KR20130114509A - System for analyzing biological sample and method for analyzing biological sample using the system - Google Patents

Abstract

PURPOSE: A biological sample analysis system for disease diagnosis and an analysis method of the biological sample for the disease diagnosis using the same are provided to maximize the reaction participation amount of biological samples for disease diagnosis by using a 3D matrix structure with a fixated component for capture recognizability, thereby improving the sensitivity of the diagnosis. CONSTITUTION: A fixation apparatus for capture recognizability for biological sample analysis for disease diagnosis comprises: a column(2) with an inner space in which both ends are open; a matrix(4) placed inside the column in which a component for capture recognizability is fixated; a porous membrane(3) which is placed inside the column and contacts the upper and lower part of the matrix in which the component for capture recognizability is fixated; and a syringe pump(1) which is placed inside the column and has a set distance from the matrix and porous membrane. An analysis method of the biological sample for the disease diagnosis comprises steps of: a step of contacting one end of a fixation device for the component for capture recognizability with a sample collecting device; a step of reacting analytes inside the sample with the component for capture recognizability inside the matrix by drawing the sample in a sample collecting device into an inner column through operating the syringe pump of the fixation device of the component for capture recognizability; a step of washing unreacted components in the matrix by drawing a washing liquid from a washing device into the inner column by operating the syringe pump of the fixation device of the component for capture recognizability; and a step of detecting a conjugate of the analytes and the components reacted in the matrix by drawing a detection reagent from a detection reagent processing unit into the inner column by operating the syringe pump of the fixation device of the component for capture recognizability. [Reference numerals] (AA) Sample collecting device; (BB,DD) Washing device; (CC,EE) Detection reagent processing device

Description

Biological sample analysis system for disease diagnosis and method for analyzing biological sample using disease {System for analyzing biological sample and method for analyzing biological sample using the system}

The present invention provides a biological sample analysis system that improves the sensitivity of diagnosis by maximizing the reaction participation of a biological sample for disease diagnosis using a three-dimensional matrix structure in which a capture recognition component is immobilized, and a biological sample analysis method using the same. It is about.

Various fields, such as veterinary, defense, food and medical, have developed techniques for measuring the extent of nucleic acid changes, cellular changes and protein changes to detect the occurrence of contamination, mutations and diseases. In particular, the technique for measuring the degree of protein change is measured using a very unique and intimate binding method such as an antigen-antibody reaction, thereby increasing the specificity and affinity of the biometric reaction, making the analysis principle universal, and applying various Analytical systems have been developed.

As an example, the development of an enzyme-linked immunosolvent assay (ELISA) method that measures the presence and concentration of proteins and antibodies in a sample using multiwell microplates as an immobilization target has provided an opportunity for various diagnosis and analysis. This method prepares a surface by immobilizing a protein or antibody at a position to be immobilized first, and reacts a sample solution in which the target protein is mixed on the surface for a predetermined time, and then uses an antibody or the like that can detect the target protein. This method recognizes and quantifies the binding to the enzyme through the color reaction of the enzyme.

However, the two-dimensional fixation method as described above is difficult to fix high-density biomolecules in a small area, and also difficult to preserve the bioactive function of the biomolecules bound thereto. In addition, the amount of protein obtained from a biological sample such as blood or tissue is very small for the diagnosis of a disease of a patient, and the analysis range is 10 pg / ml to 1 ug / ml for a protein or an infectious agent (for example, a virus or a bacterium). In the case of 1 x 10 ⁵ particles / ml or more can be detected, which is 10 times less sensitive than the PCR method.

After that, the development of a high-speed diagnostic kit that predicts the presence of a corresponding antibody and protein by binding proteins and antibodies to a porous membrane, etc., and an automated diagnostic device capable of precise inspection such as a biosensor array chip and a lab-on-a-chip And supplied, but still have low sensitivity and are not used primarily for detection of infections. Recently, nano-biosensor technology, which is a fusion of biotechnology and nanotechnology, has been in the spotlight. As a result, the concept of nanosensors and immunoassay methods based on vibrating cantilevers have been reported. It exists.

As described above, the existing studies are pursuing nano-integration, integration, and simplicity, but since the detection limitations are higher than those of detection techniques such as PCR, the existing researches reveal limitations in the practical field.

Therefore, the main object of the present invention is to maximize the reaction participation of the biological sample for disease diagnosis using the three-dimensional matrix structure in which the capture recognition component is immobilized, thereby dramatically increasing the detection sensitivity of the analyte present in the sample in a small amount. A capture recognition component immobilization device for analyzing a biological sample for diagnosing a disease as a means to be obtained. And to provide a method for producing the same.

It is also an object of the present invention to provide a biological sample analysis system for diagnosing a disease comprising the capture recognition component immobilization device.

It is also an object of the present invention to provide a biological sample analysis method for disease diagnosis using a biological sample analysis system for disease diagnosis.

As one aspect for achieving the above object, the present invention

A column having both ends open and having a space therein;

A matrix present in the column and immobilized with capture components;

A porous membrane present in the column and in contact with the top and bottom of the matrix to which the capture recognition component is immobilized; And

A syringe pump present in the column and at a distance from the matrix and the porous membrane;

A capture recognition component immobilization device for analyzing biological samples for disease diagnosis.

1 in the present specification shows a preferred embodiment of the capture recognition component immobilization device. However, the present invention is not limited to the above.

As another aspect, the present invention

The capture recognition component immobilization device for diagnosing the disease;

A sample collection device;

Washing device; And

Comprising a detection reagent processing apparatus,

A biological sample analysis system for disease diagnosis.

1 in the present specification shows a preferred embodiment of the biological sample analysis system. However, the present invention is not limited to the above.

As another aspect, the present invention

Fixing the capture recognition component to the matrix;

Depositing a matrix in which the capture recognition component is immobilized in the column; And

Positioning the syringe pump in the column

The present invention relates to a method of manufacturing a capture recognition component immobilization device for disease diagnosis.

As another aspect, the present invention

Contacting one end of the capture recognition component immobilization device with the sample collection device;

Operating a syringe pump of the capture recognition component immobilization device to suck a sample present in the sample collection device into a column to react the capture recognition component in the matrix with the analyte in the sample;

Operating a syringe pump of the capture recognition component immobilization device to suck the washing solution present in the cleaning device into the column to wash components not involved in the reaction in the matrix; And

Operating a syringe pump of the capture recognition component immobilization device to draw a detection reagent present in the detection reagent processing device into the column to detect a combination of the capture recognition component and the analyte reacted in the matrix;

It relates to a method for analyzing biological samples for disease diagnosis.

EMBODIMENT OF THE INVENTION Hereinafter, this invention is demonstrated in detail.

In the present invention, the 'biological sample' means a sample material of all living organisms containing a labeling substance (analyte) for diagnosing a disease, for example, blood, plasma, serum, lymph, saliva, urine, tissue It may include crushed fluid, culture fluid, semen, sweat, bone marrow fluid, ocular fluid, brain extract, spinal fluid, joint fluid, thymus fluid, ascites, amniotic fluid and any other biological solution that contains other analytes. Preferably, blood, saliva, etc. can be mainly used.

In the present invention, the 'analyte' refers to a label included in a biological sample and to be diagnosed, for example, amino acids, proteins, peptides, enzymes, hormones, ligands, receptors, antigens, antibodies , Antibody fragments, cofactors, and the like.

In the present invention, the 'capture recognition component' means a substance capable of specifically recognizing and binding analytes, for example, amino acids, proteins, peptides, enzymes, hormones, ligands, receptors, antigens, antibodies, antibodies Fragments, cofactors, haptens, and the like.

For example, when the analyte is an antibody, the capture recognition component may be an antigenic protein recognized by the antibody. Conversely, when the analyte is a protein such as an antigen, the capture recognition component may selectively recognize the target antigen. Can be an antibody. As another example, when the analyte is a ligand, the capture recognition component may be a receptor protein specifically recognized by the ligand, and conversely, when the analyte is a receptor protein, the capture recognition component may be its ligand.

The present invention provides an apparatus for immobilizing a capture recognition component for analyzing a biological sample, by using a three-dimensional matrix structure in which the capture recognition component is immobilized, thereby maximizing the reaction participation of the biological sample, thereby improving sensitivity of diagnosis. .

The capture recognition component immobilization device of the present invention is specifically,

A column having both ends open and having a space therein;

A matrix present in the column and immobilized with capture components;

A porous membrane present in the column and layered on top and bottom of the matrix to which the capture recognition component is immobilized; And

And a syringe pump present in the column and at a distance from the matrix and the porous membrane.

The column providing the basic support of the capture recognition component immobilization device of the present invention is open at both ends and has a space therein, such that a matrix, a porous membrane and a syringe pump can be located therein, and one open end Through the sample, the washing solution or the detection reagent has a structure that can be inhaled or discharged into the interior space. The material of the column is not particularly limited, but may be manufactured using a transparent and easy-to-use material such as glass, high density polyethylene (HDPE) or ethylene terephthalate (PET).

Preferably, the open one end of the column forms a narrower structure, the end of which may form a needle-like passage, through which the sample, the washing solution, the detection reagent, or the like passes into the inner space. It is preferred to be inhaled or discharged.

The matrix to which the capture recognition component of the present invention is immobilized is a material capable of firmly fixing the capture recognition component by forming an irreversible covalent bond, and a porous matrix is preferable. For example, the polymer agarose, cellulose, glass beads, Natural matrices such as silica, alumina, zeolite, or synthetic matrices such as polyacrylamide beads, polymethacrylic beads, polystyrene beads, membranes, and the like can be commonly used.

The thickness of the matrix can be varied, for example from 0.1 μm to 100 cm or more, to suit the desired use of the device. The matrix can be used in the form of a single sheet or in stacked form to achieve the desired adsorption capacity.

The matrix is amine, thiol, carboxyl group, epoxy, formyl, tresyl, hydroxysuccinimide ester, sulfonic acid, primary amine, quaternary amine, carboxyl group, cyano, cyclohexyl, octyl and octadecyl groups, Reactive linkers, such as oxylane, N-carboxysuccinimide esters, sulfonyl esters, imidazole carbamates, are functionalized on the outer and / or outer surfaces, thereby trapping the capture components in an irreversible covalent bond to secure them .

The membrane or polymer matrix support used in the chip substrate has formed a two-dimensional structure, so that the fixation of the biomolecules is limited around the outer surface in contact with the outside, or when covalently bonded to increase the holding force of the biomolecules, There was a problem with the restriction of maintaining the biological activity of environmentally sensitive enzymes and other proteins. On the contrary, the porous matrix of the present invention has a high advantage because the utilization of the surface area is high and the various immobilization methods for maintaining the activity of the biomolecule can be utilized because the three-dimensional structure is made from the individual beads in which the biomolecules are already fixed. .

In the present invention, the porous membrane layered on the upper and lower ends of the matrix having the capture recognition component immobilized thereon is coated on the upper and lower ends of the matrix to prevent the outflow of the matrix, and forms a porous membrane inside the column and the matrix solution thereon. After putting the appropriate amount to build up sequentially from the bottom to cover the top of the porous membrane to prevent the outflow to the outside.

The porous membrane cannot pass through the matrix or the capture recognition component immobilized on the matrix, but passes through the sample, the washing solution or the detection reagent, etc., which enter through the terminal passage of the column, thereby trapping the capture recognition component and the sample, the washing solution or the like in the matrix. Allow detection reagents to react.

The syringe pump in the present invention includes any type of pump known in the art that can slide or rotate in the column, such as rotary pumps, piston pumps, and the like, to heat the sample and various solutions by pressurization or depressurization. In the lower part of the through the needle-like passage can be appropriately involved in the reaction.

For example, when the piston is withdrawn and the sample and various solutions are sucked into the closed air space in the column through the passage under the column, the sucked sample passes through the porous membrane and reaches the matrix in which the capture recognition component is immobilized. The reaction may occur, and after the reaction, the piston may be expanded to allow the sample or the like to pass through the porous membrane and exit the column.

The present invention also provides a method for producing a capture recognition component immobilization device for analyzing the biological sample, preferably the manufacturing method

Fixing the capture recognition component to the matrix;

Depositing a matrix in which the capture recognition component is immobilized in the column; And

Injecting a syringe pump into the column.

The first step is to fix the capture recognition component to the matrix, the usable matrix is a natural matrix such as polymer agarose, cellulose, glass beads, silica, alumina, zeolite or polyacrylamide beads, polymethacrylic acid beads It may be a porous matrix commonly used, such as synthetic matrices such as polystyrene beads, membranes, and the like, and may be amine, thiol, carboxyl group, epoxy, formyl, tresyl, hydroxysuccinimide ester, sulfonic acid, primary amine Reactive linkers such as quaternary amines, carboxyl groups, cyano, cyclohexyl, octyl and octadecyl groups, oxiranes, N-carboxysuccinimide esters, sulfonyl esters, imidazole carbamates, etc. Functionalized, the capture recognition component is tightly fixed by forming an irreversible covalent bond.

In the second step, a matrix in which the capture recognition component is immobilized is deposited in a column, and a porous membrane is first formed in a column made of a transparent, mass-produced material such as glass, HDPE, PET, etc. An appropriate amount of the matrix solution is put thereon to be sequentially stacked from the bottom, and then the top is covered with a porous membrane to prevent the outflow to the outside. In addition, there is a narrow open passage at the bottom of the column to allow the sample and various solutions to pass through and to participate in the reaction.

The third step is to put the syringe pump in the column, the syringe pump may use any type of pump known in the art, such as a rotary pump, a piston pump, it is present at a distance from the matrix and the porous membrane It is preferable.

The present invention also provides a biological sample analysis system for diagnosing diseases by using a capture recognition component immobilization device for analyzing a biological sample, thereby improving sensitivity of diagnosis by maximizing a reaction participation amount of the biological sample.

The biological sample analysis system of the present invention, specifically

The capture recognition component immobilization device;

A sample collection device;

Washing device; And

Detection reagent processing apparatus.

In the present invention, the capture recognition component immobilization device is as described above.

In the present invention, the sample collection device includes blood, plasma, serum, lymph, saliva, urine, tissue disruption fluid, culture fluid, semen, sweat, bone marrow fluid, ocular fluid, brain extract, spinal fluid, joint fluid, thymus fluid, ascites, amniotic fluid and A device containing a sample containing all other biological solutions containing other analytes.

In the present invention, the washing device is a device containing a salt or washing solution that is commonly used. Examples of the washing solution include phosphate buffered saline (PBS), Triton X100, bovine serum albumen (BSA), Tween 20, Tris, Tris / EDTA (TE), HEPES, TES, PIPES, and the like. The scope of the invention is not limited thereto.

In the present invention, the detection reagent processing apparatus includes a detection reagent that specifically recognizes a conjugate (eg, an antigen-antibody complex, a ligand-receptor complex, etc.) composed of a capture recognition component and an analyte, wherein the detection reagent is detected. Preferred antibodies are preferred. In the case of a detection antibody, a polyclonal antibody or a monoclonal antibody labeled with an enzyme capable of generating a detectable signal upon reaction with an appropriate substrate may be used. Examples of the enzyme include horseradish peroxidase. All enzymes commonly used, such as peroxidase enzymes, such as) or alkaline phosphatase, etc. can be used.

The biological sample analysis system of the present invention may include two or more washing devices and / or detection reagent processing devices according to an analysis method, and may automatically analyze the sample collection device, the cleaning device and the detection reagent processing device sequentially, or vice versa. Manual analysis can also be achieved by artificially adjusting the order of reactions.

Hereinafter, the biological sample analysis method using the analysis system will be described in more detail step by step.

The biological sample analysis method of the present invention,

Contacting one end of the capture recognition component immobilization device with the sample collection device;

Operating a syringe pump of the capture recognition component immobilization device to suck a sample present in the sample collection device into a column to react the capture recognition component in the matrix with the analyte in the sample;

Operating a syringe pump of the capture recognition component immobilization device to suck the washing solution present in the cleaning device into the column to wash components not involved in the reaction in the matrix;

And operating a syringe pump of the capture recognition component immobilization device to suck the detection reagent present in the detection reagent processing device into the column to detect a combination of the capture recognition component and the analyte reacted in the matrix.

The first step is a “sample contact step”, in which one open end of the capture recognition component immobilization device contacts the sample in the sample collection device so that the sample can be sucked into the column of the immobilization device. It may be by passiveization.

Samples available include blood, plasma, serum, lymph, saliva, urine, tissue lysate, culture fluid, semen, sweat, bone marrow fluid, eye fluid, brain extract, spinal fluid, joint fluid, thymus fluid, ascites, amniotic fluid and other analytes. As a sample containing all other biological solutions contained therein, preferably blood, saliva, and the like can be mainly used. Among them, blood contains various biomolecules and low-molecular substances in addition to the analyte, which is highly likely to cause background reactions with capture components and coexist with various cells. It is desirable to prepare a solution that can be diluted at least 5-fold containing a protease inhibitor so that it does not interfere with the binding between the capture recognition component and the analyte.

The second step is a “sample reaction step”, which operates a syringe pump of the capture recognition component immobilization device to suck a sample present in the sample collection device into a column to react the capture recognition component in the matrix with the analyte in the sample. Step.

The syringe pump is at the top at a distance from the matrix and the porous membrane in the column. For example, when using a piston pump, the sample in the sample device is forced through the passage at the end of the column by drawing the piston to reduce the internal pressure of the column. You can move it inside. Samples moved into the column pass through the porous membrane to the matrix and react with the capture components immobilized on the matrix, thereby forming specific binding between the analyte and the capture components in the sample (eg, antigen-antibody). Complexes, ligand-receptor complexes, etc.). Then, by expanding the piston to increase the internal pressure of the column, samples that do not participate in the reaction in the matrix can be passed through the porous membrane and then discharged into the column through the column ends.

The third step is a washing step, in which a syringe pump of the capture recognition component immobilization device is operated to suck the washing solution present in the cleaning device into the column, thereby washing the components that do not participate in the reaction in the matrix.

The washing step is a step for washing substances not participating in the reaction in the matrix. First, one end of the capture recognition element fixing device is contacted with the washing device, and then the piston is moved in the column in the same principle as in step 2 above. By varying the pressure of the cleaning solution, the cleaning solution can be used to clean materials that did not participate in the reaction in the matrix. After the wash is completed, the wash solution can be discharged out of the column via a piston movement.

The fourth step is a 'detection step', in which a syringe pump of the capture recognition component immobilization device is operated to draw a detection reagent present in the detection reagent processing device into a column, and a combination of the capture recognition component and the analyte reacted in the matrix. It is a step of detecting.

The detecting step is to detect the conjugate between the analyte and the capture recognition component in the matrix. First, one end of the capture recognition component immobilization device is contacted with the detection reagent processing device, and then the piston is operated in the same principle as in step 2 above. By changing the pressure inside the column by moving the detection reagent, the detection reagent can detect the conjugate between the analyte and the capture component in the matrix. After the reaction with the detection reagent is completed, the detection reagent solution may be discharged out of the column through the piston movement.

Furthermore, the analytical process can be appropriately modified by those skilled in the art. For example, an additional washing step and a second detection step may be further performed after the detection step, and in some cases, the steps may be reversed. In addition, the amount of analyte bound to the capture recognition component is quantitatively detected by reacting the detection substrate solution, and the presence or absence of the analyte is judged to be negative or positive by controlling the intensity of the signal displayed by controlling the detection sensitivity. It may further comprise a step.

The sample analysis system of the present invention uses a three-dimensional matrix structure in which a capture recognition component is immobilized to maximize the reaction participation rate of a biological sample for diagnosing a disease, thereby improving the detection limit by 10 to 100 times compared to the conventional ELISA method. As a result, the sensitivity of disease diagnosis can be improved and applied throughout the veterinary, medical, public health, environmental, food and biotechnology industries.

1 is a view showing a reaction vessel and a reaction method according to an embodiment of the present invention.
Description of the Related Art [0002]
1: syringe pump
2: column
3: porous membrane
4: Matrix in which the capture recognition component is immobilized
Figure 2 shows the result of detecting the rabbit anti-BP26 IgG of the target material according to the measuring method according to the present invention.

Hereinafter, the present invention will be described in detail by way of examples. However, the following examples are illustrative of the present invention, and the present invention is not limited by the following examples.

Example  1. Immobilization and Deposition of Capture Recognition Components

In the present invention, all commonly used protein immobilization methods may be used, and the method is not limited. The specific examples describe a method of immobilization using an aminolink resin immobilized by binding to an amine group of a capture recognition component. The capture recognition component used in the example is recombinant BP26 protein of Sobrucella.

First, 2 ml of aminolink resin (Thermo Corporation) was taken into a column, washed with 0.1 M sodium phosphate buffer, pH 7.0, and each of the capture recognition component and the NaCNBH3 solution was added and reacted at room temperature for 2 hours. After the filtrate was removed, 1M Tris-Cl, pH7.4 and NaCNBH3 solution were added to block unreacted linker and reacted at room temperature for 2 hours. After removal of the filtrate, all trace trace components that may be precipitated by 1M NaCl solution were removed, washed with PBS, and prepared.

The experiment was carried out as described below for the deposition of the matrix purified from the capture recognition component prepared by the proposed manufacturing method. The bottom of the vessel with a needle for deposition was closed with a porous membrane and 50ul ~ 200ul of the matrix was added. Solutions other than the matrix were removed by gravity and a porous membrane was placed again on top of the deposited matrix to prevent loss of the deposited matrix. Finally, the piston was fitted with the appropriate size of the vessel to prepare for subsequent reactions.

Example  2, analysis of analytes

The analyte used in this example is rabbit anti-BP26 IgG, which is an antibody against recombinant BP26 protein, a type of outer membrane protein of Sobru cell bacteria.

In addition, in order to demonstrate the superiority of the present embodiment, an experiment was performed to quantify and compare the amount of rabbit anti-BP26 IgG in a sample by using an ELISA technique using a multiwell microplate to which recombinant BP26 protein was attached.

Prior to performing the experiment, a plate to which recombinant BP26 protein was bound was prepared. To bind to each well of the plate, dilute the recombinant BP26 protein in a coating buffer (100 mM sodium bicarbonate / carbonate buffer, pH 9.6) at a concentration of 1 ug / ml and add 100 ul of each well to a maxisorp ELISA plate (Thermo corporation) at room temperature. After reaction for some time, remove the filtrate and wash with a washing buffer (PBS buffer containing 0.05% tween-20), and then add a blocking buffer (0.5% BSA, PBS buffer containing 0.05% tween-20) and room temperature After the reaction at 1 hour was discarded to prepare a plate for the reaction.

To perform the experiment, rabbit anti-BP26 IgG was prepared by diluting in FBS from 1ug / ml to 1pg / ml. For the use of this assay system, 1 ml of rabbit anti-BP26 IgG-diluted FBS is mixed with 4 ml of PBS solution containing 0.5% BSA and 0.05% tween-20 and diluted 1/5 in a container containing protein A. The piston was used to move up and down, and the reaction between samples occurred through 10 repeated piston movements. Thereafter, the solution was subjected to three piston movements for washing with PBS solution containing 0.05% tween-20. As an antibody for detection, goat anti-rabbit IgG solution in which HRP was bound was used, and the vertical movement was performed 10 times using a piston for binding. In order to remove the unbound detection antibody, the washing process was performed in the same manner as in the upper part, and 200 μl of the TMB solution serving as a substrate of the HRP enzyme was passed through the matrix by vertical movement using a piston, and the process was repeated 20 times. 200ul of 2N HCl solution was added to the solution to terminate the reaction, and the absorbance of OD450 was measured using an ELISA reader.

Meanwhile, the following experiment was conducted to quantify the amount of rabbit anti-BP26 IgG in a sample using an ELISA technique using a multiwell microplate to which recombinant BP26 protein is attached. First, 80ul of blocking buffer was first put into each well coated with recombinant BP26 protein, and 20ul of FBS solution in which rabbit anti-BP26 IgG was diluted was added, and reacted at room temperature for 1 hour and washed with the above washing solution. In this state, the goat anti-rabbit IgG solution in which HRP is bound is added, reacted, and washed again as above. After the reaction, the TMB solution, which is a substrate of HRP enzyme, is reacted for about 10 minutes, and then the reaction is stopped with 2N HCl. ELISA reader Absorbance of OD450 was measured using the instrument.

The results of the above experiment are shown in FIG. 2. As can be seen in the figure, the quantitation of rabbit anti-BP26 IgG using the conventional ELISA technique showed a sudden decrease in absorbance from 10 pg / ml, but 1 pg / ml reached a level almost impossible to quantify. In contrast, the present invention showed a high level of absorbance of about 1 pg / ml up to about 1 pg / ml, and fully shows the possibility of detecting up to 100 fg / ml by optimizing the solution composition and the number of reactions.

1: syringe pump
2: column
3: porous membrane
4: Matrix in which the capture recognition component is immobilized

Claims (11)

A column having both ends open and having a space therein;
A matrix present in the column and immobilized with capture components;
A porous membrane present in the column and in contact with the top and bottom of the matrix to which the capture recognition component is immobilized; And
A syringe pump present in the column and at a distance from the matrix and the porous membrane;
Capture recognition component immobilization device for the analysis of biological samples for disease diagnosis.
The disease of claim 1, wherein the capture recognition component is an amino acid, protein, peptide, enzyme, hormone, ligand, receptor, antigen, antibody, antibody fragment, cofactor or hapten capable of specifically binding to an analyte in a biological sample. Capture recognition component immobilization device for diagnostic biological sample analysis.
The method of claim 2, wherein the biological sample is blood, plasma, serum, lymph, saliva, urine, tissue disruption, culture, semen, sweat, bone marrow fluid, ocular fluid, brain extract, spinal fluid, joint fluid, thymus fluid, ascites and A capture recognition component immobilization device for analyzing a biological sample for disease diagnosis, which is a sample selected from the group consisting of amniotic fluid.
The method of claim 1, wherein the matrix is polymer agarose, cellulose, glass beads, silica, alumina, zeolite, polyacrylamide beads, polymethacrylic beads, polystyrene beads, or membranes. Capture recognition component immobilization device for the treatment.
The apparatus of claim 1, wherein the matrix is a porous matrix having a thickness of 0.1 μm to 100 cm.
The matrix of claim 1, wherein the matrix is an amine, thiol, carboxyl group, epoxy, formyl, tresyl, hydroxysuccinimide ester, sulfonic acid, carboxyl group, cyano, cyclohexyl, octyl and octadecyl group, oxylane, N A capture recognition component immobilization device for disease diagnosis biological sample, wherein the reactive linker selected from the group consisting of carboxysuccinimide ester, sulfonyl ester and imidazole carbamate is functionalized.
The apparatus of claim 1, wherein the column is made of glass, high density polyethylene (HDPE), or ethylene terephthalate (PET).
The apparatus of claim 1, wherein the syringe pump is a piston pump.
A capture recognition component immobilization device according to any one of claims 1 to 8;
A sample collection device;
Washing device; And
Comprising a detection reagent processing apparatus,
Biological sample analysis system for disease diagnosis.
Fixing the capture recognition component to the matrix;
Depositing a matrix in which the capture recognition component is immobilized in the column; And
Positioning a syringe pump in the column; Containing
A method for manufacturing a capture recognition component immobilization device for diagnosing a disease according to any one of claims 1 to 8.
Contacting one end of the capture recognition component immobilization device of claim 1 with a sample collection device;
Operating a syringe pump of the capture recognition component immobilization device to suck a sample present in the sample collection device into a column to react the capture recognition component in the matrix with the analyte in the sample;
Operating a syringe pump of the capture recognition component immobilization device to suck the washing solution present in the cleaning device into the column to wash components not involved in the reaction in the matrix; And
Operating a syringe pump of the capture recognition component immobilization device to draw a detection reagent present in the detection reagent processing device into the column to detect a combination of the capture recognition component and the analyte reacted in the matrix;
Methods of analyzing biological samples for disease diagnosis.

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