KR20120056442A - A microfluidic chip for analysis of biological fluid - Google Patents

Abstract

A microfluidic control chip is provided. The microfluidic control chip is formed by recessing a first portion of the upper surface of the substrate, and is formed by recessing a second portion of the upper surface of the substrate, the sample inlet portion into which the biological sample is injected, and a reaction sample reacting with the biological sample. A sample reaction portion provided by recessing the third portion of the upper surface of the substrate, a sample discharge portion for discharging the biological sample and the reaction sample, a first microfluidic channel connecting the sample inlet portion and the sample reaction portion, and a sample reaction portion; A second microfluidic channel connecting the sample outlet and a membrane attached to each of the first and second microfluidic channels to flow the sample.

Description

A microfluidic chip for analysis of biological fluid

The present invention relates to a microfluidic control chip for biological sample analysis, and more particularly to a microfluidic control chip having improved reactivity and analysis efficiency between a biological sample and a reaction sample.

Microfluidic control in the form of a lab-on-a-chip that integrates biochemical reactions such as bio-marker detection of a specific disease in the blood with minimal involvement by the experimenter Chips are being researched. In addition, researches for diagnosing diseases by analyzing specific proteins in blood or other body fluids have been actively conducted, and microfluidic control-based biochips for measuring and analyzing such specific proteins have been researched and developed. Furthermore, next-generation medical technologies are being developed that can register and view clinical information such as medical records, prescriptions, test results, and medication records in real time for immediate medical treatment on the spot. One of these is the point of care (POC) diagnosis. Technology is being researched.

On the other hand, the microfluidic control chip is in the form of a sandwich immune response using an antibody that can specifically bind to the protein and cells to be detected. In this method, microfluidic control is not easy, and antigen / antibody affinity is achieved. Problems such as an inefficient immune response occurs by the diagram has a problem of inferior accuracy and reproducibility.

The problem to be solved by the present invention is to provide a microfluidic control chip with improved reactivity and analysis efficiency between the biological sample and the reaction sample.

The problems to be solved by the present invention are not limited to the above-mentioned problems, and other problems not mentioned can be clearly understood by those skilled in the art from the following description.

In order to achieve the above object, the microfluidic control chip according to an embodiment of the present invention is formed by recessing a first portion of an upper surface of a substrate, and includes a sample inlet through which a biological sample is injected and an upper surface of the substrate. A sample reaction part is formed by recessing two portions and is provided by recessing a third portion of the upper surface of the substrate provided with a reaction sample that reacts with the biological sample, and a sample discharge part for discharging the biological sample and the reaction sample, and a sample inflow. And a first microfluidic channel connecting the sample and the sample reaction part, a second microfluidic channel connecting the sample reaction part and the sample outlet, and a membrane attached to each of the first and second microfluidic channels to flow the sample.

Specific details of other embodiments are included in the detailed description and the drawings.

According to a microfluidic control chip according to an embodiment of the present invention, microfluids are formed by forming grooves in a plate to form a sample inlet, a sample reaction part, and a sample outlet part, and connect the sample inlet part, the sample reaction part, and the sample outlet part. The sample can be smoothly flowed by attaching a membrane to the channels.

1 is a view showing a microfluidic control chip according to an embodiment of the present invention.

BRIEF DESCRIPTION OF THE DRAWINGS The advantages and features of the present invention, and how to accomplish them, will become apparent by reference to the embodiments described in detail below with reference to the accompanying drawings. However, the present invention is not limited to the embodiments disclosed below, but may be implemented in various forms. It is provided to fully convey the scope of the invention to those skilled in the art, and the present invention is defined only by the scope of the claims. Like reference numerals refer to like elements throughout the specification.

The terminology used herein is for the purpose of describing particular embodiments only and is not intended to be limiting of the invention. In the present specification, the singular form includes plural forms unless otherwise specified in the specification. As used herein, the terms 'comprises' and / or 'comprising' mean that the stated element, step, operation and / or element does not imply the presence of one or more other elements, steps, operations and / Or additions.

In addition, the embodiments described herein will be described with reference to cross-sectional and / or plan views, which are ideal exemplary views of the present invention. In the drawings, the thicknesses of films and regions are exaggerated for effective explanation of technical content. Accordingly, the regions illustrated in the figures have schematic attributes, and the shape of the regions illustrated in the figures is intended to illustrate a particular form of region of the device and not to limit the scope of the invention.

The microfluidic control chip according to an embodiment of the present invention may be used to detect proteins in blood, and more particularly, to analyze disease-related proteins and cells present in the blood of a patient such as cardiovascular disease or cancer. Can be.

The microfluidic control chip according to an embodiment of the present invention can be produced simply by making a groove on the plate, and can be applied to various lab-on-a-chip such as protein chips, microbiochemical analysis systems, microbiochemical reactors, and the like.

In addition, in the embodiments of the present invention, sandwich immuno-assay may be used to detect a specific protein.

In the microfluidic control chip according to an embodiment of the present invention, when detecting proteins or cells in the blood, grooves are formed in the plate to facilitate fluid control and increase the efficiency of the immune response. In addition, the grooves may be connected through the microfluidic channel, and the membrane may be attached to the microfluidic channel to facilitate fluid flow into the groove.

1 is a view showing a microfluidic control chip according to an embodiment of the present invention. Hereinafter, a microfluidic control chip according to an embodiment of the present invention will be described in detail with reference to FIG. 1.

Referring to FIG. 1, the microfluidic control chip 100 according to an embodiment may include lower and upper plates 110 and 150, a sample inlet 120, a sample reaction unit 130, and a sample discharger 140. And first and second microfluidic channels 125 and 135.

The microfluidic control chip 100 may be manufactured by combining the lower plate 110 and the upper plate 150. The lower and upper plates 110 and 150 may be formed of a transparent material to transmit light. For example, the lower and upper plates 110, 150 may be plastic, glass or quartz substrates. In addition, the lower and upper plates 110 and 150 have a high index of refraction such as titanium oxide (TiO ₂ ), tantalum oxide (Ta ₂ O ₅ ) or aluminum oxide (Al ₂ O ₃ ), SiN, or the like. Substrates can be used. In addition, the lower and upper plates 110 and 150 may include polydimethylsiloxane (PDMS), polymethylmethacrylate (PMMA), polycarbonate (PC), cyclic olefin copolymer (COC), polyamide (PA), polyethylene (PE), polypropylene (PP), Polyphenylene ether (PPE), polystyrene (PS), polyoxymethylene (POM), polyetheretherketone (PEEK), polytetrafluoroethylene (PTFE), polyvinylchloride (PVC), polyvinylidene fluoride (PVDF), polybutyleneterephthalate (PBT), fluorinated ethylenepropylene (PEP), and PFA (PFA) perfluoralkoxyalkane) and the like.

The lower plate 110 may include a sample inlet 120, a sample reaction unit 130, and a sample outlet 140. In detail, the sample inlet part 120, the sample reaction part 130, and the sample outlet part 140 may be formed by forming grooves in the lower plate 110, respectively.

The sample inlet unit 120, the sample reaction unit 130, and the sample outlet unit 140 may be formed by a photolithography process, an electron beam lithography process, or an imprint process of transferring a nano pattern using a stamp. Can be. In addition, the lower plate 110 having the grooves may be formed by injection molding. In an exemplary embodiment, three grooves are formed in the lower plate 110, but the number of grooves may be increased according to the number of proteins to be detected.

The sample inlet part 120 and the sample reaction part 130 are connected through the first microfluidic channel 125, and the sample reaction part 130 and the sample outlet part 140 connect the second microfluidic channel 135. Connected through. The first micro fluidic channel 125 moves the sample from the sample inlet 120 to the sample reaction unit 130. The second microfluidic channel 135 moves the sample from the sample reaction unit 130 to the sample discharge unit 140.

The first and second microfluidic channels 125 and 135 are formed by the lower and upper plates 110 and 150 spaced apart from each other by a predetermined interval. At this time, the spacing h between the lower and upper plates 110 and 150 is adjusted such that a capillarity force can be sufficiently acted. Accordingly, the sample may pass through the first and second microfluidic channels 125 and 135 by capillary force. In addition, the membranes 127 and 137 may be attached to the first and second microfluidic channels 125 and 135 to facilitate the flow of a fluid such as a biological sample. The membranes 127 and 137 may flow a sample from the sample inlet 120 or the sample reaction unit 130 to the sample reaction unit 130 or the sample discharge unit 140.

Membranes 127 and 137 may be used, for example, a nitrocellulose (NC) membrane, nylon or aligned nanotubes. In addition, the membranes 127 and 137 may be micro paper filters in which micropores are formed to pass only proteins that immunoreact with the probe materials of the sample reaction unit 130. The micro paper filter may vary in thickness and micropore size of the micro paper filter according to the amount of the body fluid introduced into the sample inlet 120 of the proteins included in the body fluid. For example, the membranes 127 and 137 may have a thickness of several tens to hundreds of micrometers and a pore size.

As the membranes 127 and 137 are attached to the first and second microfluidic channels 125 and 135, the sample may have a membrane (even if the capillary force is weak in the first and second microfluidic channels 125 and 135). 127, 137). In addition, the first and second microfluidic channels 125 and 135 may be hydrophilic surface treated so that the sample can be moved smoothly.

In the microfluidic control chip 100 according to an embodiment of the present invention, the sample injected into the sample inlet 110 may be a body fluid obtained from a living body. The body fluid may be, for example, blood, urine, saliva, and the like, and the body fluid may include not only the target substance to be detected, but also nonspecific molecules that do not bind to the probe substances. Specifically, the body fluid may include a large number of target substances, and in order to accurately and quickly detect a specific target substance to be diagnosed among the target substances, it is necessary to remove unnecessary target substances from the body fluid. For example, blood contains numerous blood cells and plasma and contains protein components such as fats, metabolites, moisture, enzymes, antigens, antibodies and various cells. And, the specific target substance to be detected is mainly present in plasma. That is, the target substance includes, for example, proteins, nucleic acids, viruses, cells, organic molecules and inorganic molecules, and in the case of protein molecules, any biomolecule such as antigens, antibodies, substrate proteins, enzymes, coenzymes, and the like can be used. And for nucleic acids, may be DNA, RNA, PNA, LNA or hybrids thereof.

The sample inlet 110 through which the sample is introduced may be provided with proteins labeled with a phosphor. In the sample inlet 120, a fluorescent material label for the antigen may be used as a labeling kit.

The sample reaction unit 130 immobilizes probe substances (eg, antibodies) that react with or bind to specific target substances (eg, antigens) of the disease or condition to be diagnosed. . The sensing substances may be proteins, cells, viruses, nucleic acids, organic molecules or inorganic molecules depending on the target substance to be detected. In the case of proteins, any target substance such as antigen, antibody, substrate protein, enzyme, coenzyme, It is possible. And for nucleic acids, may be DNA, RNA, PNA, LNA or hybrids thereof. In addition, the sensing materials may be fixed to the sample reaction unit 130 directly or using organic molecules as intermediate molecules. In addition, an active group may be induced in the sample reaction unit 130, for example, on the surface of the gold nanoparticles, a carboxyl group (-COOH), a thiol group (-SH), a hydroxyl group (-OH), a silane group, an amine Active groups such as groups or epoxy groups can be derived.

In addition, after the O ₂ plasma treatment, the sample reaction unit 130 may introduce an amine group into 3-Aminopropyltriethyoxysilane (APTES), and immobilize the antibody via N- (ε-Maleimidocaproyloxy) Sulfosuccinimide Ester (Sulfo-EMCS). have. Fixation of the fluorescent material label or antibody has been briefly described for the purpose of explanation, but the present invention is not limited thereto.

The sample discharge unit 140 may store the washing solution, and may be formed deeper and wider than the sample chamber 120 and the sample reaction unit 130.

After the sample analysis is completed in the microfluidic control chip 100, a washing solution such as PBS is supplied to supply the sample inlet 120, the sample reaction unit 130, the sample outlet 140, and the first and second fine particles. Clean the fluid channels 125, 135. After washing, the sample reaction unit 130 may measure the result of the immune response with a fluorescent signal. That is, when irradiating a sample containing biomaterials with incident light having an absorption wavelength of a fluorescent substance labeled with an antibody or antigen, a specific protein is detected using fluorescence generated from the fluorescent substance. Can be analyzed.

Although the embodiments of the present invention have been described above with reference to the accompanying drawings, those skilled in the art to which the present invention belongs may be embodied in other specific forms without changing the technical spirit or essential features of the present invention. You will understand that. It is therefore to be understood that the above-described embodiments are illustrative and not restrictive in every respect.

Claims (1)

A sample inlet formed by recessing a first portion of an upper surface of the substrate, into which a biological sample is injected;
A sample reaction part formed by recessing a second portion of the upper surface of the substrate and provided with a reaction sample reacting with the biological sample;
A sample discharge part formed by recessing a third portion of the upper surface of the substrate and discharging the biological sample and the reaction sample;
A first microfluidic channel connecting the sample inlet and the sample reaction part;
A second microfluidic channel connecting the sample reaction part and the sample discharge part; And
And a membrane attached to each of the first and second microfluidic channels to flow a sample.

Images