type 1: three CSAs (continuous gradient structure), Type 2: three DSAs; Type 3: One CSA and two DSAs). Figure 3 is a micrograph of CSA and DSA.

The channel clogging phenomenon in the case of separating blood using the three types of chip separation chips shown in FIG. 2 was tested. The clogging of the channel proceeded by fluorescence staining of blood clots. The injection volume of blood was tested at 100 uL / min. The obtained results are shown in Figs. 4 and 5. Fig. FIG. 4 shows the fluorescence images obtained. In the case of a Type 2 chip array containing only DSA, a blood clot is caught in the micropost, and the channel clogging phenomenon is conspicuous. This phenomenon becomes more prolonged (blood flows) And the phenomenon becomes worse. The change of the obtained fluorescence intensity is shown graphically in Fig. As in FIG. 4, in FIG. 5, it can be seen that, in the case of the Type 2 chip array including only the DSA, the clogging phenomenon deteriorates with time.

As can be seen from FIGS. 4 and 5, although DSA has better plasma separation efficiency than CSA, CSA has no clogging due to lack of blood clotting, whereas DSA has microposts arranged at intervals of about 100 μm, So that at least the first compartment (chip) is divided into two or more compartments (for example, three compartments) so as to separate the plasma with high efficiency without channel clogging, so that at least the first compartment It may be advantageous to have DSA included.

FIG. 6 shows a comparison of the plasma separation efficiency between the type 1 chip array and the type 3 chip array. In the case of type 1 comprising all the compartments of CSA, the plasma separation efficiency is low due to the large amount of blood cells migrating to the opposite side in the inclined structure. And the remaining two channels are composed of DSA. In the case of type 3, the plasma purity of the final recovered part is very excellent because there is no blood moving in the opposite direction while taking the inclined structure. The blood was tested at a flow rate of 100 uL / min.

7 is a graph showing the purity of plasma separated through the type 1 and type 3 chip arrays according to the flow rate. FIG. 7 also shows that the type 3 chip array is superior to the type 1 chip array in plasma purity.

Figures 8 and 9 show that the amount of plasma separation can be increased by including the chip array in parallel. In other words, while one chip array (Type 3; three chips (1 CSA + 2 DSA)) can process approximately 100 uL / min, a blood separator including eight chip arrays in parallel ), It is possible to treat 904.3 ± 15.6 uL of blood per minute (see FIG. 9).

The above-described embodiments are illustrative in all aspects and do not limit the scope of the present invention.

100: concave-convex forming face and both sides of the body part
101: Body part
102: continuous ridge
103:
104: Structure that cuts the internal space of the ridge
105: discontinuous ridge
200: opposite surface of the concave-convex forming surface of the body part
301: blood inflow section
401: blood cell outlet
501: Plasma outlet part

Claims (23)

(i) a body portion in which a space through which blood can flow is integrally formed, and a channel portion and a ridge are alternately and continuously formed on one surface;
(ii) an inflow portion through which the blood located above the body portion flows;
(iii) a blood-collecting part located at one side of the body part; And
(iv) a plasma outlet part located on the other side of the body part
Including the
The ridge portion and the channel portion communicate with each other in a space where the blood can flow,
Wherein the protruding portion has an inner space having a height equal to the height of the channel portion and the height of the protruding space formed by forming a protruding space communicating with the channel portion and inclined at an angle of more than 0 DEG and less than 90 DEG with one side surface of the body portion,
Wherein the inclined raised portion is discontinuously formed by at least one structure that cuts the inner space in an inclined direction,
One side portion where the blood cell discharging portion is located is a side portion of the body portion having an angle of less than 90 degrees with the inclined surface of the ridge portion, a bottom portion connected to the side surface, or a corner portion where the side surface-
Wherein the other side surface portion on which the plasma discharge portion is located is formed on an opposite side surface of one side surface on which the blood cell discharge portion is located, a bottom surface portion connected to the side surface,
Chip for plasma separation. delete The plasma separation chip according to claim 1, wherein the protrusions are formed to be inclined at an angle of 45 ° or more and less than 90 ° with one side of the body. The plasma separation chip according to claim 1, wherein a height (hr) of the protruding space formed in the protruding portion is 1 to 20 um. The plasma separation chip according to claim 4, wherein a width of the protruding space formed in the protruding portion is 0.5 to 3 times the height of the protruding space. The plasma separation chip according to claim 1, wherein a length of the ridge portion of the ridge space formed in the ridge portion in an oblique direction is 2 to 20 times the diameter of a red blood cell. The plasma separation chip according to claim 1, wherein the channel portion has a height of 2 to 20 um. (1) a blood injector into which blood is injected;
(2) a first plasma separation section connected to the blood injection section (1)
(3) a second plasma separator connected to the first plasma separator; And
(4)
Lt; / RTI >
The first plasma separation unit is connected to the blood injecting unit so that the blood injected thereinto flows,
a body part formed integrally with a space through which blood can flow in the inner part (i-1) and alternately forming a channel part and a ridge alternately on one surface; (ii-1) an inflow portion formed at an upper portion of the body portion and into which blood flows; (iii-1) a blood cell outlet formed on one side of the body part; And (iv-1) a plasma discharging portion formed on the other side surface portion of the body portion, wherein the ridge portion and the channel portion communicate with each other in a space where the blood can flow, and the ridge portion forms a ridge space communicating with the channel portion Wherein the inclined protruding portion is inclined at an angle of more than 0 ° and less than 90 ° with one side surface of the body portion, the inclined protruding portion being continuously formed in an inclined direction, Wherein one side portion where the blood cell discharging portion is located is a side surface of a body having an angle of less than 90 DEG with respect to an inclined surface in a direction in which the blood of the protruding portion is moved, a bottom portion connected to the side surface, The other side surface portion on which the plasma discharging portion is positioned is provided on the opposite side surface of one side on which the blood cell discharging portion is located, And a continuous ridge portion in which the side surface and the bottom surface portion are connected to each other,
Wherein the second plasma separation unit is connected to the plasma discharge unit of the plasma separation chip included in the first plasma separation unit so that the plasma discharged therefrom is introduced into the second plasma separation unit, And at least one chip for plasma separation comprising a ridge.
Chip array for plasma separation. 9. The plasma separation apparatus according to claim 8, wherein the first plasma separation unit comprises at least one plasma separation chip including the continuous protrusions, and the second plasma separation unit comprises at least two plasma separation chips including the discontinuous protrusions, Wherein the plasma separation chip array comprises: A plasma separation device comprising at least one chip array for plasma separation according to claim 8. 11. The plasma processing apparatus according to claim 10, wherein the first plasma separation unit included in each of the plasma separation chip arrays includes at least one plasma separation chip including the continuous protrusions, and the second plasma separation unit includes the discontinuous protrusions A plasma separation device comprising two or more chips for plasma separation. 9. A method for separating blood plasma, comprising: injecting blood into the plasma separation chip according to any one of claims 1 to 7; And
Collecting the plasma discharged from the plasma separation chip
≪ / RTI > 13. The plasma separation method according to claim 12, wherein the blood to be injected is whole blood or whole blood diluted 1 to 20 times by volume. 10. A method for separating blood plasma, comprising: injecting blood into the plasma separation chip array of claim 8; And
Collecting plasma discharged from the plasma separation device
≪ / RTI > 15. The plasma separation method according to claim 14, wherein the injected blood is whole blood or whole blood diluted 1 to 20 times by volume. 10. A method for treating blood plasma, comprising: injecting blood into the plasma separation apparatus of claim 10; And
Collecting plasma discharged from the plasma separation device
≪ / RTI > 17. The plasma separation method according to claim 16, wherein the injected blood is whole blood or whole blood diluted 1 to 20 times by volume. 12. A method for treating blood plasma, comprising: injecting blood into the plasma separation apparatus of claim 11; And
Collecting plasma discharged from the plasma separation device
≪ / RTI > 19. The method of claim 18, wherein the injected blood is whole blood or whole blood diluted 1 to 20 times by volume. A chip array for blood separation according to claim 8; And
A blood supply part connected to the blood injection part of the blood-
And a blood collection port. 21. The blood separation kit according to claim 20, wherein the blood supply part is selected from the group consisting of a syringe, a pipette, a piston pump, a syringe pump, a diaphragm pump, and a tube peristaltic pump. A blood separation device according to claim 10; And
A blood supply unit connected to the blood injection unit of the blood separator,
And a blood collection port. 23. The blood separation kit according to claim 22, wherein the blood supply unit is selected from the group consisting of a syringe, a pipette, a piston pump, a syringe pump, a diaphragm pump, and a tube peristaltic pump.

Images