KR101248750B1 - Biosensor comprising embossed capillary channels - Google Patents

Abstract

The present invention provides an optical biosensor comprising an embossed capillary channel to quickly and accurately measure a desired result via biomaterial through color intensity. According to the present invention, it is possible to provide a biosensor capable of quick and accurate measurement and at the same time having a low manufacturing cost.

Description

Biosensor comprising embossed capillary channels

The present invention relates to a biosensor. More specifically, the present invention relates to an optical biosensor comprising an embossed capillary channel to quickly and accurately measure a predetermined result via a biomaterial through color change.

Biosensor immobilizes biomaterials (enzymes, antigen-antibodies, ligands, DNA, etc.) that can react by specific / selective binding to specific substances to specific mediators to produce the desired amount of light, electrochemistry, It refers to any system that converts and measures various signals such as fluorescence, surface plasmon resonance (SPR), field-effect transistor (FET), quartz crystal microbalance (QCM), and heat generation.

Biosensors can be classified into optical biosensors using light recognition and electrochemical biosensors using recognition using electrical values.

Here, in the case of the optical biosensor, an optical device that can sense light, a mineral that can cause color development, and an external device that can measure the change or intensity of such light without external interference are important parts. Blood glucose measurement for diabetics is a large part of the photobiosensor market, but in addition to blood sugar, cholesterol, hemoglobin, glycosylated hemoglobin (HbA1c), lactic acid, and other diagnostically important clinical proteins can be measured. Optical biosensors are also commercially available. In particular, the optical biosensor is operated by the principle of sensing the numerical value to be measured by the optical element by detecting the light when the sample is injected into the color of the sample reaction section chemically treated with minerals, the color is irradiated.

An example of such a conventional optical biosensor is disclosed in US Pat. No. 5,418,142, as shown in FIG. Samples such as blood are introduced through the inlet (2) of the kit (1), reacted and developed in the sample reaction unit (3), and then the kit (1) is put into a separate optical device (not shown). The degree of numerical value to be measured is confirmed by checking the degree of color change of the optical element in the optical device.

The conventional optical biosensor as shown in FIG. 1 has the following problems.

First, red blood cells in a sample, such as blood, can affect color development and cause inaccurate results. One way to solve this problem is to correct the wavelength of the irradiated light, but the device is complicated and expensive to manufacture.

Secondly, when a sample such as blood is supplied in excess and all the supplied samples have a color reaction, different results may be produced than when the same sample is supplied in a small amount. That is, there is a problem that inaccurate measurements are made in which the results vary depending on the amount of sample supplied.

Third, in the case of the conventional optical biosensor, it takes a few minutes before the sample is introduced to reach the sample reaction unit and react. Significant time from the input of the sample to the acquisition of the reaction result can pose a serious risk for the patient, especially in the procedure using the biosensor directly.

The present invention has been made to solve the above problems.

That is, to prevent the red blood cells in the sample such as blood from affecting the color development, and even if the sample is excessively controlled so that only an appropriate amount can be reacted, so as to propose a biosensor that can produce accurate results in any case do.

In addition, the present invention proposes a biosensor capable of quickly performing all procedures from inflow of blood to obtaining results.

In addition, when a plurality of channels are located inside the biosensor, it is proposed a biosensor that can lead to accurate results by effectively releasing the air existing in the channel to the outside before sample inflow.

At the same time, we want to propose a biosensor that can reduce manufacturing costs by eliminating the need for special chemical treatment or complicated equipment.

In order to solve the above problems, an embodiment of the present invention, the carrier layer; An analyte detection layer positioned on a portion of the carrier layer and including a sample reaction part; And a cover layer positioned on the carrier layer and the analyte detection layer, the cover layer including an injection hole into which a sample is injected, the cover layer being in communication with the injection hole and disposed laterally with the analyte detection layer. And a channel, wherein said embossed capillary channel is in fluid communication with said analyte detection layer transversely.

Another embodiment of the invention, the carrier layer; An analyte detection layer positioned on a portion of the carrier layer and including a sample reaction part; And a cover layer positioned on the carrier layer and the analyte detection layer, the cover layer including an injection hole into which a sample is injected, the cover layer being in communication with the injection hole and disposed laterally with the analyte detection layer. A channel, and a gap extending between the embossed capillary channel and the analyte detection layer and extending in a direction perpendicular to or crossing the embossed capillary channel while communicating with the outside air. Characterized in that, it provides a biosensor.

Here, the gap is preferably formed between the cover layer and the carrier layer.

In addition, it is preferable that an adhesive layer is located between the cover layer and the analyte detection layer and / or between the cover layer and the carrier layer.

In addition, the vent hole is preferably formed in the cover layer on the analyte detection layer.

In addition, a spacer is preferably located between the carrier layer and the cover layer.

In addition, the sample is preferably changed in color on the analyte detection layer.

In addition, the analyte detection layer preferably includes an erythrocyte separation unit located between the sample reaction unit and the embossed capillary channel.

In addition, a capillary channel is preferably formed between the cover layer and the analyte detection layer.

In addition, the carrier layer is preferably circular, square or strip type.

In addition, the irradiation opening in the region corresponding to the sample reaction portion in the carrier layer is preferably located.

By the above problem solving means, it is possible to provide a biosensor capable of fast and accurate measurement and at the same time low manufacturing cost.

1 is a perspective view showing a biosensor according to the prior art.
2 is a perspective view showing a biosensor according to the first, second and fourth embodiments of the present invention.
3A to 3C show, respectively, an exploded perspective view, a side view, and a cross-sectional view of a biosensor according to a first embodiment of the present invention.
4A to 4C show, respectively, an exploded perspective view, a side view, and a sectional view of a biosensor according to a second embodiment of the present invention.
5A to 5C show, respectively, a perspective view, an exploded perspective view and a sectional view of a biosensor according to a third embodiment of the present invention.
6A is an exploded perspective view of a biosensor according to a fourth embodiment of the present invention, and FIG. 6B is a cross-sectional view comparing the biosensors according to the fourth embodiment of the present invention and the biosensors according to the first and second embodiments.
7A to 7E are perspective views illustrating various embodiments of the embossing and receiving portion of the biosensor according to the present invention.
8A and 8B are conceptual views illustrating the multiple capillary channels of the biosensor according to the present invention.
9 is a graph illustrating an embodiment of a biosensor according to the present invention.

Explanation of Structure of Biosensor

Hereinafter, various embodiments of the biosensor 100 according to the present invention will be described in detail with reference to the accompanying drawings. Strip type biosensor 100 is shown in the following figures, of course, any other shape that can achieve the effect of the present invention, such as circular, elliptical, square or other polygonal is possible.

≪ Embodiment 1 >

3A to 3C show a biosensor 100 according to a first embodiment of the present invention. 6B illustrates a cross-sectional view of the biosensor 100 according to the first embodiment.

The biosensor 100 according to the first embodiment of the present invention includes a cover layer 10, an analyte detection layer 20 and a carrier layer 40, and a cover layer 10 and an analyte detection layer 20. Further comprising an adhesive layer 30 between).

The cover layer 10 includes an injection hole 12 into which a sample is introduced and an embossing 11 forming an embossed capillary channel so that the sample injected through the injection hole 12 moves to the analyte detection layer 20. do. In addition, the user may further include a support 13 for placing a finger or the like for sample input.

Here, the embossing 11 has a constant volume. Therefore, even if a user inputs an excessive amount of sample, only a certain amount of sample is introduced into the embossing 11.

The embossed capillary channel communicates with the inlet 12 so that the sample injected through the inlet 12 reaches the analyte detection layer 20 through the embossed capillary channel and the gap 22 described below. . As shown in FIG. 3A, the embossed capillary channel is in transverse communication with the analyte detection layer 20.

In addition, a vent hole (not shown) may be disposed on the cover layer 10 to allow the air inside the biosensor to flow out.

It goes without saying that the shape of the embossing 11, the inlet 12, and the support 13 is not limited. 7A to 7E illustrate various embodiments of the embossing 11, the inlet 12, and the support 13, but the embossing 11 is sufficient by forming an embossed capillary channel and may be of other forms than shown. Pay attention to

The analyte detection layer 20 is located under the cover layer 10, and consists of an erythrocyte separation unit 20a and a sample reaction unit 20b in order according to the progress direction of the sample (see FIG. 8A). The analyte detection layer 20 may be formed of a porous membrane.

The sample is injected from one side of the analyte detection layer 20 through the gap 22, formed between the analyte detection layer 20 and the cover layer 10 and in contact with the analyte detection layer 20. It is moved to the other side through 35 (see (B) of FIG. 6B).

The sample first comes into contact with the red blood cell separation unit 20a of the analyte detection layer 20.

The red blood cell separation unit 20a may be subjected to a predetermined chemical treatment to separate red blood cells in the sample, thereby lowering the number of red blood cells included in the sample to a predetermined range or less. Any chemical treatment that can separate red blood cells is possible. In this manner, by lowering the number of red blood cells in the sample reacting in the sample reaction unit 20b, color development by red blood cells can be minimized to obtain color change not affected by red blood cells, resulting in accurate measurement.

Note that the red blood cell separation unit 20a is not treated with a reagent that develops color reaction with the sample.

Samples whose erythrocyte levels are lowered below a predetermined range through the erythrocyte separation unit 20a continue to go through a single capillary channel to contact the sample reaction unit 20b.

Since the sample reaction part 20b is chemically treated for the color reaction, when the sample reacts with the color reaction, light is irradiated through the irradiation opening 41 to confirm the color development.

Meanwhile, the spacer 21 functions to secure a space between the cover layer 10 and the carrier layer 40. Here, a gap 22 through which a sample may flow is formed between the analyte detection layer 20 and the spacer 21.

The gap 22 communicates with the embossed capillary channel and analyte detection layer 20 formed by the embossing 11. In particular, it intersects with the embossed capillary channel, causing the sample injected through the inlet 12 to flow quickly through the gap 22 by capillary forces.

In addition, as the gap 22 communicates with the outside air, air to be discharged to the outside according to the inflow of the sample may be discharged to the outside.

Referring to FIG. 8B, it can be seen that in the illustrated embodiment the embossed capillary channel and the gap 22 by the embossing 11 form a T-shaped gap. However, it is noted that this is only an example for description and is not limited to this T-shape.

The adhesive layer 30 is positioned between the cover layer 10, the analyte detection layer 20, and the spacer 21 to function as an adhesive. As for the adhesive layer, another adhesive layer (not shown) may be added between the cover layer 10 and the carrier layer 40.

The carrier layer 40 is positioned under the analyte detection layer 20 and spacer 21.

On the other hand, the carrier layer 40 is located below the sample reaction portion 20b is an irradiation opening 41 for irradiating light to the sample reaction portion 20b to measure the degree of color development.

≪ Embodiment 2 >

4A-4C illustrate a biosensor 100 according to a second embodiment of the present invention.

In the second embodiment, unlike the first embodiment, the spacer 21 is not located.

The gap 32 of the second embodiment performs a function in which the air inside is discharged to the outside as the sample flows in, like the gaps 22 and 32 of the first embodiment.

Third Embodiment

5A to 5C show a biosensor according to a third embodiment of the present invention.

In the biosensor according to the third embodiment, the sample is input through the injection hole 12 'located at one side of the biosensor instead of the embossing. That is, in the first and second embodiments, the user injects a sample such as blood from the finger into the inlet 12 by placing a finger on the supporting part 13, but in the third embodiment, the user inserts the sample into the inlet 12 '. The sample may be injected into the injection hole 12 ′ by placing a finger on the side or the edge of the biosensor where) is located.

The sample introduced as in the first and second embodiments proceeds to the analyte detection layer 20 'through the gap 22' by capillary force. The air, which was previously located, is discharged to the outside of the biosensor through the gap, and the sample is in wide contact with one side of the analyte detection layer 20 '. The figure shows a T-shaped gap, but any other shape is possible.

As described above, the analyte detection layer 20 'includes a red blood cell separation unit and a sample reaction unit.

In the first and second embodiments, when the volume of the embossing 11 is constant, leading to accurate measurement even when the sample is over-injected, in the third embodiment, the volume of the inlet 12 'is constant and over-injected. Only a certain amount of sample can proceed to the analyte detection layer 20 '.

Although no separate adhesive layer is shown in this embodiment, an adhesive layer may be added between the cover layer 10 'and the analyte detection layer 20' and / or between the cover layer 10 'and the carrier layer 40'. Of course it can.

<Fourth Embodiment>

6A shows a biosensor 100 according to a fourth embodiment of the invention. 6B shows the longitudinal cross-sectional view A according to the fourth embodiment in comparison with the longitudinal cross-sectional view B according to the first and second embodiments.

Unlike the first and second embodiments, the fourth embodiment does not include the adhesive layer 30.

In the first and second embodiments where the adhesive layer 30 is present, a single capillary channel between the cover layer 10 and the analyte detection layer 20 due to the adhesive layer 30 as shown in FIG. 6A (B). 35 is formed, and the sample flows into the analyte detection layer 20 by the capillary force.

In the fourth embodiment where no adhesive layer 30 is present, a plurality of capillary channels 25 of the analyte detection layer 20 itself formed of a porous membrane causes capillary forces, as shown in FIG. 6B (A). As a result, the sample rapidly flows into the analyte detection layer 20.

That is, capillary forces are caused by one or more capillary channels regardless of the presence or absence of the adhesive layer 30, and the sample is rapidly introduced into the analyte detection layer 20.

Meanwhile, in the embodiment illustrated in FIG. 6A, an embodiment including the embossing 11 and the spacer 21 but not the adhesive layer 30 is illustrated, but the spacer 21 is not included as described in the second embodiment. Of course, embodiments without including the adhesive layer 30 are also possible.

Explanation of Principle of Biosensor

Hereinafter, the principle of the biosensor 100 according to the present invention will be described with reference to FIGS. 8A and 8B. FIG. 8A illustrates an exploded view of a portion of the cover layer 10 and FIG. 8B shows each channel through which a sample flows in a cube.

For the following description, the capillary force of the first capillary channel A, which is an embossed capillary channel, a, the capillary force of the second capillary channel B, b, the capillary force of the third capillary channels C, D, Referred to as c, in the strip-shaped biosensor 100 shown in Figures 8a and 8b refers to the longitudinal direction in the first direction and the width direction in the second direction. That is, the first direction is the longitudinal direction of the T-shaped gap and the second direction is the width direction of the T-shaped gap, and it is preferable that the first direction and the second direction cross each other, and it is particularly preferable to be perpendicular as shown.

The user injects a sample such as blood through the inlet 12 by placing a finger or the like on the base 13.

Only a certain amount of the sample injected through the inlet 12 corresponding to the volume of the embossing 11 flows downward through the first capillary channel A located below the embossing 11 and in the first direction. Flow.

The second capillary channel B formed in the second direction communicates with the first capillary channel A so that the sample flows along the second capillary channel B by capillary action. For this purpose, the relation of capillary force is preferably a <b.

Meanwhile, as the sample flows along the first capillary channel (A) and the second capillary channel (B), air previously located in the first capillary channel (A) and / or the second capillary channel (B) before the sample is introduced. The biosensor 100 should be discharged to the outside. To this end, the second capillary channel B communicates with the outside air, so that air can be discharged to the outside.

The third capillary channels C and D formed in the first direction communicate with the second capillary channel B so that the sample flows along the third capillary channels C and D by capillary action. For this purpose, the relationship between the capillary forces is preferably b <c. As described above, the third capillary channels C and D may be formed of one capillary channel 35 or a plurality of capillary channels 25.

The third capillary channels C and D contact the sample reaction part 20b. Therefore, a certain amount of red blood cells in the sample introduced into the third capillary channel (C, D) is absorbed and separated by the chemically treated red blood cell separation unit 20a (C), thereby the sample containing a red blood cell value of a predetermined range Is introduced into the sample reaction unit 20b (D), the color change is made in the sample reaction unit 20b.

Subsequently, the light for measuring color intensity is irradiated to the sample reaction part 20b in which the color change is made through the irradiation opening 41 (E), and the color intensity is measured by using an optical element and the like to measure the color intensity. Is done.

Here, again examining the relationship between the capillary forces, the inlet 12 when the force by the third capillary channels (C, D) is the largest, that is, a <b <c or a <c or b <c Most preferably, the sample injected through is automatically and quickly flows through the first capillary channel (A) to the third capillary channel (C, D).

Experimental Example 1. Investigation of the proper use conditions of the biosensor

The measurement by the biosensor 100 according to the present invention uses a reflection photometer which is an external device. Reflectance values, which measure the color intensity, are converted to glucose concentration if the black curve is valid.

A. Assay curves are made by measuring multiple venous blood samples with varying glucose concentrations. Reflectance values and glucose concentrations of the venous blood sample, measured by the reference method, can be used to generate the calibration curve.

In measurement variable 1, a sample containing glucose at concentrations of 57 mg / dl, 145 mg / dl, 242 mg / dl, 370 mg / dl, 493 mg / dl and 584 mg / dl was provided to the biosensor 100 of the present invention, and time By measuring the reflectance according to, the average reflectance values of the ten biosensors 100 are shown in FIG.

In measurement variable 2, 3 μl, 5 μl, 7 μl, 10 μl and 14 μL of venous blood were provided to the biosensor 100 of the present invention, and the reflectance was measured after 10 seconds, and the individual reflectances were measured using the calibration curve. To glucose concentration. Deviations from accuracy are shown in Table 1, measured from the average concentrations of the ten test carriers and the reference values of the blood samples.

In measurement variable 3, 3 μl, 5 μl, 7 μl, 10 μl and 14 μl of venous blood were provided to the biosensor 100 of the present invention, and the reflectance was measured after 20 seconds, and the individual reflectances were measured using the calibration curve. To glucose concentration. Deviations from accuracy are shown in Table 2, measured from the mean concentrations of the ten test carriers and the reference values of the blood samples.

In variance 4, 3 μl, 5 μl, 7 μl, 10 μl and 14 μl of venous blood were provided to the biosensor 100 of the present invention, and the reflectance was measured after 30 seconds to determine the individual reflectance using the calibration curve. To glucose concentration. Deviations from accuracy are shown in Table 3, measured from the mean concentrations of the ten test carriers and the reference values of the blood samples.

B. In the case of measured variable 1, all color development was completed within 20 seconds as shown in FIG. 9, and the reflectance value was increased in a concentration-dependent manner within the concentration range of 57 mg / dl to 493 mg / dl. .

In measurement variable 2, when the biosensor of the present invention was measured 10 seconds after the reaction, it was found that the increase in reflectance appeared without any problem when using a venous blood sample in the range of 3 μl to 14 μl.

In measurement variable 3, when the biosensor of the present invention was measured 20 seconds after the reaction, it was found that the increase in reflectance appeared without any problem when using a venous blood sample in the range of 3 μl to 14 μl.

In the case of measurement variable 4, the biosensor of the present invention, when measured 30 seconds after the reaction, was found to exhibit an increase in reflectance without any problem when using a venous blood sample in the range of 3 μl to 14 μl.

Sample volume Calculated concentration according to calibration curve (mg / dl) % Deviation from reference value 3 μl 150.3 2.95 5 μl 148.2 1.51 7 μl 146.4 0.27 10 μl 144.8 -0.82 14 μl 141.3 -3.22

<Error due to test volume in measurement variable 2>

Sample volume Calculated concentration according to calibration curve (mg / dl) % Deviation from reference value 3 μl 147.2 0.82 5 μl 146.3 0.21 7 μl 145.6 -0.27 10 μl 143.1 -1.99 14 μl 139.7 -4.32

<Error due to test volume in measurement variable 3>

Sample volume Calculated concentration according to calibration curve (mg / dl) % Deviation from reference value 3 μl 147.6 1.10 5 μl 148.3 1.58 7 μl 147.4 0.96 10 μl 142.7 -2.26 14 μl 140.1 -4.04

<Error due to test volume at measurement variable 4>

Experimental Example 2 Investigation of Haematocrit Effect of Biosensor

A. In measurement variable 5, venous blood containing 20% and 60% hematocrit was provided to the biosensor 100 of the present invention, and the reflectance was measured after 10 seconds, and the individual reflectances were measured using glucose curves. Switched to Deviations from accuracy are shown in Table 4, measured from the mean concentrations of the ten test carriers and the reference values of the blood samples.

In measurement variable 6, venous blood containing 20% and 60% hematocrit was provided to the biosensor 100 of the present invention, and the reflectance was measured after 20 seconds to convert individual reflectances to glucose concentrations using the calibration curve. It was. Deviations from accuracy are shown in Table 5, measured from the mean concentrations of the ten test carriers and the reference values of the blood samples.

B. In measurement variable 5, when the biosensor of the present invention was measured 10 seconds after the reaction, it can be seen that the increase in reflectance without any problem appears when using a venous blood sample containing 30% to 60% of hematocrit. there was.

In measurement variable 6, when the biosensor of the present invention was measured 20 seconds after the reaction, it was found that an increase in reflectance appeared without any problem when using a venous blood sample containing 30% to 60% of hematocrit.

20% hematocrit blood 60% hematocrit blood Calculated concentration according to calibration curve (mg / dl) % Deviation from reference value Calculated concentration according to calibration curve (mg / dl) % Deviation from reference value 57.4 0.70 57.80 1.40 146.1 0.76 146.6 1.10 243.9 0.79 243.7 0.70 366.1 -1.05 365.3 -1.27 490.3 -0.55 486.8 -1.26

<Effect of Hematocrit on Measurement Variable 5>

20% hematocrit blood 60% hematocrit blood Calculated concentration according to calibration curve (mg / dl) % Deviation from reference value Calculated concentration according to calibration curve (mg / dl) % Deviation from reference value 57.1 0.18 58.1 1.93 145.8 0.55 147.2 1.52 242.9 0.37 244.1 0.87 367.4 -0.70 363.1 -1.86 488.9 -0.83 484.5 -1.72

<Effect of Hematocrit on Measurement Variable 6>

Although the preferred embodiments of the present invention have been described, the present invention is not limited to the specific embodiments described above. That is, those skilled in the art to which the present invention pertains can make many changes and modifications to the present invention without departing from the spirit and scope of the appended claims, and all such appropriate changes and modifications are possible. Equivalents should be considered to be within the scope of the present invention.

10: cover layer
11: embossing
12: inlet
13:
20: analyte detection layer
20a: red blood cell separation unit
20b: sample reaction part
21: spacer
30: Adhesive layer
41: irradiation opening
40: carrier layer
100: biosensor

Claims (11)

delete Carrier layer;
An analyte detection layer positioned on a portion of the carrier layer and including a sample reaction part; And
A cover layer disposed on the carrier layer and the analyte detection layer, the cover layer including an injection hole into which a sample is injected;
The cover layer comprises an embossed capillary channel in communication with the inlet and disposed laterally with the analyte detection layer, and
A gap extending between the embossed capillary channel and the analyte detection layer while communicating with the embossed capillary channel in a direction perpendicular to or intersecting with the embossed capillary channel to communicate with the outside air; doing,
Biosensor.
The method of claim 2,
The gap is formed between the cover layer and the carrier layer,
Biosensor.
The method according to claim 2 or 3,
The adhesive layer is located between the cover layer and the analyte detection layer or between the cover layer and the carrier layer,
Biosensor.
The method according to claim 2 or 3,
A vent hole is formed in the cover layer on the analyte detection layer.
Biosensor.
The method according to claim 2 or 3,
Characterized in that the spacer is located between the carrier layer and the cover layer,
Biosensor.
The method according to claim 2 or 3,
The sample is characterized in that the color change on the analyte detection layer,
Biosensor.
The method according to claim 2 or 3,
The analyte detection layer comprises an erythrocyte separation unit located between the sample reaction unit and the embossed capillary channel,
Biosensor.
The method according to claim 2 or 3,
Capillary channels are formed between the cover layer and the analyte detection layer,
Biosensor.
The method according to claim 2 or 3,
The carrier layer is characterized in that the round, square or strip (strip) type,
Biosensor.
The method according to claim 2 or 3,
Irradiation opening is located in the area corresponding to the sample reaction portion in the carrier layer,
Biosensor.

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